益气解毒活络中药组分配伍对高糖高脂环境下人肾小球系膜细胞PPAR-γmRNA及蛋白表达的影响

Effect of Combination of Supplementing Qi, Detoxifying and Invigorating Herbal Medicines on PPAR-γ mRNA and Protein Expression in Human Glomerular Mesangial Cells under High Glucose and High-fat Condition

目的:观察益气解毒活络中药组分配伍对高糖高脂环境下人肾小球系膜细胞PPAR-γmRNA及蛋白表达影响,探讨益气解毒活络中药有效成分防治糖尿病肾病的可能机制。方法:体外培养人肾小球系膜细胞株(GMC),分为正常对照组,高糖高脂模型组、中药组(第1组、第2组、第3组),qRT-PCR法检测各组24、48 h PPAR-γmRNA的表达,WesternBlot法检测各组24、48 h PPAR-γ蛋白活性的表达。结果:(1)各组24、48 h PPAR-γmRNA表达比较:24 h:与正常组比较模型组及中药配伍组均有显著差异(P<0.01),与模型组比较各中药配伍组PPAR-γmRNA表达均明显上升,且有显著差异(P<0.01),各中药配伍组组间比较发现第1组PPAR-γmRNA表达最高(P<0.01)。48 h:与正常组比较除去第1组未见明显差异(P>0.05),模型组及其余各用药组均有显著差异(P<0.01);与模型组比较各中药配伍组PPAR-γmRNA表达均明显上升,且有显著差异(P<0.01);各中药配伍组组间比较发现第1组PPAR-γmRNA表达最高(P<0.01)。正常组24、48 h两时间点PPAR-γmRNA表达未见显著差异(P>0.05),模型组及第1、2、3组48h PPAR-γmRNA表达显著高于24 h PPAR-γmRNA表达(P<0.01)。(2)各组24、48h PPAR-γ蛋白表达比较:GMC细胞24 h PPAR-γ蛋白表达比较:模型对照组PPAR-γ蛋白低表达,各中药配伍组均能够调高PPAR-γ的表达,各中药配伍组组间比较发现第1组调高PPAR-γ作用最显著。GMC细胞48 h PPAR-γ蛋白表达比较:模型对照组PPAR-γ蛋白低表达,各中药配伍组均能够调高PPAR-γ的表达,各中药配伍组组间比较发现第1组调高PPAR-γ作用最显著。结论:益气解毒活络中药有效成分可能是通过上调PPAR-γ表达水平的作用来保护受损的GMC,从而起到减缓糖尿病肾病病程进展,保护受损肾脏的作用。

Objective:To observe the effect of combination of supplementing Qi, detoxifying and invigorating Chinese medicine on the expressions of PPAR-γ mRNA and protein in human glomerular mesangial cells under high glucose and high lipid environmentand explore the possible mechanism of effective components of supplementing Qi, detoxifying and invigorating medicine in preventing and treating diabetic nephropathy.Methods:In vitro cultured human glomerular mesangial cell lines(GMCs)were divided into normal control group, high-sugar high-fat model group, Chinese medicine groups(first group, second group, third group), and qRT-PCR method was used.The expression of PPAR-γmRNA was detected at 24 hand 48 h.The expression of PPAR-γprotein was detected by Western Blot at 24 hand 48 h.Results:(1)The expression of PPAR-γmRNA at 24 h:Compared with the normal group, there was significant difference between the model group and Chinese medicine groups(P<0.01).Compared with the model group, the expression of PPAR-γmRNA in the Chinese medicine groups was compared.All of them significantly increased and there were significant differences(P<0.01).The comparison between the groups of Chinese herbs showed that the expression of PPAR-γmRNA in the first group was the highest(P<0.01).The expression of PPAR-γmRNA at 48h: There was no significant difference between the first group and the normal group(P>0.05), and there was a significant difference between the model group and the other groups(P<0.01).Compared with the model group, the expression of PPAR-γmRNA was increased.All of them significantly increased, and there were significant differences(P<0.01).The comparison between the groups of Chinese medicine showed that the expression of PPAR-γmRNA was the highest in the first group(P<0.01).There was no significant difference in the expression of PPAR-γ mRNA between 24 h and 48 h in the normal group(P>0.05).The expression of PPAR-γmRNA in the model group and the first, second and third groups was significantly higher than that at 24 h(P<0.01).(2)Comparison of PPAR-γprotein expression at 24 h and 48 h in each group:PPAR-γ protein expression was lower in the model control group, and the expression of PPAR-γwas increased in each Chinese medicine group.The first group's PPAR-γincreased most significantly.The expression of PPAR-γprotein in GMC cells at 48 h was lower in the model control group, and the expression of PPAR-γ was increased in each Chinese medicine group.Conclusion:The active constituents of Chinese herbs for supplementing Qi, detoxifying and invigorating herbs may protect the damaged GMC by up-regulating the expression level of PPAR-γ, which can slow down the progression of diabetic nephropathy and protect the damaged kidney.