六味地黄丸含药血清减弱高糖环境下肾系膜细胞的增殖和炎性因子表达

目的:观察六味地黄丸含药血清对高糖环境下培养的大鼠系膜细胞增殖和单核细胞趋化蛋白-1(MCP-1)和细胞间黏附分子-1(ICAM-1)表达的影响。方法:制备不同剂量大鼠六味地黄丸含药血清,并培养高糖诱导的大鼠系膜细胞。分别给予以下分组处理:空白(NG)组,高糖(HG)组,HG+对照血清组(10%对照血清),HG+低剂量组(2.5%六味地黄丸血清+7.5%对照血清),HG+中剂量组(5%六味地黄丸血清+5%对照血清),HG+高剂量组(10%六味地黄丸血清)。六味地黄丸含药血清处理24,48 h后,采用四甲基偶氮唑盐法和5-溴脱氧尿嘧啶核苷(Brdu)掺入法观察系膜细胞活性和增殖的变化,使用ELISA检测细胞培养上清液中MCP-1和ICAM-1分泌量,RT-PCR法检测系膜细胞MCP-1和ICAM-1 mRNA表达。结果:与空白组比较,高糖组在培养的24,48 h内系膜细胞的增殖活性、上清液中MCP-1和ICAM-1的含量,MCP-1和ICAM-1 mRNA的表达显著上调(P0.01)。与高糖组比较,5%和10%六味地黄丸含药血清均能逆转上述变化(P0.05),且抑制作用呈一定的时间、剂量依赖关系。结论:高糖可致系膜细胞增殖活性和MCP-1和ICAM-1的表达上调,六味地黄丸可降低增殖活性,减少MCP-1和ICAM-1分泌,下调MCP-1和ICAM-1 mRNA表达,从而有利于延缓糖尿病肾病肾小球硬化的进展。

Liuwei Dihuang Wan Drug Serum Attenuates Proliferation of Mesangial Cells and Inflammatory Cytokines Expression Induced by High Glucose

Objective: To observe the effect of Liuwei Dihuang Wan drug serum on the cells proliferation and expression of monocyte chemoattractant protein-1(MCP-1) and intercellular adhesion molecule excluded-1(ICAM-1) in mesangial cells of rats cultured in high glucose conditions. Method: Different doses of Liuwei Dihuang Wan drug serum were prepared, and rat mesangial cells were cultured under high glucose. grouping:normal group(NG), high glucose(HG) group, HG+control serum group(10% control serum),HG+low dose group(2.5% Liuwei Dihuang Wan serum+7.5% control serum), HG+middle dose group(5% Liuwei Dihuang Wan serum+5% control serum),HG+high dose group(10% Liuwei Dihuang Wan serum). Following treatment with Liuwei Dihuang Wan drug serum for 24, 48 h, mesangial cells activity and proliferation changes were observed with methyl thiazolyl tetrazolium and Brdu incorporation assay. MCP-1 and ICAM-1 secretion in cell culture supernatants were detected by ELISA. MCP-1 and ICAM-1 mRNA expression were measured by Real Time-PCR. Result: Compared with normal group, the cells proliferative activity, MCP-I and ICAM-1 content in supernatant, and the expression of MCP-1 and ICAM-1 mRNA were significantly up-regulated in high glucose group after 24,48 h culture(P<0.01). Compared with the high glucose group, treatment with 5% or 10% Liuwei Dihuang Wan drug serum significantly reversed these above changes(P<0.05), and the inhibition was in a time and dose-dependent manner. Conclusion: High glucose can induce up-regulation of cells proliferation, MCP-1 and ICAM-1 expression in rat mesangial cells. Liuwei Dihuang Wan may reduce the proliferation activity, decrease MCP-1 and ICAM 1-secretion, and down-regulate MCP-1 and ICAM-1 mRNA expression, thus delaying glomerulosclerosis in diabetic nephropathy.