过氧化脂质体增殖激活受体-γ对系膜细胞炎症的调控作用

Anti-inflammatory effect of peroxisome proliferator-activated receptor-γ incultured human mesangial cells

目的 了解过氧化脂质体增殖激活受体 γ(PPARγ)在核转录水平对人系膜细胞 (HMCL)炎症过程的调控作用。方法 培养人系膜细胞 ,利用IL 1β制备炎症性系膜细胞模型 ,酶联免疫吸附方法 (ELISA)测定培养细胞上清液TNF γ和IL 6水平 ;蛋白印迹方法测定HMCL中。加入不同浓度的罗格列酮 (RGZ)、曲格列酮 (TGZ)及 15d PGF2 ,PPARγ蛋白水平 ;逆转录聚合酶链反应 (RT PCR)方法检测培养细胞中PPARγ和炎症因子的mRNA表达。结果 IL 1β(10ng/ml)可使炎性细胞因子TNF γ和IL 6蛋白、mRNA表达增加 ,正常人系膜细胞可低水平表达PPARγ ,炎症刺激后PPARγ蛋白和mRNA表达显著增加 ,PPARγ特异性配基曲格列酮 (TGZ)可下调PPARγ表达。三种结构不同的PPARγ特异性配基TGZ、RGZ和 15d PGJ2 可下调炎症刺激后IL 6和TNF γ蛋白水平 ,且这种抗炎效应的作用环节在mRNA水平。结论 PPARγ可能是系膜细胞炎症与抗炎反应过程中炎症自我限制的一个重要靶位 ,作用于这一靶位的特异性配基有望成为治疗肾小球肾炎的新药。

Objective To investigate whether peroxisome proliferator-activator receptor-γ (PPARγ) could retard the inflammatory challenge from human mesangial cells. Methods Based on the inflammatory mesangial cell model followed by IL-1β, the levels of IL-6 and TNF-γ in the supernatants were measured by enzyme-linked immunosorbant assay (ELISA). PPARγ protein expression of human mesangial cell lines (HMCL) lysate was detected with Western blot. PPARγ, IL-6 and TNF-γ mRNA expression in HMCL lysate were measured by semi-quantitative RT-PCR assay. Results The levels of IL-6 and TNF-γ in HMCL supernatants were dramatically increased when cultured HMCL were stimulated by IL-1β (10 ng/ml). Protein and mRNA expressions of PPARγ in IL-1β challenge cells were significantly increased than observed in untreated cells. However, PPARγ agonist troglitazone down-regulated mRNA and protein expression of PPARγ from IL-1β challenge HMCLs. PPARγ agonists troglitazone, rosiglitazone, and 15-deoxy-Δ 12,14-prosglandin J 2 significantly decreased the upexpression of TNF-α and IL-6 in HMCL supernatants stimulated by IL-1β. Furthermore, troglitazone down-regulated TNF-α and IL-6 mRNA expression levels of HMCLs. Conclusion These data suggest that PPARγ play an important role in response to inflammatory stress in mesangial cells. PPARγ may prove to be a pharmacological target in glomerulonephritis.