虾青素调控SIRT1/PGC-1α/NRF1信号通路减轻对比剂诱导的急性肾小管损伤

Astaxanthin Attenuates Contrast-induced Acute Kidney Injury by Regulating SIRT1/PGC-1α/NRF1 Signaling Pathway

目的观察虾青素(astaxanthin,AST)对碘海醇(iohexol,Ⅰ)诱导的大鼠肾小管上皮细胞损伤的保护作用,并探讨其可能机制。方法常规培养的大鼠肾小管上皮细胞株(NRK-52E)分为空白对照组(Control组)、溶剂对照组(DMSO组)、碘海醇组(Ⅰ组)、虾青素预处理组(AST组)、虾青素和尼克酰胺共处理组(AST+NA组)、尼克酰胺(nicotinamide,NA组)。CCK-8检测细胞增殖情况;硫代巴比妥酸法测定丙二醛(MDA)含量;流式细胞仪检测细胞内活性氧(ROS)的水平;Western blot法检测各组细胞SIRT1、PGC-1α和NRF1蛋白表达。结果与Control组比较,碘海醇培养的NRK-52E细胞增殖活性明显下降,MDA和ROS水平升高,SIRT1、PGC-1α和NRF1蛋白表达明显下降(P <0. 05)。与Ⅰ比较,AST预处理组细胞增殖活性增加,MDA和ROS水平下降,SIRT1、PGC-1α和NRF1蛋白表达上调(P <0. 05)。在AST组基础上给予SIRT1抑制剂后,逆转了以上AST的保护作用。与AST+NA组比较,NA组细胞活性下降,MDA和ROS水平升高,SIRT1、PGC-1α和NRF1蛋白表达增加(P <0. 05)。结论虾青素通过减少MDA和ROS水平,调控SIRT1/PGC-1α/NRF1信号通路缓解碘海醇诱导的NRK-52E细胞损伤。

Objective To investigate the protective effect of astaxanthin (AST)on Rat renal tubular epithelial cells (NRK -52E) injury induced by Iohexol (I),and to explore its possible mechanism.Methods NRK -52E cells were randomly devivided into six groups :normal group (Control group),vehicle group (DMSO group),iohexol group (I group),astaxanthin group (AST group),astaxanthin plus nicotinamide group (AST +NA group),and nicotinamide group (NA group).The cell proliferation was measured by CCK-8 assay.The levels of malonaldehyde (MDA)were measured by thiobarbituric acid mthod.Intracellular reactive oxygen species (ROS) levels was measured by Flow cytometry.The protein levels of SIRT1,PGC-1α and NRF1 were detected by Western Blotting.Results Compared with Control group,I group showed suppressed proliferation,the increased levels of MDA and ROS,and the expressions of SIRT1,PGC-1α and NRF1 protein were decreased.Compared to I group,AST pretreated group presented obvious proliferation,the decreased levels of MDA and ROS,and up-expressed SIRT1,PGC-1α and NRF1 protein.SIRT1inhibitor NA administered reversed the protective effect of the AST above.Compared with the AST +NA group,NA group showed lower proliferation,the elevated levels of MDA and ROS,and the decreased expressions of SIRT1,PGC-1α and NRF1protein,which further confirmed the protective effect of AST.Conclusion Astaxanthin could alleviate iohexol -induced NRK -52E cell injury by decreasing the levels of MDA and ROS and regulating the SIRT1/PGC-1α/NRF1 signaling pathway.