血竭素高氯酸盐抑制高糖诱导的人肾小球系膜细胞中SGK1和FN的表达

Dracorhodin perchlorate inhibit high glucose induce serum and glucocorticoid induced protein kinase 1 and fibronectin expression in human mesangial cells

目的:观察血竭素高氯酸盐(dracorhodin perchlorate,DP)对高糖环境下培养的人肾小球系膜细胞(human me-sangial cells,HMC)中血清和糖皮质激素诱导的蛋白激酶1(serum and glucocorticoid induced protein kinase1,SGK1)和纤连蛋白(fibronectin,FN)的抑制作用,探讨其防治糖尿病肾病肾脏纤维化的作用与机制。方法:将体外培养的HMC分为正常糖组(NG,5.5mmol.L-1D-葡萄糖)、正常糖+DP低剂量组(NG+LDP,5.5mmol.L-1D-葡萄糖+7.5μmol.L-1DP)、正常糖+DP高剂量组(NG+HDP,5.5mmol.L-1D-葡萄糖+15μmol.L-1DP)、高糖组(HG,25mmol.L-1D-葡萄糖)、高糖+DP低剂量组(HG+LDP,25mmol.L-1D-葡萄糖+7.5μmol.L-1DP)、高糖+DP高剂量组(HG+HDP,25mmol.L-1D-葡萄糖+15μmol.L-1DP),作用24h后,采用real-time PCR法检测各组细胞中SGK1,FN mRNA的水平,并采用间接免疫荧光和Westernblot方法检测细胞中SGK1和FN蛋白的表达。结果:在正常糖组HMC中有基础量的SGK1,FN mRNA及蛋白的表达,经7.5μmol.L-1的DP作用24h后,SGK1和FN mRNA水平及蛋白与NG组相比无明显差异,但经15μmol.L-1的DP作用24h后,SGK1和FN mRNA水平及蛋白较NG组明显下降;与NG组相比较,HG组中HMC中的SGK1,FN mRNA及蛋白水平显著性增加(P<0.01);与HG组相比较,HG+LDP组和HG+HDP组中,HMC的SGK1,FN的mRNA及蛋白水平显著性下降(P<0.01)。结论:血竭素高氯酸盐抑制高糖诱导的人肾小球系膜细胞中SGK1和FN的表达,这可能是其防治DN肾纤维化作用的部分机制。

Objective：To investigate the effect of dracorhodin perchlorate（DP）on inhibiting high glucose-induced serum and glucocorticoid induced protein kinase 1（SGK1）and fibronectin（FN）expression in human mesangial cells（HMC）,and its mechanism of prevention and treatment on renal fibrosis in diabetic nephropathy（DN）.Method：The HMC were divided into normal glucose group（NG group,5.5 mmol·L^-1D-glucose）,normal glucose＋low DP group（NG＋LDP group,5.5 mmol·L^-1 D-glucose＋7.5 μmol·L^-1 DP）,normal glucose＋high DP group（NG＋HDP group,5.5 mmol·L^-1 D-glucose＋15 μmol·L^-1 DP）,high glucose group（HG group,25 mmol·L^-1D-glucose）,high glucose＋low DP group（HG＋LDP group,25 mmol·L^-1 D-glucose＋ 7.5 μmol·L^-1 DP）and high glucose＋high DP group（HG＋HDP group,25 mmol·L^-1 D-glucose＋ 15 μmol·L^-1 DP）.Each group was examined at 24 hours.The levels of SGK1 and FN mRNA was detected by real-time fluorescence quantitative PCR,and the expression of SGK1 and FN protein was detected by Western blot or indirect immunofluorescence.Result：A basal level of SGK1 and FN in HMC were detected in NG group,and the level of SGK1 and FN mRNA and protein were not evidently different compared to that of NG group adding 7.5 μmol·L^-1 DP for 24 hours.On the other hand,the levels of SGK1 and FN mRNA and protein were obviously decreased by adding 15 μmol·L^-1 DP for 24 hours.Compared to NG group,the levels of SGK1 and FN mRNA and protein were increased in HG group after stimulating for 24 hours（P〈0.01）.Compared to HG group,the level of SGK1 and FN mRNA and protein were evidently reduced in HG＋LDP and HG＋HDP groups by adding 7.5 μmol·L^-1 DP and 15 μmol·L^-1 DP for 24 hours（P〈0.01）.Conclusion：Dracorhodin perchlorate can inhibit high glucose-induced serum and glucocorticoid induced protein kinase 1（SGK1）and fibronectin（FN）expression in human mesangial cells,and this may be part of the mechanism of preventing and treating renal fibrosis of DN.