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目的:研究雷公藤内酯对血管内皮细胞生长因子(VEGF)mRNA表达及VEGF合成与分泌的影响,进一步探讨雷公藤内酯降低肾小球肾炎患者尿蛋白的作用机制。方法:以人内皮细胞系ECV-304为研究对象,利用半定量逆转录聚合酶链反应(RT-PCR),流式细胞仪,酶联免疫吸附法(ELISA)检测不同剂量雷公藤内酯对佛波脂(TPA)诱导的内皮细胞VEGFmRNA表达及VEGF合成与分泌的影响,用RT-PCR检测雷公藤内酯对内皮细胞c-fos/c-jun mRNA表达的影响。结果:TPA能够明显上调VEGF mRNA表达,蛋白合成与分泌。而雷公藤内酯可以抑制TPA诱导的内皮细胞VEGF mRNA表达及VEGF蛋白合成与分泌,该作用在10μg·L~(-1)时更为明显。同样,雷公藤内酯剂量依赖性地抑制TPA诱导的内皮细胞c-fos/c-jun mRNA的表达。结论:雷公藤内酯通过影响c-fos/c-jun基因转录而抑制内皮细胞VEGFmRNA表达及VEGF合成与分泌是雷公藤内酯降低肾小球肾炎患者尿蛋白的作用机制之一。
Objective: To study the effects of triptolide on the expression of vascular endothelial growth factor (VEGF) mRNA and the synthesis and secretion of VEGF, and further explore the mechanism of action of triptolide on the reduction of urinary protein in patients with glomerulonephritis. METHODS: Human endothelial cell line ECV-304 was used as the research object. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), flow cytometry, and enzyme-linked immunosorbent assay (ELISA) were used to detect different doses of triptolide against Buddha. The effect of triptolide on the expression of c-fos/c-jun mRNA in endothelial cells was detected by RT-PCR. Results: TPA could upregulate the expression of VEGF mRNA, protein synthesis and secretion. Triptolide can inhibit TPA-induced endothelial cell VEGF mRNA expression and VEGF protein synthesis and secretion, the effect is more obvious at 10μg · L -1. Similarly, triptolide inhibited TPA-induced endothelial cell c-fos/c-jun mRNA expression in a dose-dependent manner. CONCLUSION: Triptolide inhibits the transcription of c-fos/c-jun gene and inhibits the expression of VEGFmRNA and the synthesis and secretion of VEGF in endothelial cells. Triptolide is one of the mechanisms that reduce the urine protein in patients with glomerulonephritis.