福辛普利对大鼠肾小球系膜细胞增殖及分泌细胞外基质的影响

目的观察福辛普利（FOS）对脂多糖（LPS）诱导的大鼠肾小球系膜细胞（GMC）增殖及分泌细胞外基质（ECM）的影响。方法建立体外培养的大鼠GMC,鉴定后3—10代用于实验。实验分为五组：对照组、LPS刺激组（LPS组）、福辛普利（FOS）高、中、低剂量组（分别为FOS1组、FOS2组、FOS3组）。甲基噻唑基四唑（MTT）比色法测定24h和48h两个时间点各组细胞增殖,酶联免疫吸附试验（ELISA）测定6h、12h和24h细胞培养上清液中ECM蛋白含量;荧光半定量RT-PCR法检测ECM纤维连接蛋白（LN）β2mRNA表达的变化。结果（1）LPS可诱导GMC增殖,FOS可抑制LPS诱导的GMC增殖;（2）正常培养的大鼠GMC可分泌一定量的ECM,LPS组在各时间点分泌ECM均高于对照组（P〈0．01）,而FOS各组分泌ECM均低于LPS组（P〈0．01）;（3）正常培养的大鼠GMC可表达一定量LNβ2 mRNA,LPS组在各时间点LNβ2mRNA表达量均高于对照组,FOS各组表达量均低于LPS组。结论LPS可诱导GMC增殖且分泌ECM增加,FOS可抑制LPS诱导的GMC增殖,从蛋白和mRNA两个水平抑制LPS诱导的GMC分泌ECM增加,为FOS对肾脏的保护作用提供了理论依据。

Effects of fosinopril on proliferation and secretion of extracellular matrix of rat glomerular mesangial cell

OBJECTIVE: To observe the effects of fosinopril (FOS) on proliferation and secretion of extracellular matrix of rat glomerular mesangial cell induced by LPS. METHODS: In vitro culture method for glomerular mesangial cells (GMC) of rat was established and passages 3 - 10 of the cells were used in the experiment after identification. The experiment included the following 5 groups: control group (Ctrl), LPS group (LPS), high, medium and low dose FOS groups (FOS1, FOS2 and FOS3 groups, respectively). GMC proliferation was detected by methyl thiazolyl tetrazolium (MTT) incorporation method at 24 and 48 h; the changes of laminin (LN), fibronectin (FN) and ColIV protein secretion was detected by the enzyme-linked immunosorbent assay (ELISA). The changes of LNbeta(2) mRNA expression was detected by semi-quantitative real-time RT-PCR. RESULTS: (1) LPS could induce the mesangial cell proliferation, FOS inhibited this effect of proliferation induced by LPS. (2) Mesangial cells could secrete some extracellular matrix (ECM) protein in normal culture medium, mesangial cell secreted ECM protein was significantly higher in LPS group than that in Ctrl group (P < 0.01), but significantly lower in all FOS groups than that in LPS group (P < 0.01). (3) Mesangial cell could express LNbeta(2) mRNA in normal culture medium, LNbeta(2) mRNA expression was significantly higher in LPS group than that in Ctrl group at all time points, but was significantly lower in FOS group than that in LPS group. CONCLUSIONS: LPS could induce increased secretion of the ECM, including LN, FN, ColIV; FOS could inhibit the secretion of ECM in GMC in a dose-dependent manner at mRNA and protein levels.