Invest Clin 65(3): 321 - 334, 2024 https://doi.org/10.54817/IC.v65n3a05
Corresponding Author: Halis Suleyman, Erzincan Binali Yildirim University, Faculty of Medicine, Department of Phar-
macology, Erzincan, Turkey, Phone: +90 530 9211909, Fax: +90 446 2261819. E-mail: halis.suleyman@gmail.com
Protective effects of thiamine pyrophosphate
and cinnamon against oxidative liver damage
induced by an isoniazid and rifampicin
combination in rats.
Bahtınur Yeter1, Renad Mammadov2, Zeynep Koc3, Seval Bulut2, Tugba Bal Tastan4,
Mine Gulaboglu5 and Halis Suleyman2
1
Department of Child Health and Diseases, Faculty of Medicine, Erzincan Binali Yildirim
University, Erzincan, Turkey.
2
Department of Pharmacology, Faculty of Medicine, Erzincan Binali Yildirim University,
Erzincan, Turkey.
3Department of Biochemistry, Faculty of Medicine, Erzincan Binali Yıldırım University,
Turkey.
4
Department of Histology and Embryology, Faculty of Medicine, Erzincan Binali Yildirim
University, Erzincan, Turkey.
5
Department of Biochemistry, Faculty of Pharmacy, Ataturk University, Turkey.
Keywords: isoniazid; rifampicin; thiamine pyrophosphate; cinnamon extract; oxidative
stress; inflammation.
Abstract. Isoniazid and rifampicin (IRC) have been shown to cause hepa-
totoxicity in both clinical and preclinical studies. Oxidative stress and in-
flammation have been held responsible for the pathogenesis of IRC-induced
hepatotoxicity. Antioxidative and anti-inflammatory effects of thiamine py-
rophosphate (TPP) and cinnamon extract (CE) have been shown in previous
studies. Therefore, our study investigated the protective effects of TPP and CE
on possible liver damage caused by IRC treatment in rats. Twenty-four albino
Wistar rats were categorized into four groups: a healthy group (HG), an IRC
group (IRG), a TPP+IRC group (TIRG), and a CE+IRC group (CIRG). TPP (25
mg/kg) was administered intraperitoneally to TIRG, while CE (100 mg/kg)
was administered orally to CIRG. In IRG, TIRG, and CIRG, isoniazid (50 mg/
kg) and rifampicin (50 mg/kg) were administered orally one hour after these
treatments. For seven days, this procedure was repeated once a day. After this
period, blood samples were taken from the tail veins, and the rats were sac-
rificed. The removed liver tissues were analyzed for oxidant, antioxidant, and
proinflammatory cytokines and subjected to histopathological evaluation.
Serum alanine aminotransferase and aspartate aminotransferase activities
Corresponding Author: Halis Suleyman, Erzincan Binali Yildirim University, Faculty of Medicine, Department of Phar-
macology, Erzincan, Turkey, Phone: +90 530 9211909, Fax: +90 446 2261819. E-mail: halis.suleyman@gmail.com
Protective effects of thiamine pyrophosphate
and cinnamon against oxidative liver damage
induced by an isoniazid and rifampicin
combination in rats.
Bahtınur Yeter1, Renad Mammadov2, Zeynep Koc3, Seval Bulut2, Tugba Bal Tastan4,
Mine Gulaboglu5 and Halis Suleyman2
1
Department of Child Health and Diseases, Faculty of Medicine, Erzincan Binali Yildirim
University, Erzincan, Turkey.
2
Department of Pharmacology, Faculty of Medicine, Erzincan Binali Yildirim University,
Erzincan, Turkey.
3Department of Biochemistry, Faculty of Medicine, Erzincan Binali Yıldırım University,
Turkey.
4
Department of Histology and Embryology, Faculty of Medicine, Erzincan Binali Yildirim
University, Erzincan, Turkey.
5
Department of Biochemistry, Faculty of Pharmacy, Ataturk University, Turkey.
Keywords: isoniazid; rifampicin; thiamine pyrophosphate; cinnamon extract; oxidative
stress; inflammation.
Abstract. Isoniazid and rifampicin (IRC) have been shown to cause hepa-
totoxicity in both clinical and preclinical studies. Oxidative stress and in-
flammation have been held responsible for the pathogenesis of IRC-induced
hepatotoxicity. Antioxidative and anti-inflammatory effects of thiamine py-
rophosphate (TPP) and cinnamon extract (CE) have been shown in previous
studies. Therefore, our study investigated the protective effects of TPP and CE
on possible liver damage caused by IRC treatment in rats. Twenty-four albino
Wistar rats were categorized into four groups: a healthy group (HG), an IRC
group (IRG), a TPP+IRC group (TIRG), and a CE+IRC group (CIRG). TPP (25
mg/kg) was administered intraperitoneally to TIRG, while CE (100 mg/kg)
was administered orally to CIRG. In IRG, TIRG, and CIRG, isoniazid (50 mg/
kg) and rifampicin (50 mg/kg) were administered orally one hour after these
treatments. For seven days, this procedure was repeated once a day. After this
period, blood samples were taken from the tail veins, and the rats were sac-
rificed. The removed liver tissues were analyzed for oxidant, antioxidant, and
proinflammatory cytokines and subjected to histopathological evaluation.
Serum alanine aminotransferase and aspartate aminotransferase activities
322 Yeter et al.Investigación Clínica 65(3): 2024
Efectos protectores del pirofosfato de tiamina y la canela
contra el daño hepático oxidativo inducido por la combinación
de isoniazida y rifampicina en ratas.
Invest Clin 2024; 65 (3): 321 – 334
Palabras clave: isoniazida; rifampicina; pirofosfato de tiamina; extracto de canela; estrés
oxidativo; inflamación.
Resumen. Se ha demostrado que la isoniazida y la rifampicina (IRC) cau-
san hepatotoxicidad tanto en estudios clínicos como preclínicos. El estrés
oxidativo y la inflamación se han considerado responsables de la patogénesis
de la hepatotoxicidad inducida por IRC. En estudios anteriores se han demos-
trado los efectos antioxidantes y antiinflamatorios del pirofosfato de tiamina
(TPP) y del extracto de canela (CE). Por lo tanto, en nuestro estudio se inves-
tigaron los efectos protectores del TPP y el CE sobre el posible daño hepático
causado por el tratamiento con IRC en ratas. Se clasificaron 24 ratas Wistar
albinas en cuatro grupos: un grupo sano (HG), un grupo IRC (IRG), un grupo
TPP+IRC (TIRG) y un grupo CE+IRC (CIRG). El TPP (25 mg/kg) se admi-
nistró por vía intraperitoneal al TIRG, mientras que el CE (100 mg/kg) se ad-
ministró por vía oral al CIRG. En IRG, TIRG y CIRG, se administró isoniazida
(50 mg/kg) y rifampicina (50 mg/kg) por vía oral una hora después de estos
tratamientos. Durante siete días, este procedimiento se repitió una vez al día.
Tras este periodo, se tomaron muestras de sangre de las venas de la cola y se
sacrificaron las ratas. Los tejidos hepáticos extraídos se analizaron en busca
de citocinas oxidantes, antioxidantes y proinflamatorias y se sometieron a una
evaluación histopatológica. También se midieron las actividades séricas de la
alanina aminotransferasa y la aspartato aminotransferasa. Con el tratamien-
to con IRC se observó un aumento de los niveles de malondialdehído, factor
nuclear kappa B, factor de necrosis tumoral alfa, interleucina 1 beta e inter-
leucina 6, una disminución de los niveles totales de glutatión y de las activi-
dades de superóxido dismutasa y catalasa, y un aumento de las actividades de
alanina aminotransferasa y aspartato aminotransferasa (p<0,001). El análisis
histopatológico del IRG sugirió hepatotoxicidad (p<0,001). El TPP y el CE
were also measured. An increase in malondialdehyde, nuclear factor kappa
B, tumor necrosis factor-alpha, interleukin 1 beta, and interleukin-6 levels, a
decrease in total glutathione levels, superoxide dismutase and catalase activi-
ties, and an increase in alanine aminotransferase and aspartate aminotrans-
ferase activities were found with IRC treatment (p<0.001). The histopatho-
logical analysis of the IRG suggested hepatotoxicity (p<0.001). TPP and CE
administered with IRC inhibited the biochemical changes (p<0.001). In the
TIRG, this inhibition was higher than in the CIRG (p<0.05). Histological
damage was inhibited by TPP (p<0.001). CE prevented biochemical changes
but not histological changes except inflammatory cell infiltration. Therefore,
TPP may be better than CE in preventing IRC-induced hepatotoxicity.
Efectos protectores del pirofosfato de tiamina y la canela
contra el daño hepático oxidativo inducido por la combinación
de isoniazida y rifampicina en ratas.
Invest Clin 2024; 65 (3): 321 – 334
Palabras clave: isoniazida; rifampicina; pirofosfato de tiamina; extracto de canela; estrés
oxidativo; inflamación.
Resumen. Se ha demostrado que la isoniazida y la rifampicina (IRC) cau-
san hepatotoxicidad tanto en estudios clínicos como preclínicos. El estrés
oxidativo y la inflamación se han considerado responsables de la patogénesis
de la hepatotoxicidad inducida por IRC. En estudios anteriores se han demos-
trado los efectos antioxidantes y antiinflamatorios del pirofosfato de tiamina
(TPP) y del extracto de canela (CE). Por lo tanto, en nuestro estudio se inves-
tigaron los efectos protectores del TPP y el CE sobre el posible daño hepático
causado por el tratamiento con IRC en ratas. Se clasificaron 24 ratas Wistar
albinas en cuatro grupos: un grupo sano (HG), un grupo IRC (IRG), un grupo
TPP+IRC (TIRG) y un grupo CE+IRC (CIRG). El TPP (25 mg/kg) se admi-
nistró por vía intraperitoneal al TIRG, mientras que el CE (100 mg/kg) se ad-
ministró por vía oral al CIRG. En IRG, TIRG y CIRG, se administró isoniazida
(50 mg/kg) y rifampicina (50 mg/kg) por vía oral una hora después de estos
tratamientos. Durante siete días, este procedimiento se repitió una vez al día.
Tras este periodo, se tomaron muestras de sangre de las venas de la cola y se
sacrificaron las ratas. Los tejidos hepáticos extraídos se analizaron en busca
de citocinas oxidantes, antioxidantes y proinflamatorias y se sometieron a una
evaluación histopatológica. También se midieron las actividades séricas de la
alanina aminotransferasa y la aspartato aminotransferasa. Con el tratamien-
to con IRC se observó un aumento de los niveles de malondialdehído, factor
nuclear kappa B, factor de necrosis tumoral alfa, interleucina 1 beta e inter-
leucina 6, una disminución de los niveles totales de glutatión y de las activi-
dades de superóxido dismutasa y catalasa, y un aumento de las actividades de
alanina aminotransferasa y aspartato aminotransferasa (p<0,001). El análisis
histopatológico del IRG sugirió hepatotoxicidad (p<0,001). El TPP y el CE
were also measured. An increase in malondialdehyde, nuclear factor kappa
B, tumor necrosis factor-alpha, interleukin 1 beta, and interleukin-6 levels, a
decrease in total glutathione levels, superoxide dismutase and catalase activi-
ties, and an increase in alanine aminotransferase and aspartate aminotrans-
ferase activities were found with IRC treatment (p<0.001). The histopatho-
logical analysis of the IRG suggested hepatotoxicity (p<0.001). TPP and CE
administered with IRC inhibited the biochemical changes (p<0.001). In the
TIRG, this inhibition was higher than in the CIRG (p<0.05). Histological
damage was inhibited by TPP (p<0.001). CE prevented biochemical changes
but not histological changes except inflammatory cell infiltration. Therefore,
TPP may be better than CE in preventing IRC-induced hepatotoxicity.
Thiamine pyrophosphate and cinnamon protect rats from induced liver damage 323Vol. 65(3): 321 - 334, 2024
INTRODUCTİON
Isoniazid and rifampicin are anti-my-
cobacterial drugs that are part of the World
Health Organisation’s recommended com-
bination treatment regimen for tuberculo-
sis1
. Treatment for tuberculosis consists of
a quartet of anti-mycobacterial medications,
including isoniazid, rifampicin, ethambutol,
and pyrazinamide. Furthermore, the isonia-
zid and rifampicin combination (IRC) in-
cludes the supportive phase of tuberculosis
treatment 1,2 . Although effective in treating
tuberculosis, these drugs may cause serious
side effects. During treatment, there may be
toxic effects on the dermatology, gastroin-
testinal system, hypersensitivity, neurology,
hematology, and renal system 3
. Antituber-
culosis drugs can cause liver toxicity, which
can result in the discontinuation of the drug
as well as morbidity and mortality 4
. A pri-
mary antituberculosis drug, isoniazid, has
been reported to cause liver injury through
oxidative stress 5
. The role of lipid peroxida-
tion (LPO) in the pathogenesis of isoniazid-
associated oxidative liver injury has been
documented 6
. Severe oxidative liver damage
is believed to be initiated by reactive oxygen
species (ROS) 7
. According to the literature,
isoniazid has hepatotoxic effects due to the
formation of the acetylhydrazine metabolite
due to its acetylation 1
. Although rifampicin
alone has a low hepatotoxicity potential 6
, it
exhibits additive and synergistic hepatotox-
icity when used with isoniazid in treating
tuberculosis 4
. This has been attributed to
the stimulation of hydrolases by rifampicin
and the formation of hepatotoxic reactive
metabolites from isoniazid 8
.
The protective effect of thiamine pyro-
phosphate (TPP), thiamine’s active metabo-
lite, was evaluated in IRC-induced hepatotox-
icity in rats 9
. Evidence in the literature shows
that TPP protects various organs from oxida-
tive and proinflammatory cytokine damage
10-12
. Cinnamon, which we also investigate in
our study against the possible toxicity of IRC
on the liver, is the bark of some Cinnamo-
mum (Lauraceae) species 13
. Cinnamon has
been shown to have anti-inflammatory, anti-
oxidant, and many other beneficial biologi-
cal properties in the literature 14
. It has been
reported that cinnamon protects the heart,
liver, kidney, blood, brain, and spleen from
the toxicity of chemicals through its anti-
oxidant, radical-scavenging, and LPO-sup-
pressing properties15
. The anti-inflammatory
effect of cinnamon extract (CE) has been as-
sociated with inhibition of proinflammatory
cytokines 16
. This information suggests that
TPP and CE may be helpful in treating IRC-
induced liver injury. This study was designed
to investigate the protective effects of TPP
and CE against IRC-induced liver injury in
rats.
MATERİALS AND METHODS
Animals
Twenty-four male albino Wistar rats
(282-294 g, 5-6 months old) purchased from
the Experimental Animal Research and Ap-
administrados con el IRC inhibieron los cambios bioquímicos (p<0,001). En
el TIRG, esta inhibición fue mayor que en el CIRG (p<0,05). El daño histoló-
gico fue inhibido por el TPP (p<0,001). El CE previno los cambios bioquími-
cos pero no los histológicos, excepto la infiltración de células inflamatorias.
Por lo tanto, TPP puede ser una mejor opción que CE para la prevención de la
hepatotoxicidad inducida por IRC.
Received: 24-01-2024 Accepted: 28-04-2024
INTRODUCTİON
Isoniazid and rifampicin are anti-my-
cobacterial drugs that are part of the World
Health Organisation’s recommended com-
bination treatment regimen for tuberculo-
sis1
. Treatment for tuberculosis consists of
a quartet of anti-mycobacterial medications,
including isoniazid, rifampicin, ethambutol,
and pyrazinamide. Furthermore, the isonia-
zid and rifampicin combination (IRC) in-
cludes the supportive phase of tuberculosis
treatment 1,2 . Although effective in treating
tuberculosis, these drugs may cause serious
side effects. During treatment, there may be
toxic effects on the dermatology, gastroin-
testinal system, hypersensitivity, neurology,
hematology, and renal system 3
. Antituber-
culosis drugs can cause liver toxicity, which
can result in the discontinuation of the drug
as well as morbidity and mortality 4
. A pri-
mary antituberculosis drug, isoniazid, has
been reported to cause liver injury through
oxidative stress 5
. The role of lipid peroxida-
tion (LPO) in the pathogenesis of isoniazid-
associated oxidative liver injury has been
documented 6
. Severe oxidative liver damage
is believed to be initiated by reactive oxygen
species (ROS) 7
. According to the literature,
isoniazid has hepatotoxic effects due to the
formation of the acetylhydrazine metabolite
due to its acetylation 1
. Although rifampicin
alone has a low hepatotoxicity potential 6
, it
exhibits additive and synergistic hepatotox-
icity when used with isoniazid in treating
tuberculosis 4
. This has been attributed to
the stimulation of hydrolases by rifampicin
and the formation of hepatotoxic reactive
metabolites from isoniazid 8
.
The protective effect of thiamine pyro-
phosphate (TPP), thiamine’s active metabo-
lite, was evaluated in IRC-induced hepatotox-
icity in rats 9
. Evidence in the literature shows
that TPP protects various organs from oxida-
tive and proinflammatory cytokine damage
10-12
. Cinnamon, which we also investigate in
our study against the possible toxicity of IRC
on the liver, is the bark of some Cinnamo-
mum (Lauraceae) species 13
. Cinnamon has
been shown to have anti-inflammatory, anti-
oxidant, and many other beneficial biologi-
cal properties in the literature 14
. It has been
reported that cinnamon protects the heart,
liver, kidney, blood, brain, and spleen from
the toxicity of chemicals through its anti-
oxidant, radical-scavenging, and LPO-sup-
pressing properties15
. The anti-inflammatory
effect of cinnamon extract (CE) has been as-
sociated with inhibition of proinflammatory
cytokines 16
. This information suggests that
TPP and CE may be helpful in treating IRC-
induced liver injury. This study was designed
to investigate the protective effects of TPP
and CE against IRC-induced liver injury in
rats.
MATERİALS AND METHODS
Animals
Twenty-four male albino Wistar rats
(282-294 g, 5-6 months old) purchased from
the Experimental Animal Research and Ap-
administrados con el IRC inhibieron los cambios bioquímicos (p<0,001). En
el TIRG, esta inhibición fue mayor que en el CIRG (p<0,05). El daño histoló-
gico fue inhibido por el TPP (p<0,001). El CE previno los cambios bioquími-
cos pero no los histológicos, excepto la infiltración de células inflamatorias.
Por lo tanto, TPP puede ser una mejor opción que CE para la prevención de la
hepatotoxicidad inducida por IRC.
Received: 24-01-2024 Accepted: 28-04-2024
324 Yeter et al.Investigación Clínica 65(3): 2024
plication Centre of Erzincan Binali Yildirim
University were used in the study. The rats
were housed in an environment with ap-
propriate temperature (22±2°C), humidity
(50-60%) and 12-h light-dark cycle and fed
ad libitum. Experimental procedures were
carried out after the approval of the local
Animal Experiments Ethics Committee of
Erzincan Binali Yıldırım University (Date:
31.08.2023, Decision number: 29).
Chemical substances
Isoniazid and Rifampicin were procured
from Koçak Farma Ilaç ve Kimya Sanayi (Is-
tanbul, Turkey), Thiamine pyrophosphate
was procured from BioPharma (Moscow,
Russia), Cinnamon extract was procured
from Solgar (Leonia, USA), and thiopental
sodium used for the experiment was pro-
cured from IE Ulagay (Istanbul, Turkey).
Experimental groups
The rats (n=6/each group) were ran-
domly divided into healthy control (HG), iso-
niazid + rifampicin (IRG), TPP + isoniazid
+ rifampicin (TIRG), and CE + isoniazid +
rifampicin (CIRG) groups.
Experimental process
In the experiment, TPP (25 mg/kg) was
administered intraperitoneally to the TIRG.
CE (100 mg/kg) was administered orally
by gavage to the CIRG. In both the HG and
IRG, distilled water was administered the
same way. To the IRG, TIRG, and CIRG, iso-
niazid (50 mg/kg) and rifampicin (50 mg/
kg) were given orally one hour after the TPP,
CE, and distilled water were given. The indi-
cated treatment protocol was administered
once daily for seven days. At the end of the
seventh day, blood samples were collected
from the tail vein of the rats for alanine
aminotransferase (ALT) and aspartate ami-
notransferase (AST) analyses. The rats were
then euthanized with 50 mg/kg thiopental
sodium intraperitoneally, and liver tissues
were collected. Malondialdehyde (MDA),
total glutathione (tGSH), superoxide dis-
mutase (SOD), catalase (CAT), nuclear fac-
tor kappa B (NF-κB), tumor necrosis factor-
alpha (TNF-α), interleukin 1 beta (IL-1β),
and interleukin 6 (IL-6) were analyzed in
the liver tissues. Liver tissues were also ex-
amined histopathologically.
Biochemical analyses
Preparation of the samples
The tissues were cleaned of blood using
physiological saline and pulverized by adding
liquid nitrogen. Powdered tissues were dis-
solved in 50 mM phosphate buffer (pH=7.4).
For MDA, tGSH, SOD, CAT, NF-κB, TNF-α, IL-
1β, and IL-6 analyses, the supernatants ob-
tained after centrifugation were used.
Tissue MDA, tGSH, SOD, and CAT
determination
MDA and tGSH levels and SOD activities
in tissues were determined by measuring each
with Enzyme-Linked ImmunoSorbent Assay
(ELISA) kit (product nos. 10009055, 703002,
and 706002, respectively, Cayman Chemical
Company) according to the instructions. The
CAT analysis was conducted according to the
method recommended by Goth 17
.
Tissue NF-κB, TNF-α, IL-1β, and IL-6
determination
TNF-α, IL-1β, and IL-6 levels were deter-
mined with ELISA kits purchased from East-
biopharm Co Ltd (China), and NF-κB levels
were measured with commercial ELISA kits
purchased from SunRed Biological Technol-
ogy Co. Ltd (China). The analyses were per-
formed according to the kit instructions.
Determination of ALT and AST in
serum. Tubes without anticoagulants were
used for blood samples, and serum was used
for analyses. After centrifugation, the clear
filtrate was separated and stored at -80°C.
ALT and AST activities were determined
spectrophotometrically using a Cobas 8000
autoanalyzer (Roche Diagnostics GmBH,
Germany) with kits (Roche Diagnostics).
plication Centre of Erzincan Binali Yildirim
University were used in the study. The rats
were housed in an environment with ap-
propriate temperature (22±2°C), humidity
(50-60%) and 12-h light-dark cycle and fed
ad libitum. Experimental procedures were
carried out after the approval of the local
Animal Experiments Ethics Committee of
Erzincan Binali Yıldırım University (Date:
31.08.2023, Decision number: 29).
Chemical substances
Isoniazid and Rifampicin were procured
from Koçak Farma Ilaç ve Kimya Sanayi (Is-
tanbul, Turkey), Thiamine pyrophosphate
was procured from BioPharma (Moscow,
Russia), Cinnamon extract was procured
from Solgar (Leonia, USA), and thiopental
sodium used for the experiment was pro-
cured from IE Ulagay (Istanbul, Turkey).
Experimental groups
The rats (n=6/each group) were ran-
domly divided into healthy control (HG), iso-
niazid + rifampicin (IRG), TPP + isoniazid
+ rifampicin (TIRG), and CE + isoniazid +
rifampicin (CIRG) groups.
Experimental process
In the experiment, TPP (25 mg/kg) was
administered intraperitoneally to the TIRG.
CE (100 mg/kg) was administered orally
by gavage to the CIRG. In both the HG and
IRG, distilled water was administered the
same way. To the IRG, TIRG, and CIRG, iso-
niazid (50 mg/kg) and rifampicin (50 mg/
kg) were given orally one hour after the TPP,
CE, and distilled water were given. The indi-
cated treatment protocol was administered
once daily for seven days. At the end of the
seventh day, blood samples were collected
from the tail vein of the rats for alanine
aminotransferase (ALT) and aspartate ami-
notransferase (AST) analyses. The rats were
then euthanized with 50 mg/kg thiopental
sodium intraperitoneally, and liver tissues
were collected. Malondialdehyde (MDA),
total glutathione (tGSH), superoxide dis-
mutase (SOD), catalase (CAT), nuclear fac-
tor kappa B (NF-κB), tumor necrosis factor-
alpha (TNF-α), interleukin 1 beta (IL-1β),
and interleukin 6 (IL-6) were analyzed in
the liver tissues. Liver tissues were also ex-
amined histopathologically.
Biochemical analyses
Preparation of the samples
The tissues were cleaned of blood using
physiological saline and pulverized by adding
liquid nitrogen. Powdered tissues were dis-
solved in 50 mM phosphate buffer (pH=7.4).
For MDA, tGSH, SOD, CAT, NF-κB, TNF-α, IL-
1β, and IL-6 analyses, the supernatants ob-
tained after centrifugation were used.
Tissue MDA, tGSH, SOD, and CAT
determination
MDA and tGSH levels and SOD activities
in tissues were determined by measuring each
with Enzyme-Linked ImmunoSorbent Assay
(ELISA) kit (product nos. 10009055, 703002,
and 706002, respectively, Cayman Chemical
Company) according to the instructions. The
CAT analysis was conducted according to the
method recommended by Goth 17
.
Tissue NF-κB, TNF-α, IL-1β, and IL-6
determination
TNF-α, IL-1β, and IL-6 levels were deter-
mined with ELISA kits purchased from East-
biopharm Co Ltd (China), and NF-κB levels
were measured with commercial ELISA kits
purchased from SunRed Biological Technol-
ogy Co. Ltd (China). The analyses were per-
formed according to the kit instructions.
Determination of ALT and AST in
serum. Tubes without anticoagulants were
used for blood samples, and serum was used
for analyses. After centrifugation, the clear
filtrate was separated and stored at -80°C.
ALT and AST activities were determined
spectrophotometrically using a Cobas 8000
autoanalyzer (Roche Diagnostics GmBH,
Germany) with kits (Roche Diagnostics).
Thiamine pyrophosphate and cinnamon protect rats from induced liver damage 325Vol. 65(3): 321 - 334, 2024
Histopathological analysis
Tissue samples were fixed in formalin
(10%). Subsequently, tissue samples were
washed. Tissues were embedded in paraffin
after ethanol (70-100%) and xylol proce-
dures. Sections (4-5 μ) were prepared and
stained with hematoxylin-eosin (H&E). Sec-
tions were photographed and analyzed using
a light microscope (Olympus Inc., Tokyo,
Japan) and the DP2-SAL firmware program.
Histopathological changes in liver tissue
were defined as hepatocyte degeneration,
Kupffer cell activation, capillary congestion,
and the presence of polymorphonuclear cell
(PMNL) infiltration. Each section was grad-
ed 0-3 for each criterion (0, absent; 1, mild;
2, moderate; 3, severe). The evaluation was
performed by a pathologist who was unaware
of the study groups’ assignments.
Statistical analyses
The statistical procedures were con-
ducted using “SPSS for Windows, 22.0” sta-
tistical software. It was determined that the
numerical data were normally distributed
by the Shapiro-Wilk test; therefore, one-way
ANOVA was used for the analysis. Accord-
ing to the results of Levene’s test, Tukey
HSD or Games Howell tests were used as
post hoc tests for intergroup comparisons.
The biochemical data were presented as
mean ± standard deviation (X±SD). The
Kruskal-Wallis test was preferred for analyz-
ing semiquantitative histopathologic grad-
ing data, and then the post-hoc Dunn’s test
was employed for the analysis. Statistical
results were presented as median (quartile
1- quartile 3). The significance level was set
at p<0.05.
RESULTS
Analysis of MDA, tGSH, SOD and CAT
levels in liver tissue
It can be seen from Fig. 1A that MDA
levels in liver tissue increased in the IRG
group (5.48±0.20) compared to healthy
rats (2.39±0.18) (p<0.001). The increase
in MDA was observed to be suppressed in the
TIRG (2.67±0.24) and CIRG (4.06±0.15)
(p<0.001). Compared with CE, TPP sup-
pressed the increase in MDA more effectively
(p<0.001). The MDA levels in TIRG and HG
were not significantly different (p=0.100).
In IRG, a decrease in tGSH levels
and SOD and CAT activities (3.42±0.24,
5.33±0.15, 3.78±0.12, respectively) were
detected with the increase in MDA compared
to HG (6.34±0.18, 9.19±0.12, 7.51±0.21,
respectively) (p<0.001). TPP (5.80±0.10,
8.76±0.13, 7.09±0.26, respectively) and CE
(4.48±0.09, 6.66±0.16, 5.39±0.37, respec-
tively) inhibited the decrease in tGSH levels
and SOD and CAT activities (p<0.001). TPP
treatment prevented the decrease in antioxi-
dants more effectively than CE (p<0.001)
(Fig. 1B-D).
Analysis of NF-κB, TNF-α, IL-1β and IL-6
levels in liver tissue
As presented in Fig. 2A-D, the levels of
NF-κB, TNF-α, IL-1β, and IL-6 in the liver tis-
sues of rats in the IRG group (4.86±0.09,
4.68±0.09, 6.29±0.15, 5.70±0.12, respec-
tively) were higher than those in the HG
group (2.38±0.11, 2.13±0.07, 3.37±0.19,
2.49±0.24, respectively) (p<0.001). Com-
pared to the IRG group, both TPP (2.57±0.12,
2.37±0.11, 3.68±0.26, 2.77±0.11, respec-
tively) and CE (3.25±0.15, 3.31±0.14,
5.00±0.48, 4.02±0.22, respectively) inhib-
ited these increases in NF-κB, TNF-α, IL-1β
and IL-6 (p<0.05). However, this inhibition
was more significant in the TPP than in the
CE group (p<0.05). There were similari-
ties between TIRG and HG in terms of NF-
κB (p=0.315), IL-1β (p=0.157) and IL-6
(p=0.064) levels.
Analysis of ALT and AST activities
in serum samples
As presented in Fig. 3A-B, ALT and AST
activities measured from the serum of rats in
the IRG (86.67±4.55, 220.00±7.72, respec-
tively) were found to be increased according
to HG (29.17±4.02, 39.17±4.92, respec-
Histopathological analysis
Tissue samples were fixed in formalin
(10%). Subsequently, tissue samples were
washed. Tissues were embedded in paraffin
after ethanol (70-100%) and xylol proce-
dures. Sections (4-5 μ) were prepared and
stained with hematoxylin-eosin (H&E). Sec-
tions were photographed and analyzed using
a light microscope (Olympus Inc., Tokyo,
Japan) and the DP2-SAL firmware program.
Histopathological changes in liver tissue
were defined as hepatocyte degeneration,
Kupffer cell activation, capillary congestion,
and the presence of polymorphonuclear cell
(PMNL) infiltration. Each section was grad-
ed 0-3 for each criterion (0, absent; 1, mild;
2, moderate; 3, severe). The evaluation was
performed by a pathologist who was unaware
of the study groups’ assignments.
Statistical analyses
The statistical procedures were con-
ducted using “SPSS for Windows, 22.0” sta-
tistical software. It was determined that the
numerical data were normally distributed
by the Shapiro-Wilk test; therefore, one-way
ANOVA was used for the analysis. Accord-
ing to the results of Levene’s test, Tukey
HSD or Games Howell tests were used as
post hoc tests for intergroup comparisons.
The biochemical data were presented as
mean ± standard deviation (X±SD). The
Kruskal-Wallis test was preferred for analyz-
ing semiquantitative histopathologic grad-
ing data, and then the post-hoc Dunn’s test
was employed for the analysis. Statistical
results were presented as median (quartile
1- quartile 3). The significance level was set
at p<0.05.
RESULTS
Analysis of MDA, tGSH, SOD and CAT
levels in liver tissue
It can be seen from Fig. 1A that MDA
levels in liver tissue increased in the IRG
group (5.48±0.20) compared to healthy
rats (2.39±0.18) (p<0.001). The increase
in MDA was observed to be suppressed in the
TIRG (2.67±0.24) and CIRG (4.06±0.15)
(p<0.001). Compared with CE, TPP sup-
pressed the increase in MDA more effectively
(p<0.001). The MDA levels in TIRG and HG
were not significantly different (p=0.100).
In IRG, a decrease in tGSH levels
and SOD and CAT activities (3.42±0.24,
5.33±0.15, 3.78±0.12, respectively) were
detected with the increase in MDA compared
to HG (6.34±0.18, 9.19±0.12, 7.51±0.21,
respectively) (p<0.001). TPP (5.80±0.10,
8.76±0.13, 7.09±0.26, respectively) and CE
(4.48±0.09, 6.66±0.16, 5.39±0.37, respec-
tively) inhibited the decrease in tGSH levels
and SOD and CAT activities (p<0.001). TPP
treatment prevented the decrease in antioxi-
dants more effectively than CE (p<0.001)
(Fig. 1B-D).
Analysis of NF-κB, TNF-α, IL-1β and IL-6
levels in liver tissue
As presented in Fig. 2A-D, the levels of
NF-κB, TNF-α, IL-1β, and IL-6 in the liver tis-
sues of rats in the IRG group (4.86±0.09,
4.68±0.09, 6.29±0.15, 5.70±0.12, respec-
tively) were higher than those in the HG
group (2.38±0.11, 2.13±0.07, 3.37±0.19,
2.49±0.24, respectively) (p<0.001). Com-
pared to the IRG group, both TPP (2.57±0.12,
2.37±0.11, 3.68±0.26, 2.77±0.11, respec-
tively) and CE (3.25±0.15, 3.31±0.14,
5.00±0.48, 4.02±0.22, respectively) inhib-
ited these increases in NF-κB, TNF-α, IL-1β
and IL-6 (p<0.05). However, this inhibition
was more significant in the TPP than in the
CE group (p<0.05). There were similari-
ties between TIRG and HG in terms of NF-
κB (p=0.315), IL-1β (p=0.157) and IL-6
(p=0.064) levels.
Analysis of ALT and AST activities
in serum samples
As presented in Fig. 3A-B, ALT and AST
activities measured from the serum of rats in
the IRG (86.67±4.55, 220.00±7.72, respec-
tively) were found to be increased according
to HG (29.17±4.02, 39.17±4.92, respec-
326 Yeter et al.Investigación Clínica 65(3): 2024
tively) (p<0.001). The increase in ALT and
AST activities in the IRG appeared to be sup-
pressed in TIRG (32.83±3.55, 45.83±4.62,
respectively) and CIRG (56.50±2.43,
92.50±4.51, respectively) (p<0.001). TPP
inhibited this increase in ALT and AST ac-
tivities more effectively than CE (p<0.001).
Serum ALT (p=0.165) and AST (p=0.200)
activities of TIRG and HG were similar.
Histopathologic findings
There were normal central arteries in
the liver tissue sections, radially arranged
hepatic cords in the lobules, and normal tis-
sue structure in the HG group, as presented
in Fig. 4A and Table 1.
Upon examination of the liver tissue
of the IRG group, it was observed that the
hepatocytes around the central artery had
lost their typical morphology, were degen-
erating, their nuclei bulged and appeared
pyknotic on occasion. A noticeable phenom-
enon was the loss of radial arrangement of
hepatic cords, which resulted in irregular
and spiralized cords. There was a marked
and moderate degree of congestion in both
the central artery and surrounding blood
vessels. A significant increase was observed
in the number of Kupffer cells. Many PMNLs
were detected infiltrating the pericapillary
area (Fig. 4B and Table 1).
Notably, in the TIRG group treated with
TPP, the arrangement of hepatic cords and
hepatocytes was generally normal, and the
Kupffer cell population was relatively simi-
lar to the control group. PMNL infiltration
was rare throughout the tissue (Fig. 4C and
Table 1).
Fig. 1(A-D). Results of analyses of MDA (A), tGSH (B), SOD (C) and CAT (D) data measured from liver
tissues. *; p<0.001 vs IRG, # p>0.05 vs HG. MDA: malondialdehyde, tGSH: total glutathione,
SOD: superoxide dismutase, CAT: catalase, HG: healthy group, IRG: isoniazid+rifampicin applied
group, TIRG: thiamine pyrophosphate+izoniazid+rifampisin applied group, CIRG: Cinnamon
extract+izoniazid+rifampisin applied group.
tively) (p<0.001). The increase in ALT and
AST activities in the IRG appeared to be sup-
pressed in TIRG (32.83±3.55, 45.83±4.62,
respectively) and CIRG (56.50±2.43,
92.50±4.51, respectively) (p<0.001). TPP
inhibited this increase in ALT and AST ac-
tivities more effectively than CE (p<0.001).
Serum ALT (p=0.165) and AST (p=0.200)
activities of TIRG and HG were similar.
Histopathologic findings
There were normal central arteries in
the liver tissue sections, radially arranged
hepatic cords in the lobules, and normal tis-
sue structure in the HG group, as presented
in Fig. 4A and Table 1.
Upon examination of the liver tissue
of the IRG group, it was observed that the
hepatocytes around the central artery had
lost their typical morphology, were degen-
erating, their nuclei bulged and appeared
pyknotic on occasion. A noticeable phenom-
enon was the loss of radial arrangement of
hepatic cords, which resulted in irregular
and spiralized cords. There was a marked
and moderate degree of congestion in both
the central artery and surrounding blood
vessels. A significant increase was observed
in the number of Kupffer cells. Many PMNLs
were detected infiltrating the pericapillary
area (Fig. 4B and Table 1).
Notably, in the TIRG group treated with
TPP, the arrangement of hepatic cords and
hepatocytes was generally normal, and the
Kupffer cell population was relatively simi-
lar to the control group. PMNL infiltration
was rare throughout the tissue (Fig. 4C and
Table 1).
Fig. 1(A-D). Results of analyses of MDA (A), tGSH (B), SOD (C) and CAT (D) data measured from liver
tissues. *; p<0.001 vs IRG, # p>0.05 vs HG. MDA: malondialdehyde, tGSH: total glutathione,
SOD: superoxide dismutase, CAT: catalase, HG: healthy group, IRG: isoniazid+rifampicin applied
group, TIRG: thiamine pyrophosphate+izoniazid+rifampisin applied group, CIRG: Cinnamon
extract+izoniazid+rifampisin applied group.
Thiamine pyrophosphate and cinnamon protect rats from induced liver damage 327Vol. 65(3): 321 - 334, 2024
Fig. 2 (A-D). Results of analyses of NF-κB (A), IL-1β (B), TNF-α (C), and IL-6 (D) data measured from liver
tissues. *; p<0.05 vs IRG, #, p>0.05 vs HG.
NF-κB: nuclear factor kappa-B, IL-1β: interleukin 1-beta, TNF-α: tumor necrosis factor-alpha, IL-6:
interleukin-6, HG: healthy group, IRG: isoniazid+rifampicin group, TIRG: thiamine pyrophosphate
+izoniazid+rifampisin group, CIRG: Cinnamon extract+izoniazid+rifampisin group.
Fig. 3 (A-B). Results of analyses of ALT (A) and AST (B) data measured from serum. *; p<0.001 vs IRG, #,
p>0.05 vs HG.
ALT: alanine aminotransferase, AST: aspartate aminotransferase HG: healthy group, IRG:
isoniazid+rifampicin group, TIRG: thiamine pyrophosphate+izoniazid+rifampisin group, CIRG:
cinnamon extract+izoniazid+rifampisin group.
Fig. 2 (A-D). Results of analyses of NF-κB (A), IL-1β (B), TNF-α (C), and IL-6 (D) data measured from liver
tissues. *; p<0.05 vs IRG, #, p>0.05 vs HG.
NF-κB: nuclear factor kappa-B, IL-1β: interleukin 1-beta, TNF-α: tumor necrosis factor-alpha, IL-6:
interleukin-6, HG: healthy group, IRG: isoniazid+rifampicin group, TIRG: thiamine pyrophosphate
+izoniazid+rifampisin group, CIRG: Cinnamon extract+izoniazid+rifampisin group.
Fig. 3 (A-B). Results of analyses of ALT (A) and AST (B) data measured from serum. *; p<0.001 vs IRG, #,
p>0.05 vs HG.
ALT: alanine aminotransferase, AST: aspartate aminotransferase HG: healthy group, IRG:
isoniazid+rifampicin group, TIRG: thiamine pyrophosphate+izoniazid+rifampisin group, CIRG:
cinnamon extract+izoniazid+rifampisin group.
328 Yeter et al.Investigación Clínica 65(3): 2024
Fig. 4 (A-D). A. Liver tissue belonging to the HG. ; hepatocyte, ; Kupffer cell, ; normal appearance of
blood vessels (H&E x200). B. Liver tissue belonging to the IRG. ; degenerated hepatocyte, ; irreg-
ularity in hepatic cords, ; increased Kupffer cells, ; polymorphonuclear cell infiltration, ; moder-
ately congested blood vessel (H&E x 200). C. Liver tissue belonging to the TIRG. ; hepatocyte with
normal morphology, : Kupffer cell, ; slightly congested blood vessel (H&E x 200). D. Liver tissue
belonging to the CIRG. ; moderately degenerated hepatocytes, ; increased Kupffer cells, ; irreg-
ularity in the hepatic cords, ; weakly polymorphonuclear cell infiltration, ; moderately congested
blood vessels (H&E x200). HG: healthy group, IRG: isoniazid+rifampicin group, TIRG: thiamine
pyrophosphate+izoniazid+rifampisin group, CIRG: Cinnamon extract+izoniazid+rifampisin group.
Table 1
Analysis results of histopathological grading data of liver tissues from rats.
Fig. 4 (A-D). A. Liver tissue belonging to the HG. ; hepatocyte, ; Kupffer cell, ; normal appearance of
blood vessels (H&E x200). B. Liver tissue belonging to the IRG. ; degenerated hepatocyte, ; irreg-
ularity in hepatic cords, ; increased Kupffer cells, ; polymorphonuclear cell infiltration, ; moder-
ately congested blood vessel (H&E x 200). C. Liver tissue belonging to the TIRG. ; hepatocyte with
normal morphology, : Kupffer cell, ; slightly congested blood vessel (H&E x 200). D. Liver tissue
belonging to the CIRG. ; moderately degenerated hepatocytes, ; increased Kupffer cells, ; irreg-
ularity in the hepatic cords, ; weakly polymorphonuclear cell infiltration, ; moderately congested
blood vessels (H&E x200). HG: healthy group, IRG: isoniazid+rifampicin group, TIRG: thiamine
pyrophosphate+izoniazid+rifampisin group, CIRG: Cinnamon extract+izoniazid+rifampisin group.
Table 1
Analysis results of histopathological grading data of liver tissues from rats.
Thiamine pyrophosphate and cinnamon protect rats from induced liver damage 329Vol. 65(3): 321 - 334, 2024
In the CE-treated CIRG group, it was re-
markable that hepatic cords were irregular,
and hepatocyte degeneration was at a mod-
erate level. In general, it was observed that
the congestion in the blood vessels was at a
moderate level, and the Kupffer cell popula-
tion was increased. Mild PMNL infiltration
was also determined in the pericapillary area
(Fig. 4D and Table 1).
DİSCUSSİON
The primary treatment protocol for
tuberculosis consists of isoniazid, rifampi-
cin, pyrazinamide, and ethambutol for six
months 18
. Increased dosages of these drugs
have been tried to overcome resistance,
which has increased the incidence of side
effects 19
. Drug-related side effects often re-
sult in a change in treatment, which can ad-
versely affect the treatment’s effectiveness.
One of the most common side effects of an-
tituberculosis treatment is hepatotoxicity 18
.
In the literature, mitochondrial dysfunction
and oxidative stress have been reported to be
involved in the mechanism of IRC-induced
hepatotoxicity 19
. In this study, TPP 10 and
CE 14 , both with antioxidative activity, were
examined biochemically and histopathologi-
cally for their protective effects on IRC-in-
duced hepatotoxicity. Our biochemical anal-
ysis revealed that IRC use caused an increase
in oxidant (MDA) and inflammatory markers
(NF-κB, TNF-α, IL-1β, and IL-6), a decrease
in antioxidants (tGSH, SOD, and CAT), and
an increase in ALT and AST activities.
As mentioned above, the role of LPO in
the pathogenesis of isoniazid-induced oxida-
tive liver injury was demonstrated 6
. Rifam-
picin, on the other hand, was reported to
exhibit additive and even synergistic hepa-
totoxicity when used in combination with
isoniazid 4
. The cause of this condition has
been attributed to rifampicin stimulating
hydrolases and the formation of hepatotoxic
reactive metabolites from isoniazid 8
. As is
well known, MDA is an LPO product used as
a biomarker of oxidative stress 20
. MDA ex-
hibits a toxic effect; additionally, convert-
ing ROS into active substances causes cell
membrane damage, leading to apoptosis
and liver necrosis 21
. An experimental study
has found that four weeks of IRC treatment
has increased MDA in rat livers 22
. By sup-
pressing the increase in MDA in the group
administered TPP in conjunction with IRC,
it is evident that TPP displays antioxidant
properties. It was suggested in the litera-
ture that thiamine interacted with free radi-
cals and hydroperoxides and inhibited LPO
23
. Previously, TPP was also tested in oxida-
tive liver injury induced by propofol 11 and
metazimol 24 and was found to inhibit the in-
crease in MDA levels in rat livers, similar to
our study. In our study, CE suppressed the
increase in MDA levels. However, CE sup-
pressed the increase in MDA levels less than
TPP. Moselhy et al. reported that aqueous
and ethanol extracts of cinnamon could pre-
vent the increase in MDA levels by reducing
LPO in carbon tetrachloride-induced oxida-
tive liver injury in rats 25 .
The liver tissues of the animals were
also analyzed in terms of antioxidants. For
this purpose, tissue tGSH levels and SOD and
CAT activities were determined. GSH repre-
sented the most important endogenous non-
enzymatic antioxidant. GSH plays a role in
the protection of cells from ROS.10 The lit-
erature reported that hydrazine reacted with
the sulfhydryl group of GSH, depleting its
levels in hepatocytes and causing cell death
26
. In our study, tGSH levels were found to be
decreased in the IRC group. Similarly, Zhang
et al. found that IRC decreased GSH levels in
liver tissue and serum samples 27
. During the
metabolism of antituberculosis drugs, toxic
metabolites and free radicals deplete endog-
enous antioxidants, making the liver suscep-
tible to further damage 28
. According to the
literature, external administration of agents
with antioxidant activity could contribute
to tissue defense in such cases 29
. Our study
determined that TPP administered with IRC
prevented the decrease in tGSH levels and
even maintained them at healthy levels for
In the CE-treated CIRG group, it was re-
markable that hepatic cords were irregular,
and hepatocyte degeneration was at a mod-
erate level. In general, it was observed that
the congestion in the blood vessels was at a
moderate level, and the Kupffer cell popula-
tion was increased. Mild PMNL infiltration
was also determined in the pericapillary area
(Fig. 4D and Table 1).
DİSCUSSİON
The primary treatment protocol for
tuberculosis consists of isoniazid, rifampi-
cin, pyrazinamide, and ethambutol for six
months 18
. Increased dosages of these drugs
have been tried to overcome resistance,
which has increased the incidence of side
effects 19
. Drug-related side effects often re-
sult in a change in treatment, which can ad-
versely affect the treatment’s effectiveness.
One of the most common side effects of an-
tituberculosis treatment is hepatotoxicity 18
.
In the literature, mitochondrial dysfunction
and oxidative stress have been reported to be
involved in the mechanism of IRC-induced
hepatotoxicity 19
. In this study, TPP 10 and
CE 14 , both with antioxidative activity, were
examined biochemically and histopathologi-
cally for their protective effects on IRC-in-
duced hepatotoxicity. Our biochemical anal-
ysis revealed that IRC use caused an increase
in oxidant (MDA) and inflammatory markers
(NF-κB, TNF-α, IL-1β, and IL-6), a decrease
in antioxidants (tGSH, SOD, and CAT), and
an increase in ALT and AST activities.
As mentioned above, the role of LPO in
the pathogenesis of isoniazid-induced oxida-
tive liver injury was demonstrated 6
. Rifam-
picin, on the other hand, was reported to
exhibit additive and even synergistic hepa-
totoxicity when used in combination with
isoniazid 4
. The cause of this condition has
been attributed to rifampicin stimulating
hydrolases and the formation of hepatotoxic
reactive metabolites from isoniazid 8
. As is
well known, MDA is an LPO product used as
a biomarker of oxidative stress 20
. MDA ex-
hibits a toxic effect; additionally, convert-
ing ROS into active substances causes cell
membrane damage, leading to apoptosis
and liver necrosis 21
. An experimental study
has found that four weeks of IRC treatment
has increased MDA in rat livers 22
. By sup-
pressing the increase in MDA in the group
administered TPP in conjunction with IRC,
it is evident that TPP displays antioxidant
properties. It was suggested in the litera-
ture that thiamine interacted with free radi-
cals and hydroperoxides and inhibited LPO
23
. Previously, TPP was also tested in oxida-
tive liver injury induced by propofol 11 and
metazimol 24 and was found to inhibit the in-
crease in MDA levels in rat livers, similar to
our study. In our study, CE suppressed the
increase in MDA levels. However, CE sup-
pressed the increase in MDA levels less than
TPP. Moselhy et al. reported that aqueous
and ethanol extracts of cinnamon could pre-
vent the increase in MDA levels by reducing
LPO in carbon tetrachloride-induced oxida-
tive liver injury in rats 25 .
The liver tissues of the animals were
also analyzed in terms of antioxidants. For
this purpose, tissue tGSH levels and SOD and
CAT activities were determined. GSH repre-
sented the most important endogenous non-
enzymatic antioxidant. GSH plays a role in
the protection of cells from ROS.10 The lit-
erature reported that hydrazine reacted with
the sulfhydryl group of GSH, depleting its
levels in hepatocytes and causing cell death
26
. In our study, tGSH levels were found to be
decreased in the IRC group. Similarly, Zhang
et al. found that IRC decreased GSH levels in
liver tissue and serum samples 27
. During the
metabolism of antituberculosis drugs, toxic
metabolites and free radicals deplete endog-
enous antioxidants, making the liver suscep-
tible to further damage 28
. According to the
literature, external administration of agents
with antioxidant activity could contribute
to tissue defense in such cases 29
. Our study
determined that TPP administered with IRC
prevented the decrease in tGSH levels and
even maintained them at healthy levels for
330 Yeter et al.Investigación Clínica 65(3): 2024
the healthy group. Delen et al. similarly con-
cluded that TPP inhibited the depletion of
tGSH in the liver caused by propofol use 11.
The decrease in tGSH levels in CE-treated
animals was suppressed, although not as
much as in TPP-treated animals. In a previ-
ous study, CE was found to possess antioxi-
dant properties and to suppress GSH deple-
tion in liver tissue 29
.
Results of our analysis indicated that
SOD and CAT activities were also decreased
in the IRC group in which tGSH levels were
low. By accelerating the conversion of oxygen
to hydrogen peroxide (H2
O2
), SOD was the
first antioxidant enzyme to fight ROS 27
. CAT,
on the other hand, converted the toxic H2
O2
into H2
O and oxygen 30
. As a result, SOD and
CAT acted as mutually supportive antioxidant
enzymes that protected ROS 27
. It was dem-
onstrated in previous preclinical studies that
IRC decreased the activity of SOD and CAT in
the liver 31,32
. The results of this study revealed
that TPP treatment was able to prevent the
decline in SOD and CAT activities. Through-
out the body, TPP catalyzes various chemical
reactions. As a cofactor of enzymes involved
in maintaining cell redox, it synthesized re-
duced nicotinamide adenine dinucleotide
phosphate and glutathione and increased the
synthesis of antioxidants 24
. In addition, Bedir
et al. found that TPP prevented metamizole-
induced decreases in the activities of SOD
and CAT in the liver tissues of rats 24
. El-Kholy
et al. used CE, another therapeutic drug, to
treat oxidative liver damage induced by amox-
icillin/clavulanate and found that it inhibited
the decrease in SOD and CAT activity, similar
to our results 29
.
Previously, it was reported that IRC
treatment caused inflammation. This leads
to activation of macrophages and infiltra-
tion of circulating immune cells into the tis-
sue 28
. In contrast, sterile inflammation also
contributed to hepatotoxicity by increasing
oxidative stress. NF-κB was a transcription
factor that modulated inflammation 28,33
. Ac-
cording to our biochemical results and previ-
ous studies, increased NF-κB levels with IRC
treatment confirmed inflammatory activity
28,31
. In the TPP-treated group, the increase
in NF-κB levels seemed to be prevented. Sim-
ilarly, Ozer et al. found that TPP suppressed
the increase of NF-κB in damaged ovarian
tissue 34
. In our CE-treated group, NF-κB
levels were higher than the TPP group and
lower than the IRC group. A recent study
found that this increase was suppressed in
obese rats treated with Cinnamon powder
compared to rats with increased hepatic
NF-κB expression with obesity 35
. Our study
also investigated liver tissues for the levels
of proinflammatory cytokines such as TNF-α,
IL-1β, and IL-6.
Increased ROS released inactive NF-κB
through a series of reactions. Activated NF-
κB induced gene transcription of TNF- α and
IL-1β 21
. On the other hand, TNF-α, which
is considered the primary mediator induc-
ing the inflammatory response, has been re-
ported to activate NF-κB by binding to the
TNF receptor 31,36
. In addition, it was pointed
out that TNF-α increased ROS production
that triggered liver injury through various
pathways 31 . IL-1β had intense proinflamma-
tory activity like TNF-α 37
. Although IL-6 also
prevented acute liver inflammation, its pro-
longed stimulation resulted in tissue dam-
age to the liver 38
. Our study discovered that
TNF-α, IL-1β, and IL-6 levels were similarly
increased in the IRC group in which NF-κB
levels were increased. Patel et al. also found
an increase in TNF-α, IL-1β, and IL-6 levels
in their study on the effects of IRC on the
liver 38
. Cytokine levels in rats administered
with TPP and CE were lower than those of the
IRC group. No studies have been conducted
in the literature on the effect of TPP on tis-
sue cytokine levels in oxidative liver injury.
However, it was reported that TPP prevented
the increase in TNF-α and IL-1β levels due
to ethanol toxicity in optic nerve tissue 39
.
In the literature, studies also reported that
CE, another therapeutic agent, exhibited
anti-inflammatory activity by suppressing
the increase in TNF-α, IL-1β, and IL-6 levels
induced by various causes in the liver 40,41
.
the healthy group. Delen et al. similarly con-
cluded that TPP inhibited the depletion of
tGSH in the liver caused by propofol use 11.
The decrease in tGSH levels in CE-treated
animals was suppressed, although not as
much as in TPP-treated animals. In a previ-
ous study, CE was found to possess antioxi-
dant properties and to suppress GSH deple-
tion in liver tissue 29
.
Results of our analysis indicated that
SOD and CAT activities were also decreased
in the IRC group in which tGSH levels were
low. By accelerating the conversion of oxygen
to hydrogen peroxide (H2
O2
), SOD was the
first antioxidant enzyme to fight ROS 27
. CAT,
on the other hand, converted the toxic H2
O2
into H2
O and oxygen 30
. As a result, SOD and
CAT acted as mutually supportive antioxidant
enzymes that protected ROS 27
. It was dem-
onstrated in previous preclinical studies that
IRC decreased the activity of SOD and CAT in
the liver 31,32
. The results of this study revealed
that TPP treatment was able to prevent the
decline in SOD and CAT activities. Through-
out the body, TPP catalyzes various chemical
reactions. As a cofactor of enzymes involved
in maintaining cell redox, it synthesized re-
duced nicotinamide adenine dinucleotide
phosphate and glutathione and increased the
synthesis of antioxidants 24
. In addition, Bedir
et al. found that TPP prevented metamizole-
induced decreases in the activities of SOD
and CAT in the liver tissues of rats 24
. El-Kholy
et al. used CE, another therapeutic drug, to
treat oxidative liver damage induced by amox-
icillin/clavulanate and found that it inhibited
the decrease in SOD and CAT activity, similar
to our results 29
.
Previously, it was reported that IRC
treatment caused inflammation. This leads
to activation of macrophages and infiltra-
tion of circulating immune cells into the tis-
sue 28
. In contrast, sterile inflammation also
contributed to hepatotoxicity by increasing
oxidative stress. NF-κB was a transcription
factor that modulated inflammation 28,33
. Ac-
cording to our biochemical results and previ-
ous studies, increased NF-κB levels with IRC
treatment confirmed inflammatory activity
28,31
. In the TPP-treated group, the increase
in NF-κB levels seemed to be prevented. Sim-
ilarly, Ozer et al. found that TPP suppressed
the increase of NF-κB in damaged ovarian
tissue 34
. In our CE-treated group, NF-κB
levels were higher than the TPP group and
lower than the IRC group. A recent study
found that this increase was suppressed in
obese rats treated with Cinnamon powder
compared to rats with increased hepatic
NF-κB expression with obesity 35
. Our study
also investigated liver tissues for the levels
of proinflammatory cytokines such as TNF-α,
IL-1β, and IL-6.
Increased ROS released inactive NF-κB
through a series of reactions. Activated NF-
κB induced gene transcription of TNF- α and
IL-1β 21
. On the other hand, TNF-α, which
is considered the primary mediator induc-
ing the inflammatory response, has been re-
ported to activate NF-κB by binding to the
TNF receptor 31,36
. In addition, it was pointed
out that TNF-α increased ROS production
that triggered liver injury through various
pathways 31 . IL-1β had intense proinflamma-
tory activity like TNF-α 37
. Although IL-6 also
prevented acute liver inflammation, its pro-
longed stimulation resulted in tissue dam-
age to the liver 38
. Our study discovered that
TNF-α, IL-1β, and IL-6 levels were similarly
increased in the IRC group in which NF-κB
levels were increased. Patel et al. also found
an increase in TNF-α, IL-1β, and IL-6 levels
in their study on the effects of IRC on the
liver 38
. Cytokine levels in rats administered
with TPP and CE were lower than those of the
IRC group. No studies have been conducted
in the literature on the effect of TPP on tis-
sue cytokine levels in oxidative liver injury.
However, it was reported that TPP prevented
the increase in TNF-α and IL-1β levels due
to ethanol toxicity in optic nerve tissue 39
.
In the literature, studies also reported that
CE, another therapeutic agent, exhibited
anti-inflammatory activity by suppressing
the increase in TNF-α, IL-1β, and IL-6 levels
induced by various causes in the liver 40,41
.
Thiamine pyrophosphate and cinnamon protect rats from induced liver damage 331Vol. 65(3): 321 - 334, 2024
In this study, the ALT and AST activi-
ties of blood samples obtained from animals
were measured. These transaminases were
sensitive markers of liver cell damage, and
these biomarkers have been widely used in
the past and present to assess liver func-
tion 42 . In both clinical and experimental
studies, it has been demonstrated that IRC
treatment increases the levels of ALT and
AST 18,26,42 . Our study also demonstrated
increased ALT and AST activity after IRC
treatment. A lower enzyme activity was ob-
served in both TPP and CE-treated groups,
but a lower activity was observed in the
TPP-treated group. As previously reported,
TPP inhibited the increase in ALT, AST, and
lactate dehydrogenase activities following
acetaminophen 43 and cisplatin-induced liv-
er injury 12 . Mosely et al. also demonstrated
that CE inhibited the increase in ALT and
AST activities 25 .
A histologic analysis of liver tissues con-
firmed our biochemical findings. Degenera-
tion of hepatocytes and marked congestion
of blood vessels were observed in the IRC
group. In addition, an increase in the num-
ber of Kupffer cells and PMNL was observed.
According to previous studies, IRC induced
changes in the liver, such as sinusoidal dila-
tation, infiltration of immune cells, inflam-
mation in the portal system, and necrosis of
the liver 27,28
. Our histopathological analysis
showed that the hepatocytes and Kupffer
cell populations of the TPP-treated group
were similar to those of the control group,
and rare evidence of PMNL infiltration was
observed. Several studies demonstrated that
TPP protected liver tissue from oxidative
damage and prevented structural changes
such as cellular degeneration, necrosis, and
infiltration of inflammatory cells 11,43
. In the
CE group, there was moderate hepatocyte
degeneration and vascular congestion. In
addition, there was a moderate increase in
the Kupffer cell population and mild PMNL
infiltration. However, some previous studies
reported that CE treatment altogether 44 or
almost entirely 25 prevented tissue damage
in oxidative liver injury. However, our his-
topathologic evaluation indicated that TPP
protected the histologic structure of the liv-
er better than CE, similar to our biochemi-
cal results.
Limitations
In the future, total oxidant levels and
antioxidant levels should be measured to in-
vestigate the mechanism of hepatoprotec-
tive effect of TPP and CE in greater detail.
Additionally, we believe that anti-inflamma-
tory cytokine levels should be evaluated.
Our results revealed that IRC induced
an increase in oxidants and proinflammato-
ry cytokines and a decrease in antioxidants
in the liver tissues of the animals. More-
over, IRC treatment caused an increase in
liver function parameters and damage to
the histological structure of the liver. TPP
and CE inhibited both biochemical and his-
topathologic changes. However, TPP pro-
tected liver tissue from IRC-induced dam-
age better than CE. The results of our study
indicated that adding TPP to the treatment
might be an effective therapeutic strategy
for preventing IRC-induced hepatotoxicity.
Moreover, based on the results of our litera-
ture review, our study was the first to com-
pare the antioxidant and anti-inflammatory
properties of TPP and CE. As a result of our
current study, we are likely to contribute to
future experimental and clinical studies on
diseases where oxidative stress and inflam-
mation are involved in the pathogenesis.
Competing interest
The authors declared no conflict of in-
terest.
Data availability
Study data can be obtained from the
corresponding author upon request.
Funding
None.
In this study, the ALT and AST activi-
ties of blood samples obtained from animals
were measured. These transaminases were
sensitive markers of liver cell damage, and
these biomarkers have been widely used in
the past and present to assess liver func-
tion 42 . In both clinical and experimental
studies, it has been demonstrated that IRC
treatment increases the levels of ALT and
AST 18,26,42 . Our study also demonstrated
increased ALT and AST activity after IRC
treatment. A lower enzyme activity was ob-
served in both TPP and CE-treated groups,
but a lower activity was observed in the
TPP-treated group. As previously reported,
TPP inhibited the increase in ALT, AST, and
lactate dehydrogenase activities following
acetaminophen 43 and cisplatin-induced liv-
er injury 12 . Mosely et al. also demonstrated
that CE inhibited the increase in ALT and
AST activities 25 .
A histologic analysis of liver tissues con-
firmed our biochemical findings. Degenera-
tion of hepatocytes and marked congestion
of blood vessels were observed in the IRC
group. In addition, an increase in the num-
ber of Kupffer cells and PMNL was observed.
According to previous studies, IRC induced
changes in the liver, such as sinusoidal dila-
tation, infiltration of immune cells, inflam-
mation in the portal system, and necrosis of
the liver 27,28
. Our histopathological analysis
showed that the hepatocytes and Kupffer
cell populations of the TPP-treated group
were similar to those of the control group,
and rare evidence of PMNL infiltration was
observed. Several studies demonstrated that
TPP protected liver tissue from oxidative
damage and prevented structural changes
such as cellular degeneration, necrosis, and
infiltration of inflammatory cells 11,43
. In the
CE group, there was moderate hepatocyte
degeneration and vascular congestion. In
addition, there was a moderate increase in
the Kupffer cell population and mild PMNL
infiltration. However, some previous studies
reported that CE treatment altogether 44 or
almost entirely 25 prevented tissue damage
in oxidative liver injury. However, our his-
topathologic evaluation indicated that TPP
protected the histologic structure of the liv-
er better than CE, similar to our biochemi-
cal results.
Limitations
In the future, total oxidant levels and
antioxidant levels should be measured to in-
vestigate the mechanism of hepatoprotec-
tive effect of TPP and CE in greater detail.
Additionally, we believe that anti-inflamma-
tory cytokine levels should be evaluated.
Our results revealed that IRC induced
an increase in oxidants and proinflammato-
ry cytokines and a decrease in antioxidants
in the liver tissues of the animals. More-
over, IRC treatment caused an increase in
liver function parameters and damage to
the histological structure of the liver. TPP
and CE inhibited both biochemical and his-
topathologic changes. However, TPP pro-
tected liver tissue from IRC-induced dam-
age better than CE. The results of our study
indicated that adding TPP to the treatment
might be an effective therapeutic strategy
for preventing IRC-induced hepatotoxicity.
Moreover, based on the results of our litera-
ture review, our study was the first to com-
pare the antioxidant and anti-inflammatory
properties of TPP and CE. As a result of our
current study, we are likely to contribute to
future experimental and clinical studies on
diseases where oxidative stress and inflam-
mation are involved in the pathogenesis.
Competing interest
The authors declared no conflict of in-
terest.
Data availability
Study data can be obtained from the
corresponding author upon request.
Funding
None.
332 Yeter et al.Investigación Clínica 65(3): 2024
ORCID numbers
• Bahtinur Yeter (BY):
0000-0003-0336-8161
• Renad Mammadov (RM):
0000-0002-5785-1960
• Zeynep Koc (ZK):
0000-0002-0716-7773
• Seval Bulut (SB):
0000-0003-4992-1241
• Tugba Bal Tastan(TBT):
0000-0001-8257-8639
• Mine Gulaboglu (MG):
0000-0002-3248-1502
• Halis Suleyman (HS):
0000-0002-9239-4099
Author Contributions
BY: research concept and design, writ-
ing the article, critical revision of the arti-
cle, final approval; RM: collection and/or as-
sembly of data, final approval; ZK: collection
and/or assembly of data, final approval; SB:
data analysis and interpretation, writing the
article, critical revision of the article, final
approval; TBT: collection and/or assembly
of data, final approval; MG: collection and/
or assembly of data, final approval; HS: re-
search concept and design, writing the ar-
ticle, final approval.
REFERENCES
1. Combrink M, du Preez I. Metabolomics
describes previously unknown toxicity me-
chanisms of isoniazid and rifampicin. Toxi-
col Lett 2020;322:104-110.
2. Suárez I, Fünger SM, Kröger S, Radema-
cher J, Fätkenheuer G, Rybniker J. The
diagnosis and treatment of tuberculosis.
Dtsch Arztebl Int 2019;116:729-735.
3. Forget EJ, Menzies D. Adverse reactions
to first-line antituberculosis drugs. Expert
Opin Drug Saf 2006;5:231-249.
4. Yew WW, Chang KC, Chan DP. Oxidative
stress and first-line antituberculosis drug-
induced hepatotoxicity. Antimicrob Agents
Chemother 2018;62:e02637-17
5. Song L, Zhang ZR, Zhang JL, Zhu XB, He
L, Shi Z, Gao L, Li Y, Hu B, Feng FM. Mi-
croRNA-122 is involved in oxidative stress
in isoniazid-induced liver injury in mice.
Genet Mol Res 2015;14:13258-13265.
6. Ahadpour M, Eskandari MR, Mashayekhi
V, Haj Mohammad Ebrahim Tehrani K, Ja-
farian I, Naserzadeh P, Hosseini MJ. Mito-
chondrial oxidative stress and dysfunction
induced by isoniazid: study on isolated rat
liver and brain mitochondria. Drug Chem
Toxicol 2016;39:224-232.
7. Shuhendler AJ, Pu K, Cui L, Uetrecht JP,
Rao J. Real-time imaging of oxidative and
nitrosative stress in the liver of live ani-
mals for drug-toxicity testing. Nat Biote-
chnol 2014;32:373-380.
8. Askgaard DS, Wilcke T, Døssing M. He-
patotoxicity caused by the combined ac-
tion of isoniazid and rifampicin. Thorax
1995;50:213-214.
9. Sica DA. Loop diuretic therapy, thiamine
balance, and heart failure. Congest Heart
Fail 2007;13:244-247.
10. Altuner D, Cetin N, Suleyman B, Aslan Z,
Hacimuftuoglu A, Gulaboglu M, Isaoglu
N, I Demiryilmaz I, Suleyman H. Effect of
thiamine pyrophosphate on ischemia-reper-
fusion induced oxidative damage in rat kid-
ney. Indian J Pharmacol 2013;45:339-343.
11. Acun Delen L, Korkmaz Dişli Z, Taş HG,
Kuyrukluyildiz U, Yazici GN, Süleyman
B, Kuzucu M, Gürsu C, Suleyman H.
The effects of thiamine pyrophosphate on
propofol-induced oxidative liver injury and
effect on dysfunction. Gen Physiol Biophys
2022;41:63-70.
12. Turan MI, Cayir A, Cetin N, Suleyman H,
Siltelioglu Turan I, Tan H. An investiga-
tion of the effect of thiamine pyrophospha-
te on cisplatin-induced oxidative stress and
DNA damage in rat brain tissue compared
with thiamine: thiamine and thiamine
pyrophosphate effects on cisplatin neuro-
toxicity. Hum Exp Toxicol 2014;33:14-21.
13. Yanakiev S. Effects of cinnamon (Cinna-
momum spp.) in dentistry: A review. Mole-
cules 2020;25:4184.
ORCID numbers
• Bahtinur Yeter (BY):
0000-0003-0336-8161
• Renad Mammadov (RM):
0000-0002-5785-1960
• Zeynep Koc (ZK):
0000-0002-0716-7773
• Seval Bulut (SB):
0000-0003-4992-1241
• Tugba Bal Tastan(TBT):
0000-0001-8257-8639
• Mine Gulaboglu (MG):
0000-0002-3248-1502
• Halis Suleyman (HS):
0000-0002-9239-4099
Author Contributions
BY: research concept and design, writ-
ing the article, critical revision of the arti-
cle, final approval; RM: collection and/or as-
sembly of data, final approval; ZK: collection
and/or assembly of data, final approval; SB:
data analysis and interpretation, writing the
article, critical revision of the article, final
approval; TBT: collection and/or assembly
of data, final approval; MG: collection and/
or assembly of data, final approval; HS: re-
search concept and design, writing the ar-
ticle, final approval.
REFERENCES
1. Combrink M, du Preez I. Metabolomics
describes previously unknown toxicity me-
chanisms of isoniazid and rifampicin. Toxi-
col Lett 2020;322:104-110.
2. Suárez I, Fünger SM, Kröger S, Radema-
cher J, Fätkenheuer G, Rybniker J. The
diagnosis and treatment of tuberculosis.
Dtsch Arztebl Int 2019;116:729-735.
3. Forget EJ, Menzies D. Adverse reactions
to first-line antituberculosis drugs. Expert
Opin Drug Saf 2006;5:231-249.
4. Yew WW, Chang KC, Chan DP. Oxidative
stress and first-line antituberculosis drug-
induced hepatotoxicity. Antimicrob Agents
Chemother 2018;62:e02637-17
5. Song L, Zhang ZR, Zhang JL, Zhu XB, He
L, Shi Z, Gao L, Li Y, Hu B, Feng FM. Mi-
croRNA-122 is involved in oxidative stress
in isoniazid-induced liver injury in mice.
Genet Mol Res 2015;14:13258-13265.
6. Ahadpour M, Eskandari MR, Mashayekhi
V, Haj Mohammad Ebrahim Tehrani K, Ja-
farian I, Naserzadeh P, Hosseini MJ. Mito-
chondrial oxidative stress and dysfunction
induced by isoniazid: study on isolated rat
liver and brain mitochondria. Drug Chem
Toxicol 2016;39:224-232.
7. Shuhendler AJ, Pu K, Cui L, Uetrecht JP,
Rao J. Real-time imaging of oxidative and
nitrosative stress in the liver of live ani-
mals for drug-toxicity testing. Nat Biote-
chnol 2014;32:373-380.
8. Askgaard DS, Wilcke T, Døssing M. He-
patotoxicity caused by the combined ac-
tion of isoniazid and rifampicin. Thorax
1995;50:213-214.
9. Sica DA. Loop diuretic therapy, thiamine
balance, and heart failure. Congest Heart
Fail 2007;13:244-247.
10. Altuner D, Cetin N, Suleyman B, Aslan Z,
Hacimuftuoglu A, Gulaboglu M, Isaoglu
N, I Demiryilmaz I, Suleyman H. Effect of
thiamine pyrophosphate on ischemia-reper-
fusion induced oxidative damage in rat kid-
ney. Indian J Pharmacol 2013;45:339-343.
11. Acun Delen L, Korkmaz Dişli Z, Taş HG,
Kuyrukluyildiz U, Yazici GN, Süleyman
B, Kuzucu M, Gürsu C, Suleyman H.
The effects of thiamine pyrophosphate on
propofol-induced oxidative liver injury and
effect on dysfunction. Gen Physiol Biophys
2022;41:63-70.
12. Turan MI, Cayir A, Cetin N, Suleyman H,
Siltelioglu Turan I, Tan H. An investiga-
tion of the effect of thiamine pyrophospha-
te on cisplatin-induced oxidative stress and
DNA damage in rat brain tissue compared
with thiamine: thiamine and thiamine
pyrophosphate effects on cisplatin neuro-
toxicity. Hum Exp Toxicol 2014;33:14-21.
13. Yanakiev S. Effects of cinnamon (Cinna-
momum spp.) in dentistry: A review. Mole-
cules 2020;25:4184.
Thiamine pyrophosphate and cinnamon protect rats from induced liver damage 333Vol. 65(3): 321 - 334, 2024
14. Gruenwald J, Freder J, Armbruester N.
Cinnamon and health. Crit Rev Food Sci
Nutr 2010;50:822-834.
15. Dorri M, Hashemitabar S, Hosseinzadeh
H. Cinnamon (Cinnamomum zeylanicum)
as an antidote or a protective agent against
natural or chemical toxicities: a review.
Drug Chem Toxicol 2018;41:338-351.
16. Ho SC, Chang KS, Chang PW. Inhibi-
tion of neuroinflammation by cinnamon
and its main components. Food Chem
2013;138:2275-2282.
17. Góth L. A simple method for determina-
tion of serum catalase activity and revi-
sion of reference range. Clin Chim Acta
1991;196(2-3):143-151.
18. Tostmann A, Boeree MJ, Aarnoutse RE,
de Lange WC, van der Ven AJ, Dekhuijzen
R. Antituberculosis drug-induced hepato-
toxicity: concise up-to-date review. J Gas-
troenterol Hepatol 2008;23(2):192-202.
19. Enriquez-Cortina C, Almonte-Becerril M,
Clavijo-Cornejo D, Palestino-Domínguez
M, Bello-Monroy O, Nuño N, López A,
Bucio L, Souza V, Hernández-Pando R,
Muñoz L, Gutiérrez-Ruiz MC, Gómez-
Quiroz LE. Hepatocyte growth factor
protects against isoniazid/rifampicin-in-
duced oxidative liver damage. Toxicol Sci
2013;135:26-36.
20. Aynaoglu Yildiz G, Yapca OE, Dinc K,
Gursul C, Gundogdu B, Aktas M, Suley-
man Z, Bulut S, Suleyman H. Stress-asso-
ciated ovarian damage, infertility, and de-
lay in achieving pregnancy and treatment
options. Invest Clin 2023;64:513-523. doi.
org/10.54817/IC.v64n4a08
21. Zhuang X, Li L, Liu T, Zhang R, Yang P,
Wang X, Dai L. Mechanisms of isoniazid
and rifampicin-induced liver injury and the
effects of natural medicinal ingredients: A
review. Front Pharmacol 2022;13:1037814.
22. Pelapelapon AA, Rohmawaty E, Her-
man H. Evaluation of the hepatoprotec-
tive effect of Plantago major extract in a
rifampicin-isoniazid induced hepatitis rat
model. Trends in Sciences 2023;20:6331-
6331.
23. Sarandol E, Tas S, Serdar Z, Dirican M.
Effects of thiamine treatment on oxidative
stress in experimental diabetes. Bratislavs-
ke Lekarske Listy 2020;121:235-241.
24. Bedir Z, Erdem KTO, Can A, Çiçek B,
Gülaboğlu M, Süleyman Z, Gürsul C, Mo-
khtare B, Özçiçek F, Süleyman H. Effect
of thiamine pyrophosphate upon possible
metamizole-induced liver injury in rats. Int
J Pharmacol 2023;19:139-146.
25. Moselhy SS, Ali HK. Hepatoprotective
effect of cinnamon extracts against carbon
tetrachloride induced oxidative stress and
liver injury in rats. Biol Res 2009;42:93-98.
26. Naji KM, Al-Khatib BY, Al-Haj NS, D’souza
MR. Hepatoprotective activity of melittin
on isoniazid-and rifampicin-induced liver
injuries in male albino rats. BMC Pharma-
col Toxicol 2021;22:1-11.
27. Zhang G, Zhu J, Zhou Y, Wei Y, Xi L, Qin
H, Rao Z, Han M, Ma Y, Wu X. Hesperi-
din alleviates oxidative stress and upre-
gulates the multidrug resistance protein
2 in isoniazid and rifampicin‐induced li-
ver injury in rats. J Biochem Mol Toxicol
2016;30:342-349.
28. Sanjay S, Girish C, Toi PC, Bobby Z. Gallic
acid attenuates isoniazid and rifampicin-
induced liver injury by improving hepatic
redox homeostasis through influence on
Nrf2 and NF-κB signalling cascades in Wis-
tar Rats. J Pharm Pharmacol 2021;73:473-
486.
29. El-Kholy M, Faried A, Ghada M. Role of
cinnamon extract in the protection aga-
inst amoxicillin/clavulanate-induced liver
damage in rats. IOSR-JPBS. 2019;14:14-
21.
30. Kaya A, Ceylan AF, Kavutcu M, Santa-
maria A, Šoltésová Prnová M, Stefek M,
Karasu Ç. A dual-acting aldose reductase
inhibitor impedes oxidative and carbon-
yl stress in tissues of fructose-and strep-
tozotocin-induced rats: comparison with
antioxidant stobadine. Drug ChemToxicol
2023;5:1-11.
31. He X, Song Y, Wang L, Xu J. Protective
effect of pyrrolidine dithiocarbamate on
isoniazid/rifampicin‑induced liver injury
in rats. Mol Med Rep 2020;21:463-469.
32. Yang J, Li G, Bao X, Suo Y, Xu H, Deng Y,
Feng T, Deng G. Hepatoprotective effects
14. Gruenwald J, Freder J, Armbruester N.
Cinnamon and health. Crit Rev Food Sci
Nutr 2010;50:822-834.
15. Dorri M, Hashemitabar S, Hosseinzadeh
H. Cinnamon (Cinnamomum zeylanicum)
as an antidote or a protective agent against
natural or chemical toxicities: a review.
Drug Chem Toxicol 2018;41:338-351.
16. Ho SC, Chang KS, Chang PW. Inhibi-
tion of neuroinflammation by cinnamon
and its main components. Food Chem
2013;138:2275-2282.
17. Góth L. A simple method for determina-
tion of serum catalase activity and revi-
sion of reference range. Clin Chim Acta
1991;196(2-3):143-151.
18. Tostmann A, Boeree MJ, Aarnoutse RE,
de Lange WC, van der Ven AJ, Dekhuijzen
R. Antituberculosis drug-induced hepato-
toxicity: concise up-to-date review. J Gas-
troenterol Hepatol 2008;23(2):192-202.
19. Enriquez-Cortina C, Almonte-Becerril M,
Clavijo-Cornejo D, Palestino-Domínguez
M, Bello-Monroy O, Nuño N, López A,
Bucio L, Souza V, Hernández-Pando R,
Muñoz L, Gutiérrez-Ruiz MC, Gómez-
Quiroz LE. Hepatocyte growth factor
protects against isoniazid/rifampicin-in-
duced oxidative liver damage. Toxicol Sci
2013;135:26-36.
20. Aynaoglu Yildiz G, Yapca OE, Dinc K,
Gursul C, Gundogdu B, Aktas M, Suley-
man Z, Bulut S, Suleyman H. Stress-asso-
ciated ovarian damage, infertility, and de-
lay in achieving pregnancy and treatment
options. Invest Clin 2023;64:513-523. doi.
org/10.54817/IC.v64n4a08
21. Zhuang X, Li L, Liu T, Zhang R, Yang P,
Wang X, Dai L. Mechanisms of isoniazid
and rifampicin-induced liver injury and the
effects of natural medicinal ingredients: A
review. Front Pharmacol 2022;13:1037814.
22. Pelapelapon AA, Rohmawaty E, Her-
man H. Evaluation of the hepatoprotec-
tive effect of Plantago major extract in a
rifampicin-isoniazid induced hepatitis rat
model. Trends in Sciences 2023;20:6331-
6331.
23. Sarandol E, Tas S, Serdar Z, Dirican M.
Effects of thiamine treatment on oxidative
stress in experimental diabetes. Bratislavs-
ke Lekarske Listy 2020;121:235-241.
24. Bedir Z, Erdem KTO, Can A, Çiçek B,
Gülaboğlu M, Süleyman Z, Gürsul C, Mo-
khtare B, Özçiçek F, Süleyman H. Effect
of thiamine pyrophosphate upon possible
metamizole-induced liver injury in rats. Int
J Pharmacol 2023;19:139-146.
25. Moselhy SS, Ali HK. Hepatoprotective
effect of cinnamon extracts against carbon
tetrachloride induced oxidative stress and
liver injury in rats. Biol Res 2009;42:93-98.
26. Naji KM, Al-Khatib BY, Al-Haj NS, D’souza
MR. Hepatoprotective activity of melittin
on isoniazid-and rifampicin-induced liver
injuries in male albino rats. BMC Pharma-
col Toxicol 2021;22:1-11.
27. Zhang G, Zhu J, Zhou Y, Wei Y, Xi L, Qin
H, Rao Z, Han M, Ma Y, Wu X. Hesperi-
din alleviates oxidative stress and upre-
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Biochemical and histological effects of
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of phloridzin against isoniazid-rifampicin
induced liver injury by regulating CYP450
and Nrf2/HO-1 pathway in mice. Chem
Pharm Bull 2022;70:805-811.
33. Wang N, Jiang D, Zhou C, Han X. Alpha-
solanine inhibits endothelial inflammation
via nuclear factor kappa B signaling
pathway. Adv Clin Exp Med. 2023;32:909-
920.
34. Ozer M, Ince S, Gundogdu B, Aktas M,
Uzel K, Gursul C, Suleyman H, Suley-
man Z. Effect of thiamine pyrophosphate
on cyclophosphamide-induced oxidative
ovarian damage and reproductive dys-
function in female rats. Adv Clin Exp Med
2022;31:129-137.
35. Li B, Li J, Hu S. Cinnamon could improve
hepatic steatosis caused by a high‐fat diet
via enhancing hepatic beta‐oxidation and
inhibiting hepatic lipogenesis, oxidative
damage, and inflammation in male rats. J
Food Biochem 2022;46:e14077.
36. Zhai H, Yang B, Fu Y, Zhang D, Li Y, Huang
J. Effects of somatostatin in combination
with early hemoperfusion on inflammatory,
hemorheological and oxidative parameters
during the treatment of acute pancreati-
tis. Invest Clín. 2023;64:41-52.
37. Wang Y, Che M, Xin J, Zheng Z, Li J,
Zhang S. The role of IL-1β and TNF-α in
intervertebral disc degeneration. Biomed
Pharmacother 2020;131:110660.
38. Patel S, Chaturvedi A, Dubey N, Shrivas-
tava A, Ganeshpurkar A. Ascorbic acid
ameliorates isoniazid-rifampicin-induced
hepatocellular damage in rats. iLIVER.
2022;1:72-77.
39. Ucak T, Karakurt Y, Tasli G, Cimen FK,
Icel E, Kurt N, Ahiskali I, Süleyman H.
The effects of thiamine pyrophosphate on
ethanol induced optic nerve damage. BMC
Pharmacol Toxicol 2019;20:40.
40. Kanuri G, Weber S, Volynets V, Spruss
A, Bischoff SC, Bergheim I. Cinnamon
extract protects against acute alcohol-
induced liver steatosis in mice. J Nutr
2009;139:482-487.
41. Hussain S, Ashafaq M, Alshahrani S,
Siddiqui R, Ahmed RA, Khuwaja G, Islam
F. Cinnamon oil against acetaminophen-
induced acute liver toxicity by attenuating
inflammation, oxidative stress and apopto-
sis. Toxicol Rep 2020;7:1296-1304.
42. Yilmaz I, Demiryilmaz I, Turan M, Ce-
tin N, Gul M, Süleyman H. The effects of
thiamine and thiamine pyrophosphate on
alcohol-induced hepatic damage biomar-
kers in rats. Eur Rev Med Pharmacol Sci
2015;19:664-670.
43. Uysal HB, Dağlı B, Yılmaz M, Kahyaoğlu
F, Gökçimen A, Ömürlü İK, Demirci B.
Biochemical and histological effects of
thiamine pyrophosphate against acetami-
nophen‐induced hepatotoxicity. Basic Clin
Pharmacol Toxicol 2016;118:70-76.
44. Eidi A, Mortazavi P, Bazargan M, Za-
ringhalam J. Hepatoprotective activity of
cinnamon ethanolic extract against CCI4-
induced liver injury in rats. Excli Journal.
2012;11:495.