维生素D通过调节MAPK/SirT1轴保护内皮细胞免受辐射诱导的衰老和凋亡。
Vitamin D protects endothelial cells from irradiation-induced senescence and apoptosis by modulating MAPK/SirT1 axis.
作者信息
Marampon F, Gravina G L, Festuccia C, Popov V M, Colapietro A, Sanità P, Musio D, De Felice F, Lenzi A, Jannini E A, Di Cesare E, Tombolini V
机构信息
Division of Radiotherapy and Radiobiology, Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, L'Aquila, Italy.
Department of Radiation Oncology, University of Pennsylvania, Philadelphia, PA, USA.
出版信息
J Endocrinol Invest. 2016 Apr;39(4):411-22. doi: 10.1007/s40618-015-0381-9. Epub 2015 Sep 3.
PURPOSE
Radiotherapy toxicity is related to oxidative stress-mediated endothelial dysfunction. Here, we investigated on radioprotective properties of Vitamin D (Vit.D) on human endothelial cells (HUVEC).
METHODS
HUVEC, pre-treated with Vit.D, were exposed to ionizing radiation (IR): ROS production, cellular viability, apoptosis, senescence and western blot for protein detection were performed. The role of MAPKs pathway was investigated by using U0126 (10 μM) MEKs/ERKs-, SB203580 (2.5 μM) p38-inhibitor or by over/expressing MKK6 p38-upstream activator.
RESULTS
Vit.D reduced IR-induced ROS production protecting proliferating and quiescent HUVEC from cellular apoptosis or senescence, respectively, by regulating MAPKs pathways. In proliferating HUVEC, Vit.D prevented IR-induced apoptosis by activating ERKs while in quiescent HUVEC counteracted IR-induced senescence by inhibiting the p38-IR-induced activation. MEKs&ERKs inhibition in proliferating or MKK6/mediated p38 activation in quiescent HUVEC, respectively, reverted anti-apoptotic or anti-senescent Vit.D properties. SirT1 protein expression levels were up-regulated by Vit.D. ERKs inhibition blocked Vit.D-induced SirT1 protein up-regulation in proliferating cells. In quiescent HUVEC cells, p38 inhibition counteracted the IR-induced SirT1 protein down-regulation, while MKK6 transfection abrogated the Vit.D positive effects on SirT1 protein levels after irradiation. SirT1 inhibition by sirtinol blocked the Vit.D radioprotective effects.
CONCLUSION
Vit.D protects HUVEC from IR induced/oxidative stress by positively regulating the MAPKs/SirT1 axis.
目的
放射治疗毒性与氧化应激介导的内皮功能障碍有关。在此，我们研究了维生素D（Vit.D）对人内皮细胞（HUVEC）的辐射防护特性。
方法
用Vit.D预处理HUVEC，然后使其暴露于电离辐射（IR）：检测活性氧（ROS）生成、细胞活力、凋亡、衰老情况，并进行蛋白质检测的蛋白质印迹分析。通过使用U0126（10μM）抑制MEKs/ERKs、SB203580（2.5μM）抑制p38或通过过表达/表达MKK6（p38上游激活剂）来研究丝裂原活化蛋白激酶（MAPKs）途径的作用。
结果
Vit.D通过调节MAPKs途径减少IR诱导的ROS生成，分别保护增殖期和静止期的HUVEC免于细胞凋亡或衰老。在增殖期HUVEC中，Vit.D通过激活ERK来防止IR诱导的凋亡，而在静止期HUVEC中，通过抑制p38-IR诱导的激活来抵消IR诱导的衰老。分别抑制增殖期HUVEC中的MEKs&ERKs或静止期HUVEC中MKK6介导的p38激活，可逆转Vit.D的抗凋亡或抗衰老特性。Vit.D上调沉默调节蛋白1（SirT1）蛋白表达水平。抑制ERK可阻断Vit.D诱导的增殖期细胞中SirT1蛋白上调。在静止期HUVEC细胞中，抑制p38可抵消IR诱导的SirT1蛋白下调，而转染MKK6可消除照射后Vit.D对SirT1蛋白水平的积极影响。用sirtinol抑制SirT1可阻断Vit.D的辐射防护作用。
结论
Vit.D通过正向调节MAPKs/SirT1轴保护HUVEC免受IR诱导的氧化应激。
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