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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(P<0.01)。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(P<0.05),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。
OBJECTIVE: To observe the effects of Liuwei Dihuang Pills containing serum on the proliferation of rat mesangial cells cultured in high glucose and the expressions of monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule-1 (ICAM-1) influences. Methods: Different doses of rat serum containing Liuweidihuang Pill were prepared, and cultured in high glucose-induced rat mesangial cells. The rats were divided into two groups: control group (HG + control group), HG + control group (HG + control group), HG + low dosage group Dose group (5% Liuwei Dihuang serum + 5% control serum), HG + high dose group (10% Liuweidihuangwan serum). After treated with serum containing Liuwei Dihuang Pill for 24,48 h, the changes of mesangial cell activity and proliferation were observed by MTT method and incorporation of 5-bromodeoxyuridine (Brdu) The secretion of MCP-1 and ICAM-1 in the cell culture supernatant was measured by RT-PCR. The mRNA expressions of MCP-1 and ICAM-1 in mesangial cells were detected by RT-PCR. Results: Compared with the blank group, the proliferative activity of mesangial cells in high glucose group for 24 and 48 h, the contents of MCP-1 and ICAM-1 in the supernatant, and the expressions of MCP-1 and ICAM-1 mRNA in the high glucose group were significantly (P <0.01). Compared with the high glucose group, 5% and 10% of Liuweidihuangwan serum can reverse the above changes (P <0.05), and the inhibition was a certain time and dose-dependent. CONCLUSION: High glucose can induce mesangial cell proliferation and increase the expression of MCP-1 and ICAM-1. Liuweidihuang Pill can reduce the proliferation activity, decrease the secretion of MCP-1 and ICAM-1 and down-regulate the expressions of MCP-1 and ICAM-1 mRNA Expression, which will help delay the progress of diabetic nephropathy glomerulosclerosis.