福辛普利对大鼠肾小球系膜细胞增殖及转化生长因子β_1表达的影响

Effects of Fosinopril on Expression of Transforming Growth Factor β_1 of Rat Glomerular Mesangial Cell

目的观察新型血管紧张素转化酶抑制剂（angiotension—converting enzyme inhibitor，ACED——福辛普利（fosinopril，FOS）对脂多糖（lipopolysaccharide，LPS）诱导的大鼠肾小球系膜细胞（glomerular mesangial cell，GMC）增殖及转化生长因子（transforming growth factor，TGF）β1表达的影响。方法建立体外培养大鼠肾小球系膜细胞，鉴定后3～10代用于实验。实验分为5组：对照组（Ctrl组）、脂多糖刺激组（LPS组）、福辛普利高、中、低剂量组（分别为FOS1组、FOS2组和FOS3组），分别于24h和48h两个时间点甲基噻唑基四唑（methyl thiazolyl tetrazolium，MTT）比色法测定各组细胞增殖情况；于6h，12h，24h后酶联免疫测定（enzyme—linked immunosorbent—assay，ELISA）法测定细胞培养上清液中转化生长因子β1蛋白的表达量，荧光半定量RT～PCR法检测转化生长因子β1mRNA表达的变化。结果脂多糖可诱导大鼠肾小球系膜细胞增殖，福辛普利可抑制脂多糖诱导的大鼠肾小球系膜细胞增殖；正常培养的大鼠肾小球系膜细胞均有一定量的转化生长因子β1蛋白分泌和mRNA表达；LPS组在各时间点转化生长因子β1蛋白分泌和mRNA表达均高于Ctrl组（P〈0．01），而FOS1组、FOS2组、FOS3组转化生长因子β1蛋白分泌和mRNA表达均低于LPS组（P〈0．01）。结论脂多糖可诱导大鼠肾小球系膜细胞增殖。福辛普利可抑制脂多糖诱导的大鼠肾小球系膜细胞增殖。福辛普利能够从蛋白水平和mRNA水平抑制脂多糖诱导的大鼠肾小球系膜细胞转化生长因子β1的表达，为福辛普利的肾脏保护作用提供理论依据。

Objective To observe effects of fosinopril （FOS） on proliferation and expression of transforming growth factor （TGF） β1 of rat glomerular mesangial cell （GMC） induced by lipopolysaccharide（LPS）. Methods After establishing the cultured glomerular mesangial cells of rat in vitro, passages 3-10 of cells were used in the experiment after identification. The experiment included 5 groups： control group （Ctrl group）, lipopolysaccharide group （LPS group）, high dose fosinopril group （FOS 1 group）, middle dose fosinopril group（FOS 2 group） and low dose fosinopril group（FOS 3 group）. Glomerular mesangial cells proliferation were detected by methyl thiazolyl tetrazolium （MTT） incorporation method at 24 h and 48 h, and the change of transforming growth factor β1 protein secretion was detected by the enzyme-linked immunosorbent-assay （ELISA）. The change of transforming growth factor β1 mRNA expression was detected by semi-quantitative real-time （RT）-PCR. Results Lipopolysaccharide can induce the glomerular mesangial cell proliferation, fosinopril can inhibit the effect of proliferation induced by lipopolysaccharide. Gomerular mesangial cells had transforming growth factor β1 protein secretion and mRNA expression at certain level in normal culture medium, transforming growth factor β1 protein secretion and mRNA expression were higher significantly in LPS group than those in Ctrl group （P〈0. 01）, but lower significantly in all of fosinopril groups than those in LPS group （P〈0.01）. Conclusion Fosinopril can inhibit the effect of proliferation and the expression of transforming growth factor β1 in glomerular mesangial cells at the mRNA and protein levels induced by lipopolysaccharide, and provides the theoretical evidence for the renal protection of fosinopril.