川芎嗪对高糖诱导后人腹膜间皮细胞血管内皮生长因子表达的影响

Effect of ligustrazine on high glucose-induced VEGF expression in human peritoneal mesothelial cells

目的研究川芎嗪对体外培养的人腹膜间皮细胞在高糖刺激下血管内皮生长因子(VEGF)表达的影响。方法体外培养人腹膜间皮细胞,分为5组(每组设3个样本):正常对照组(完全培养液)、高糖对照组(2.5%葡萄糖)和高糖(2.5%葡萄糖)加川芎嗪(浓度依次为10,20,40 mg/L)组。采用半定量逆转录聚合酶链反应(RT-PCR)法检测细胞内VEGF mRNA的表达;酶联免疫吸附测定法(ELISA)检测细胞上清液中VEGF的蛋白质水平,用二喹啉甲酸蛋白检测法测定细胞中蛋白质含量,用以校正ELISA结果。结果川芎嗪能显著降低高糖所致的人腹膜间皮细胞VEGF的表达(P<0.01);两者在蛋白质和基因水平均呈量效关系。结论川芎嗪可以减少高糖刺激下人腹膜间皮细胞VEGF的表达,从而达到延缓腹透相关性腹膜纤维化的发生和发展。

Purpose To investigate the effects of ligustrazine on the expression of vascular endothelial growth factor（VEGF）in cultured human peritoneal mesothefial cells（HPMCs）under high glucose conditions. Methods Cultured HPMCs were divided into normal control group, high glucose group and high glucose plus ligustrazine group （10,20,40mg/L）. Semi-quantitative reverse transcription-polymerase chain reaction was used to detect the expresssion of VEGF mRNA in HPMCs. Protein of VEGF in cultured supernatants was measured by enzyme linked immunosorbent assay（ELISA）. Cell protein concentration was measured by trace biein-choninic acid method to correct the ELISA results. Results Ligustrazine could significantly decrease the high glucose-induced VEGF expression in a dose-dependent manner both at protein and gene levels. Conclusion Ligustrazine may reduce VEGF expression in HPMCs cultured under high glucose conditions. Reduction of VEGF may prevent extracellular matrix gathering on peritoneum to slow the progress of peritoneal fibrosis associated with peritoneal dialysis.