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目的探讨水飞蓟宾对高糖诱导的体外培养人足细胞中血管内皮生长因子(VEGF)表达的影响。方法体外培养的人足细胞分为7组:正常糖(NG,葡萄糖5.5mmol/L)组、甘露醇对照(MN,甘露醇24.5mmol/L+葡萄糖5.5mmol/L)组、高糖(HG,葡萄糖30mmol/L)组、HG+低浓度水飞蓟宾(HG+LS,水飞蓟宾5μg/ml)组、HG+中浓度水飞蓟宾(HG+MS,水飞蓟宾10μg/ml)组、HG+高浓度水飞蓟宾(HG+HS,水飞蓟宾20μg/ml)组、HG+二甲亚砜(HG+D,DMSO1mg/ml)组。各作用48h后,RT-PCR半定量法及Western印迹法检测足细胞VEGFmRNA及蛋白表达水平。结果体外培养人足细胞表达基础水平的VEGF,HG刺激48h后足细胞VEGF基因和蛋白表达均显著增加(P<0.05);与HG组比较,加用5～20μg/ml水飞蓟宾均可抑制HG所诱导的VEGF基因和蛋白表达(P<0.05),并呈剂量依赖性。结论 HG可使足细胞表达VEGF增加,水飞蓟宾可通过抑制HG诱导足细胞产生过多VEGF而具有潜在保护足细胞和防治糖尿病肾病的作用。
Objective To investigate the effect of silybin on the expression of vascular endothelial growth factor (VEGF) in cultured human podocytes induced by high glucose. Methods Human podocyte cultured in vitro were divided into seven groups: normal glucose (NG, glucose 5.5mmol / L), mannitol control (MN, mannitol 24.5mmol / L + glucose 5.5mmol / (HG + LS, silybin 5μg / ml), HG + medium concentration silybin (HG + MS, silybin 10μg / ml) , HG + high concentration silybin (HG + HS, silybin 20μg / ml) group, HG + dimethyl sulfoxide (HG + D, DMSO1mg / ml) group. After 48 hours, the mRNA and protein expression of VEGF in podocytes were detected by semi-quantitative RT-PCR and Western blotting. Results The expression of VEGF gene and protein in podocytes was significantly increased after VEGF and HG stimulation for 48 h in vitro (P <0.05). Compared with HG group, the addition of 5 ~ 20 μg / ml silybin Inhibited the HG-induced VEGF gene and protein expression (P <0.05), and in a dose-dependent manner. Conclusions HG can increase the expression of VEGF in podocytes. Silybin can potentially protect podocytes and prevent diabetic nephropathy by inhibiting HG-induced excessive VEGF production in podocytes.