La timosina β4 regula la función de las células endoteliales, activando la vía AKT.
Thymosin β4 regulates endothelial cell function via activating the AKT pathway.
Abstract
The vascular eendothelial cells are highly heterogeneous and associated with numerous diseases. Thymosin β4 (Tβ4) plays pleiotropic roles in endothelial cell differentiation, migration and angiogenesis. However, the underlying mechanisms played by Tβ4 in the regulation of endothelial cells have not yet been well investigated. In the present study, Tβ4 -GFP adenovirus, transfected into human umbilical vein endothelial cells (HUVECs), and cell morphology were analyzed by fluorescence microscopy. ELISA was used to determine the concentration of Tβ4 expression. Furthermore, the effects of Tβ4 overexpression on HUVECs proliferation, apoptosis and migration were investigated. Real-time quantitative PCR and western blot were conducted to examine mRNA and protein expression in HUVECs with Tβ4 overexpression. Moreover, the underlying molecular mechanism of Tβ4 in HUVECs function was tested through treatment with LY294002, a PI3K/AKT inhibitor. Overexpression of Tβ4 increased the cell ability of HUVECs, and up-regulated the expression of the proliferation markers PCNA and Cyclin D1. In addition, overexpression of Tβ4 reduced HUVECs apoptosis, both under normoxic and hypoxic conditions. Moreover, overexpression of Tβ4 increased the ability of HUVECs to migrate through the membrane and up-regulated levels of MMP-2 and MMP-9. The use of LY294002 decreased the p-AKT (Ser473) level, which was induced by Tβ4 over-expression. Importantly, LY294002 reduced Tβ4-induced HUVECs proliferation and migration. In conclusion, our results suggest that Tβ4 is a major regulator of HUVECs function by activating the AKT signaling pathway.
Downloads
References
Chen H, Liu X, Wu Y, Wu X, Wen X, Lu Y, Zhao X. Apoptosis in HUVECs induced by microRNA-616-3p via X-linked inhibitor of apoptosis protein targeting. Exp Ther Med 2021;21(6):661.
Jiang F, Xu XR, Li WM, Xia K, Wang LF, Yang XC. Monotropein alleviates H2O2‑induced inflammation, oxidative stress and apoptosis via NF-κB/AP-1 signaling. Mol Med Rep 2020;22(6):4828-4836.
Nie J. inhibits high glucose induced apoptosis in HUVEC by targeting Rab4. Biomed Pharmaother 2020;131: p. 110662.
Mi L, Zhang Y, Xu Y, Zheng X, Zhang X, Wang Z, Xue M, Jin X. HMGB1/RAGE pro-inflammatory axis promotes vascular endothelial cell apoptosis in limb ischemia/ reperfusion injury. Biomed Pharmacother. 2019; 116:109005.
Low T, Hu S-K, Goldstein AL. Complete amino acid sequence of bovine thymosin beta 4: a thymic hormone that induces terminal deoxynucleotidyl transferase activity in thymocyte populations. Proc Natl Acad Sci 1981;78(2):1162-1166.
Dubé KN, Smart N. Thymosin β4 and the vasculature: multiple roles in development, repair and protection against disease. Expert Opin Biol Ther 2018;18(sup1):131-139.
Kupatt C, Horstkotte J, Vlastos GA, Pfosser A, Lebherz C, Semisch M, Thalgott M, Büttner K, Browarzyk C, Mages J. Embryonic endothelial progenitor cells expressing a broad range of proangiogenic and remodeling factors enhance vascularization and tissue recovery in acute and chronic ischemia. The FASEB J 2005;19(11):1576- 1578.
Smart N, Risebro CA, Clark JE, Ehler E, Miquerol L, Rossdeutsch A, Marber MS, Riley PR. Thymosin β4 facilitates epicardial neovascularization of the injured adult heart. Ann N Y Acad Sci 2010;1194(1):97- 104.
Smart N, Risebro CA, Melville AA, Moses K, Schwartz RJ, Chien KR, Riley PR. Thymosin β4 induces adult epicardial progenitor mobilization and neovascularization. Nature 2007;445(7124):177-182.
Smart N, Risebro CA, Melville AA, Moses K, Schwartz RJ, Chien KR, Riley PR. Thymosin β4 is essential for coronary vessel development and promotes neovascularization via adult epicardium. Ann N Y Acad Sci 2007;1112(1):171-188.
Wyczółkowska J, Walczak-Drzewiecka A, Wagner W, Dastych J. Thymosin β4 and thymosin β4-derived peptides induce mast cell exocytosis. Peptides 2007;28(4):752-759.
Sun C, Feng SB, Cao ZW, Bei JJ, Chen Q, Xu XJ, Zhou Z, Yu ZP, Hu HY. Up-regulated expression of matrix metalloproteinases in endothelial cells mediates platelet microvesicle-induced angiogenesis. Cell Physiol Biochem 2017;41(6):2319-2332.
Qiu F, Song J, Bi X, Wang M, Zhao Y, Fu G. Thymosin β4 promotes glucose-impaired endothelial progenitor cell function via Akt/ endothelial nitric oxide synthesis signaling pathway. Exp Ther Med 2018;16(4):3439- 3444.
Dong W, Luo B, Qiu C, Jiang X, Shen B, Zhang L, Liu W, Zhang W. TRIM3 attenuates apoptosis in Parkinson’s disease via activating PI3K/AKT signal pathway. Aging (Albany NY) 2020;13(1):735-749.
Wang Y, Lin Y, Wang L, Zhan H, Luo X, Zeng Y, Wu W, Zhang X, Wang F. TREM2 ameliorates neuroinflammatory response and cognitive impairment via PI3K/AKT/ FoxO3a signaling pathway in Alzheimer’s disease mice. Aging (Albany NY) 2020;12(20):20862-20879.
Xu L, Chen J, Jia L, Chen X, Awaleh Moumin F, Cai J. SLC1A3 promotes gastric cancer progression via the PI3K/AKT signalling pathway. J Cell Mol Med 2020 ;24(24):14392-14404.
Zhu S, Liu X, Xue M, Li Y, Cai D, Wang S, Zhang L. 20(S)-ginsenoside Rh2 induces caspase-dependent promyelocytic leukemia-retinoic acid receptor A degradation in NB4 cells via Akt/Bax/caspase9 and TNF-α/ caspase8 signaling cascades. J Ginseng Res 2021;45(2):295-304.
Zhou P, Xie W, Luo Y, Lu S, Dai Z, Wang R, Sun G, Sun X. Protective effects of total saponins of Aralia elata (Miq.) on endothelial cell injury induced by TNF-α via modulation of the PI3K/Akt and NF-κB signalling pathways. Int J Mol Sci 2018;20(1):36.
Rajendran P, Rengarajan T, Thangavel J, Nishigaki Y, Sakthisekaran D, Sethi G, Nishigaki I. The vascular endothe
lium and human diseases. Int J Biol Sci 2013;9(10):1057.
Palmer RM, Ashton D, Moncada S. Vascular endothelial cells synthesize nitric oxide from L -arginine. Nature 1988;333(6174):664- 666.
Kupatt C, Bock-Marquette I, Boekstegers P. Embryonic endothelial progenitor cell-mediated cardioprotection requires Thymosin β4. Trends Cardiovasc Med 2008;18(6):205-210.
Grant D, Kinsella J, Kibbey M, LaFlamme S, Burbelo P, Goldstein A, Kleinman, HK. Matrigel induces thymosin beta 4 gene in differentiating endothelial cells. J Cell Sci 1995;108(12):3685-3694.
Grant DS, Rose W, Yaen C, Goldstein A, Martinez J, Kleinman H. Thymosin β4 enhances endothelial cell differentiation and angiogenesis. Angiogenesis 1999;3(2):125-135.
Ho J, Su Y, Chen KH, Lee O. Protection of thymosin beta-4 on corneal endothelial cells from UVB-induced apoptosis. Chin J Physiol 2010;53(3):190-195.
Zhao Y, Qiu F, Xu S, Yu L, Fu G. Thymosin β4 activates integrin-linked kinase and decreases endothelial progenitor cells apoptosis under serum deprivation. J Cell Physiol 2011;226(11):2798-2806.
Qiu FY, Song XX, Zheng H, Zhao YB, Fu GS. Thymosin β4 induces endothelial progenitor cell migration via PI3K/Akt/eNOS signal transduction pathway. J Cardiovasc Pharmacol 2009;53(3):209-214.
Li J, Qiu F, Yu L, Zhao Y, Fu G, Zhou B. Thymosin β4 reduces senescence of endothelial progenitor cells via the PI3K/Akt/ eNOS signal transduction pathway. Mol Med Rep 2013;7(2):598-602.
Bock-Marquette I, Shrivastava S, Pipes GT, Thatcher JE, Blystone A, Shelton JM, Galindo CL, Melegh B, Srivastava D, Olson EN. Thymosin β4 mediated PKC activation is essential to initiate the embryonic coronary developmental program and epicardial progenitor cell activation in adult mice in vivo. J Mol Cell Cardiol 2009;46(5):728- 738.
Lv S, Cheng G, Zhou Y, Xu G. Thymosin beta4 induces angiogenesis through Notch signaling in endothelial cells. Mol Cell Biochem 2013;381(1):283-290.
Kim S, Kwon J. Effect of thymosin beta 4 in the presence of up-regulation of the insulinlike growth factor-1 signaling pathway on high-glucose-exposed vascular endothelial cells. Mol Cell Endocrinol 2015;401:238- 247.