苦参碱对脂多糖诱导的大鼠系膜细胞增殖和分泌细胞外基质的影响

Effects of Matrine on glomerular mesangial cell proliferation and extracellular matrix production in rat induced by LPS

目的了解不同浓度苦参碱(Matrine,Ma)对脂多糖(LPS)诱导的大鼠系膜细胞(MC)增殖及分泌细胞外基质(ECM)的影响,探讨Ma有无抗肾小球硬化的药理作用。方法采用体外培养MC,分为空白组、LPS组及Ma处理组(LPS10μg/ml+Ma20、40、80、160、320、640μg/ml),继续培养24、48、72h,以四甲基偶氮唑蓝法(MTT)检测MC的增殖情况,以台盘蓝法检测Ma对MC的细胞毒性作用,以放射免疫分析法(RIA)检测MC分泌型胶原(Col"Ⅳ)与层粘连蛋白(LN)的情况。结果经Ma处理24、48、72h后,LPS组MC增殖较空白组显著增加(P<0.01),Ma组与LPS组比较,MC增殖呈显著抑制(P均<0.01),呈时间-剂量依赖效应(r=0.826,P<0.01)。Ma在20～640μg/ml浓度间对MC无明显细胞毒作用。Ma处理48h后,LPS组Col"Ⅳ与LN的分泌较空白组显著增加(P均<0.01);Ma组对Col"Ⅳ与LN分泌的抑制作用较LPS组显著(P均<0.01),并呈剂量依赖效应(r=-0.794,-0.851,P均<0.01)。结论Ma可明显抑制LPS诱导的MC增殖和分泌细胞外基质,无明显细胞毒作用,并呈剂量依赖效应,初步认为Ma在细胞水平上有抗肾小球硬化的药理作用。

Objectives To determine the effects of different concentrations of Matrine （Ma） on glomerular mesangial cell （MC） proliferation and secretion of extracellular matrix （ECM） in rat induced by lipopolysaccharide （LPS）, and to expl.ore the protective roles of Ma against glomerul.oscl.erosis. Methods Different doses of Ma （20, 40, 80, 160, 320, 640 μg,/ml） were added to cultured MC which were exposed to LPS for 24, 48 and 72 hours. The inhibitory effect and cell toxicity of Ma on MC were analyzed by methyl thiazolyl tetrazolium （MTT） assay and cell counting. The cuhnred cells were divided into 8 groups： control, LPS group （10 μg/ml） and Ma group （LPS 10 μg/ml ＋20, 40, 80, 160, 320, 640 μg,/ml） . After 48 hours, secretion of type Ⅳ collagen （Col-Ⅳ ） and laminin （LN） were detected by radioimmunoassay. Results Compared with the control, group, MC proliferation of the LPS group significantly increased with increased exposure to Ma from 24 - 72 hours. The inhibitory effect on MC proliferation in the Ma group was significantly increased compared with that of the LPS group and was positively correlated with the dose of Ma （P 〈 0.01, r = 0.826）. No cell toxicity was observed when Ma was applied at the concentrations of 20 - 640 μg/ml. After 48 hours of exposure to Ma, the amounts of Col-Ⅳ and LN secreted in the LPS group were significantly higher than that in the Ma and the control groups. A dose-dependent correlation existed between Ma and the secretion of Col-Ⅳ and LN （r = -0.851, r = -0.794） . Conclusions Matrine inhibited glomerular MC proliferation and ECM production induced by LPS in a dose-dependent manner with no obvious cell toxicity. Matrine might have a protective role against glomerulosclerosis.