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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(MTY)比色法测定24 h 和48 h 两个时间点各组细胞增殖,酶联免疫吸附试验(ELISA)测定6 h、12 h 和24 h 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)β_2:mRNA 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 LNβ_2:mRNA,LPS 组在各时间点 LNβ_2mRNA 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。
Objective To observe the effect of fosinopril (FOS) on proliferation and secretion of extracellular matrix (ECM) induced by lipopolysaccharide (LPS) in rat mesangial cells (GMCs). Methods The rat GMCs cultured in vitro were established and used for experiments after 3 to 10 generations. The experiment was divided into five groups: control group, LPS stimulation group (LPS group), Fosinopril (FOS) high, medium and low dose groups (FOS1 group, FOS2 group, FOS3 group). Methyl thiazolyl tetrazolium (MTY) colorimetric assay 24 h and 48 h two time points of each group cell proliferation, enzyme-linked immunosorbent assay (ELISA) determination of 6 h, 12 h and 24 h cell culture supernatant ECM protein content. The semi-quantitative RT-PCR was used to detect the expression of β 2: mRNA in ECM fibronectin. Results (1) LPS induced the proliferation of GMC and FOS inhibited the proliferation of GMC induced by LPS. (2) The GMC secreted a certain amount of ECM in normal cultured rats. The ECM secreted by LPS at each time point was higher than that of control group (P <0.01). (3) The normal cultured rat GMC could express a certain amount of LNβ_2: mRNA. The expression of LNβ_2 mRNA in LPS group was higher than that of LPS group In the control group, the expression of FOS in each group was lower than that in LPS group. Conclusion LPS can induce GMC proliferation and increase secretion of ECM. FOS can inhibit the proliferation of GMC induced by LPS. Inhibition of LPS-induced increase of ECM secretion induced by LPS at both protein and mRNA levels provides a theoretical basis for the protective effect of FOS on the kidney.