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目的 探讨大黄酸对转化生长因子β1(TGF-β1)诱导人脐静脉内皮细胞纤溶酶原激活物抑制物1(PAI-1)mRNA表达和蛋白合成的影响,以阐明大黄酸对内皮细胞的保护作用。方法 以人脐静脉细胞系ECV-304为研究对象,Northern blot检测PAI-1 mRNA的表达,流式细胞仪法检测PAI-1蛋白的合成,免疫沉淀法和Western blot检测p44/p42丝裂原活化蛋白激酶(MAPK)活性。结果 大黄酸可以快速抑制TGF-β1诱导的内皮细胞PAI-1 mRNA的表达,且呈明显的剂量依赖效应,如果作用时间延长,则对蛋白的合成同样具有明显抑制作用。进一步的研究还发现,大黄酸对TGF-β1激活的p44/p42 MAPK活性具有明显下调作用。结论 大黄酸对TGF-β1诱导的PAI-1高表达的抑制作用可能是其保护内皮细胞功能从而防治糖尿病及其血管并发症的作用机制之一。
Objective To investigate the effect of rhein on the mRNA and protein synthesis of PAI-1 induced by transforming growth factor-β1 (TGF-β1) in human umbilical vein endothelial cells Protective effects. Methods The human umbilical vein cell line ECV-304 was used as the research object. The expression of PAI-1 mRNA was detected by Northern blot. The PAI-1 protein was detected by flow cytometry. The p44 / p42 mitogen Activated protein kinase (MAPK) activity. Results Rhein can rapidly inhibit the expression of PAI-1 mRNA induced by TGF-β1 in a dose-dependent manner. If prolonged, rhein can inhibit the synthesis of PAI-1 mRNA. Further studies also found that rhein has a significant down-regulation of p44 / p42 MAPK activity by TGF-β1. Conclusion Rhein inhibits the high expression of PAI-1 induced by TGF-β1, which may be one of the mechanisms that protects the function of endothelial cells to prevent and treat diabetes mellitus and its vascular complications.