虾青素通过SIRT1/FOXO3a信号通路预防低渗性对比剂诱导的急性肾损伤的实验研究  

Experimental Study of Astaxanthin on Contrast Agent-induced Acute Kidney Injury Through the SIRT1/FOXO3 a Signal Pathway

作　　者：刘娜娜 李文华 张德兵 郑迪 张权 

机构地区：[1]徐州医科大学心血管病研究所,221001 [2]徐州医科大学附属医院心内科

出　　处：《医学研究杂志》2017年第12期68-72,共5页Journal of Medical Research

基　　金：江苏省六大人才高峰基金资助项目(2014-YY-007)

摘　　要：目的探讨虾青素(astaxanthin,AST)通过沉默信息调节因子(Sir2)家族成员之一的SIRT1/叉头框蛋白(FOXO3a)对碘海醇诱导的人肾小管上皮(HK-2)细胞氧化损伤的作用。方法 HK-2细胞株培养贴壁后随机分为对照组、二甲基亚砜(DMSO)对照组、碘海醇组、碘海醇+SIRT1抑制剂尼克酰胺(niacinamide,NA)组、碘海醇+FOXO3a si RNA组、碘海醇+低、高剂量虾青素(1.0、10.0μmol/L)组,继续培养然后应用细胞计数试剂盒(CCK-8)检测细胞存活率,流式细胞仪检测细胞内活性氧(ROS)的水平,蛋白免疫印迹法检测细胞中SIRT1及FOXO3a的蛋白水平。结果与对照组相比,碘海醇组、碘海醇+低、高剂量虾青素组细胞存活率降低,并且随着碘海醇剂量的增加,抑制作用增强;虾青素组(在一定浓度范围内)可改善碘海醇对HK-2细胞增殖的抑制作用,一定范围内剂量增加,改善能力越强。加入碘海醇后,细胞内ROS水平明显增加,虾青素预处理后,ROS水平下调。碘海醇使SIRT1蛋白的表达上调,同时降低了叉头框蛋白(FOXO3a)的表达,虾青素可以拮抗这一作用,在一定浓度范围内,随着浓度增加,拮抗作用增强。碘海醇作用下,SIRT1抑制剂NA可以降低SIRT1的表达,增加叉头框蛋白的表达,FOXO3 a si RNA使FOXO3 a的表达下降,但对SIRT1的表达无明显影响。结论虾青素可以对抗碘海醇诱导的HK-2的氧化损伤,其可能的机制与降低内皮细胞SIRT1的表达有关,且在FOXO3a的上游发挥对抗碘海醇诱导的氧化损伤作用。Objective To explore the effect of astaxanthin（AST）on iohexol-induced injury in Human renal tubular epithelial cells （HK-2）by SIRT1/FOXO3a pathway with the possible mechanisms.Methods HK-2 cells were cultured in DMEM/F12 medium in vitro and then randomly divided into appropriate experimental groups：normal group, dimethyl sulfoxide （DMSO）, iohexol group, iohexol ＋ （1.0, 10.0μ mol/L） astaxanthin group, iohexol ＋ SIRT1 inhibitors （Nicotinamide, NA） group and iohexol ＋ FOXO3a siRNA group.When cultured for 24 hours, cell proliferation ability was tested by CCK-8 method, ROS were detected by flow cytometry. The expression of SIRT1 and FOXO3a were observed using Western blot.Results Compared with normal group, iohexol group reduced the proliferation of cells in each group,and with the increase of concentration, inhibition increased. Astaxanthin groups within a certain range of concentrations improved HK-2 cells proliferation, and as the dose increases, anti-inhibition increased.After adding iohexol, intracellular ROS levels were significantly increased,and after pretreatmented with astaxanthin, ROS significantly reduced.Iohexol upregulated SIRTl expression and reduced forkhead box protein （FOXO3a） expression. Astaxanthin can antagonize this effect. In a certain concentration range, with increasing concentration, antagonism increased.4Under iohexol, SIRT1 inhibitors nicotinamide （NA） can reduce the expression of SIRT1 and increase the expression of forkhead box protein. FOXO3a siRNA reduces the expression of FOXO3a, but had no significant effect on the expression of SIRT1.Conclusion Astaxanthin inhibits iohexol induced damage in HK-2, and its possible mechanism is related to decreasing expression of endothelial cells SIRT1,which on the upstream of FOXO3a, plays against damage induced by iohexol.

关 键 词：对比剂急性肾损伤 虾青素 沉默信息调节因子 叉头框蛋白 

分 类 号：R5[医药卫生—内科学]