两色金鸡菊提取物对大鼠肾小球系膜细胞增殖及纤维化因子表达的影响  

Effect of Coreopsis tinctoria Extract on Cell Proliferation and Fibrosis Factor Expression of High Glucose-induced Rat Glomerular Mesangial Cell

作　　者：姚蓝[1] 张雨洁[1,2] 刘菁[3] 李琳琳[3] 王烨[3] 兰怡[4] 毛新民[1] 

机构地区：[1]新疆医科大学中医学院,乌鲁木齐830011 [2]新疆医科大学第一附属医院,乌鲁木齐830054 [3]新疆医科大学基础医学院,乌鲁木齐830011 [4]新疆医科大学附属中医医院,乌鲁木齐830000

出　　处：《中国实验方剂学杂志》2017年第19期134-139,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(U1303223;81560671)

摘　　要：目的:本研究通过高糖诱导的大鼠肾小球系膜细胞建立糖尿病肾病的体外模型,观察两色金鸡菊乙酸乙酯提取物(ethyl acetate extract of Coreopsis tinctoria,AC)对模型中细胞增殖及纤维化因子表达的影响。方法:采用AC对高糖诱导大鼠肾小球系膜细胞进行干预,将细胞分为正常组,模组组,AC不同质量浓度(25,50,100,150 mg·L^(-1))组。采用噻唑蓝(MTT)比色法,细胞增殖与活性检测(CCK8)法检测AC不同质量浓度对高糖诱导大鼠肾小球系膜细胞增殖的抑制作用;采用实时荧光定量PCR(Real-time PCR),蛋白免疫印迹法(Western blot)进一步观察AC不同质量浓度对肾脏纤维化蛋白转化生长因子-β_1(TGF-β_1),胶原蛋白Ⅳ(CollagenⅣ),纤维连接蛋白(FN)表达的影响。结果:AC 50,100,150 mg·L^(-1)组均能抑制高糖诱导大鼠肾小球系膜细胞的增殖(P<0.01);AC 25,100,150 mg·L^(-1)质量浓度组显著降低细胞中TGF-β_1的mRNA与蛋白表达水平(P<0.01);AC 100,150 mg·L^(-1)质量浓度组显著抑制细胞中CollagenⅣmRNA与蛋白的表达水平(P<0.01);AC 50,100,150 mg·L^(-1)质量浓度组显著抑制细胞中FN mRNA与蛋白的表达水平(P<0.01)。结论:两色金鸡菊乙酸乙酯提取物可能通过抑制大鼠肾小球系膜细胞增殖及纤维化因子表达发挥其对糖尿病肾病肾脏保护作用。Objective： To establish the vitro diabetic nephropathy model with high-glucose induced-rat glomerular mesangial cells to detect the inhibitory effect of ethyl acetate extract of Coreopsis tinctoria （AC） on cell proliferation and fibrosis factor expression. Method： High glucose-induced rat glomerular mesangial cells were treated with AC. The cells were divided into six groups：normal group, high-glucose-induced model group, high-glucose＋AC groups, with concentrations of 25, 50,100,150 mg·L^-1. MTT and cell counting kit 8 （CCK8） were used to test the inhibitory effect of AC on cell proliferation. Real-time PCR and Western blot were used to test the inhibitory effect of AC on transforming growth factor-β1 （TGF-β1）, Collagen Ⅳ, fibronectin （FN） mRNA and protein expressions. Result： AC with concentrations of 25, 50, 100, 150 mg·L^-1 significantly inhibited the proliferation of glomerular mesangial cells of high-glucose-induced rats （P〈0.01）. AC with concentrations of 25, 100, 150 mg·L^-1 significantly inhibited TGF-β1 mRNA and protein expressions （P〈0.01）. AC with concentrations of 100, 150 mg·L-1 significantly inhibited Collagen IV mRNA and protein expressions （P〈0.01）. AC with concentrations of 50, 100, 150 mg·L^-1 significantly inhibited FN mRNA and protein expressions （P〈0.01）. Conclusion： AC prevents the diabetic nephropathy renal fibrosis by suppressing proliferation of rat glomerular mesangial cells and fibrosis factor expression.

关 键 词：两色金鸡菊乙酸乙酯提取物 大鼠肾小球系膜细胞 细胞增殖 纤维化因子表达 

分 类 号：R966[医药卫生—药理学] R285.5[医药卫生—药学]