槲皮素对大鼠肾小管上皮细胞增殖、线粒体功能的影响及其机制研究  

The Effect of Quercetin on the Cell Proliferation and Mitochondrial Function in Rat Renal Tubular Epithelial Cells and Its Mechanism

作　　者：封宝红 伍军 李相友 喻业安 张艳霞 朱戈丽 黄徽 FENG Bao-hong;WU Jun;LI Xiang-you;YU Ye-an;ZHANG Yan-xia;ZHU Ge-li;HUANG Hui(Department of Nephrology, Wuhan Third Hospital, Wuhan 430060, China)

机构地区：[1]武汉市第三医院肾内科,武汉430060

出　　处：《微循环学杂志》2021年第3期1-6,共6页Chinese Journal of Microcirculation

基　　金：湖北省卫健委科研项目(WJ2019Q001);武汉市卫生健康科研基金(WX19C20);武汉市科技局应用基础前沿项目(2019020701011434)。

摘　　要：目的:探讨槲皮素对大鼠肾小管上皮细胞(NRK-52E)增殖、线粒体功能的影响及PI3K/AKT通路的调控作用。方法:体外培养NRK-52E细胞,将其分为空白对照组(N组)、0.4mmol/L尿酸组(U组)、0.4mmol/L尿酸+15mg/L槲皮素组(Q组),每组设6个复孔。培养24h后,倒置显微镜观察细胞的形态,MTT法测定各组细胞的增殖率,荧光探针DCFH-DA及流式细胞术检测细胞内活性氧(ROS)的生成情况,JC-1染色法于荧光显微镜下观察线粒体膜电位变化,Western blot检测磷脂酰肌醇激酶(PI3K)、蛋白激酶B(AKT)及雷帕霉素靶蛋白(mTOR)蛋白的表达。结果:与N组相比,U组NRK-52E细胞的增殖率下降(P<0.01),Q组的增殖率较U组明显上升(P<0.05)。与U组相比,Q组细胞ROS生成减少(P<0.01),线粒体膜电位增加(P<0.01)。与U组相比,Q组AKT磷酸化水平增加,PI3K、AKT、mTOR的表达亦增加(P<0.01)。结论:槲皮素对尿酸致NRK-52E细胞凋亡有保护作用,其保护作用可能是通过PI3K/AKT信号途径改善线粒体功能,从而减少细胞凋亡,促进细胞增殖来实现的。Objective:To investigate the effects of quercetin on the proliferation and mitochondrial function of rat renal tubular epithelial cells(NRK-52E)and the regulation of PI3K/AKT pathway.Method:NRK-52E cells were cultured in vitro and divided into control group(group N),0.4mmol/L uric acid group(group U),0.4mmol/L uric acid+15mg/L quercetin(group Q),each group had 6 multiple holes.After culturing for 24 hours,the cell morphology was observed with an inverted microscope,the proliferation rate of cells in each group was tested by MTT method,fluorescent probe DCFH-DA and flow cytometry were used to detect the production of reactive oxygen species(ROS)in the cells;JC-1 staining method was used to observe the changes of mitochondrial membrane potential under a fluorescence microscope,and Western blot was used to detect the expression of PI3K,AKT and mTOR proteins.Results:Compared with group N,the proliferation rate of NRK-52E cells in group U was decreased(P<0.01),and the proliferation rate of Q was significantly higher than group U.As the concentration of quercetin increased,the proliferation rate of NRK-52E cells was decreased significantly(P<0.05).The proliferation rate also showed an upward trend(P<0.01).Compared with group U,ROS production in Q group was decreased(P<0.01).Compared with group U,the phosphorylation level of AKT in group Q was increased,and the expression of PI3K,AKT,and mTOR were also increased(P<0.01).Conclusion:Quercetin has a protective effect on NRK-52E cell apoptosis induced by uric acid.Its protective effect may be achieved by improving mitochondrial function through PI3K/AKT signaling pathway,thereby reducing cell apoptosis and promoting cell proliferation.

关 键 词：槲皮素 活性氧 线粒体膜电位 

分 类 号：R589.7[医药卫生—内分泌]