普罗布考对脂多糖诱导的大鼠肾小球系膜细胞增殖和分泌细胞外基质的影响

Effects of Probucol on Mesangial Cell Proliferation and Extracellular Matrix Production in Rat Induced by Lipopolysaccharides

关键词:  普罗布考  脂多糖  结缔组织生长因子  胶原-Ⅳ

Key words:  probucol  lipopolysaccharides  connective tissue growth factor  college-Ⅳ

摘要:

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 10 μg·mL^-1)；PRB+LPS处理组(PRB浓度分别为10，20，50 μmol·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。

Abstract:

OBJECTIVE To study lipopolysaccharides(LPS) induced mesangial cell proliferation and secretion of extracellular matrix, and the protective effect of probucol(PRB). METHODS After synchronization, cells were divided into: control group; LPS group(LPS: 10 μg·mL^-1); PRB+LPS treatment group(PRB: 10, 20, 50 μmol·L^-1). Continued to train 24, 48, 72 h. The proliferation of mesangial cells were detected by MTT. College-Ⅳ(COL-Ⅳ), connective tissue growth factor(CTGF) mRNA expression were detected by RT-PCR. COL-Ⅳ, CTGF protein expression were detected by Western blot assay. RESULTS After treatment 24, 48, 72 h, cell proliferation of LPS group compared with control group increased significantly(P<0.05); compared with LPS group, PRB+LPS groups cell proliferation were significantly inhibited(P<0.05), and were time-dose- dependent effect(r=0.932, P<0.05). After cultivated 48 h, COL-Ⅳ, CTGF secretion of LPS group increased significantly(P<0.05); pretreatment with different doses of PRB, COL-Ⅳ, CTGF mRNA and protein levels compared with LPS group were lower(P<0.05). CONCLUSION LPS can increase mesangial cell proliferation and the release of cytokines, to promote the synthesis of extracellular matrix, involved in the occurrence of renal toxicity lipid. PRB may be attributed to the expression of these cell growth factors and play a role in kidney protection.