Abstract
Testosterone deficiency is associated with a higher incidence of cardiovascular diseases in men. However, its effect on cell senescence, which plays a causal role in vascular aging, remains unclear. Here, we tested the hypothesis that testosterone alleviated vascular smooth muscle cell (VSMC) senescence and collagen synthesis via growth arrest-specific protein 6 (Gas6)/Axl- and Akt/FoxO1a-dependent pathways. Testosterone significantly ameliorated angiotensin II-induced VSMC senescence and collagen overexpression. In addition, testosterone inhibited angiotensin II-induced matrix metalloproteinase-2 (MMP-2) activity, which played a pivotal role in facilitating age-related collagen deposition. Testosterone increased the expression of tissue inhibitor of metalloproteinase-2 but decreased the expression of MMP-2 and membrane type-1 metalloproteinase which contributed to increase MMP-2 activity. The effects on VSMCs senescence and collagen synthesis were mediated by restoration of angiotensin II-induced downregulation of Gas6 and Axl expression and a subsequent reduction of Akt and FoxO1a phosphorylation. The effects of testosterone were reversed by a Gas6 blocker, Axl-Fc, and a specific inhibitor of Axl, R428. Treatment of VSMCs with PI3K inhibitor LY294002 abrogated the downregulating effect of testosterone on MMP-2 activity. Furthermore, when FoxO1a expression was silenced by using a specific siRNA, the inhibitory effect of testosterone on MMP-2 activity was revered as well, that indicated this process was Akt/FoxO1a dependence. Taken together, Gas6/Axl and Akt/FoxO1a were involved in protective effects of testosterone on VSMCs senescence and collagen synthesis. Our results provide a novel mechanism underlying the protective effect of testosterone on vascular aging and may serve as a theoretical basis for testosterone replacement therapy.
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Acknowledgments
This work was supported by the research grants from the National Basic Research Program of China (973 Program, Grant No. 2013CB530700), the National Natural Science Foundation of China (81100605, 81270352, 81270287, 81300168, 81471036, 81470560, and 81570400), the Natural Science Foundation of Shandong Province (BS2013YY017, ZR2014HQ037), the Key Research and Development Program of Shandong Province (2015GSF118062), cardiovascular exploration research foundation of Chinese Medical Doctor Association (DFCMDA201320), and the Specialized Research Fund for the Doctoral Program of Higher Education (SRFDP 20130131120065).
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The Akt signaling pathway is involved in the anti-senescence effect of testosterone (A) The PI3K inhibitor, LY294002 could reverse the downregulating effects of testosterone on the expression of p16INK4a and p21Cip1 (P<0.05) induced by angiotensin II. But the regulating effect of LY294002 on p16INK4a expression was much weaker. Angiotensin II (Ang II), testosterone (T). Values are mean±SD of three measurements. *P<0.05 and **P<0.01 compared with control group; #P<0.05 and ##P<0.01compared with angiotensin II-treated group; †P<0.05 compared with testosterone-treated group (GIF 54 kb)
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Chen, Yq., Zhao, J., Jin, Cw. et al. Testosterone delays vascular smooth muscle cell senescence and inhibits collagen synthesis via the Gas6/Axl signaling pathway. AGE 38, 60 (2016). https://doi.org/10.1007/s11357-016-9910-5
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DOI: https://doi.org/10.1007/s11357-016-9910-5