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目的研究脂多糖(lipopolysaccharides,LPS)诱导大鼠肾小球系膜细胞增殖和对分泌细胞外基质的影响以及普罗布考(probucol,PRB)的保护作用。方法细胞同步化后分为:正常对照组;LPS组(LPS 10μg.mL-1);PRB+LPS处理组(PRB浓度分别为10,20,50μmol.L-1),继续培养24,48,72 h。以MTT法检测系膜细胞的增殖情况,RT-PCR法检测Ⅳ型胶原(college-Ⅳ,COL-Ⅳ)与结缔组织生长因子(connective tissue growth factor,CTGF)mRNA的表达差异,Western blot法检测COL-Ⅳ,CTGF蛋白的表达差异。结果处理24,48,72 h后,LPS组细胞增殖较对照组显著增加(P<0.05),PRB+LPS组与LPS组比较,细胞增殖显著抑制(P<0.05),并呈时间-剂量依赖效应(r=0.932,P<0.05)。培养48 h后,LPS组COL-Ⅳ,CTGF的分泌较对照组显著增加(P<0.05);预先应用不同剂量的PRB治疗后,COL-Ⅳ,CTGF mRNA和蛋白的水平较LPS组出现下调(P<0.05)。结论 LPS能促进系膜细胞增殖,诱导细胞因子的释放,促进细胞外基质的合成,参与脂毒性肾损害的发生,PRB可能通过下调这些血管生长因子的表达发挥肾保护作用。
Objective To investigate the effects of lipopolysaccharides (LPS) on the proliferation of rat mesangial cells and the secretion of extracellular matrix and the protective effect of probucol (PRB). Methods The cells were divided into three groups: normal control group, LPS group (10μg.mL-1 LPS group) and PRB + LPS group (PRB concentration 10,20,50μmol.L-1) 72 h. The proliferation of mesangial cells was detected by MTT assay. The expression of collagen Ⅳ (COL-Ⅳ) and connective tissue growth factor (CTGF) mRNA was detected by RT-PCR and Western blot COL-Ⅳ, CTGF protein expression differences. Results The proliferation of LPS group was significantly higher than that of the control group (P <0.05) 24 h, 48 h and 72 h after treatment. Compared with LPS group, the cell proliferation was significantly inhibited (P <0.05) in PRB + LPS group and in a dose-dependent manner Effect (r = 0.932, P <0.05). After cultured for 48 h, the secretion of COL-Ⅳ and CTGF in LPS group was significantly increased compared with that in control group (P <0.05). After pretreatment with different doses of PRB, the mRNA and protein expressions of COL-Ⅳ and CTGF were down-regulated P <0.05). Conclusion LPS can promote the proliferation of mesangial cells, induce the release of cytokines, promote the synthesis of extracellular matrix, and participate in the occurrence of lipotoxic renal lesion. PRB may exert renal protective effect by down-regulating the expression of these vascular growth factors.