AIM:To observe the expression of hypoxia-inducible factor-1α (HIF-1α) , vascular endothelial growth factor A (VEGFA) and vascular endothelial cadherin (VE-cadherin) in cultured rat renal glomerular endothelial cells (rRGECs) exposed to glucose at different concentrations in vitro, and to verify the hypothesis that salidroside attenuates high glucose (HG) -induced injury of rRGECs by boosting HIF-1αlevel.METHODS:rRGECs were divided into 4group:normal glucose (NG) group, HG groups, hypertonic group and salidroside+HG group.The viability of rRGECs was measured by MTT assay.The mRNA expression of VEGFA, VE-cadherin and HIF-1αwas detected by RT-qPCR.The protein expression of HIF-1αwas determined by Western blot.RESULTS:Compared with NG group, the mRNA and protein expression of HIF-1αwas increased when the rRGECs were treated with glucose at concentration of 20 mmol/L for 24h (P<0.01).Compared with NG group, the mRNA expression of HIF-1αwas decreased in HG groups for 120 h (P<0.05).Compared with NG group, the mRNA expression of VE-cadherin was significantly down-regulated in HG groups for24 h or 120 h (P<0.05).Compared with NG group, the mRNA expression of VEGFA was increased in HG groups at 24h (P<0.05) , while the mRNA expression of VEGFA was decreased at 120 h (P<0.05).Compared with NG group, no statistical difference in the mRNA expression levels of HIF-1α, VE-cadherin and VEGFA in DM group was observed.Compared with HG group, salidroside promoted the viability of rRGECs (P<0.01) , and up-regulated the mRNA expression of HIF-1αand VE-cadherin, and the protein expression of HIF-1α (P<0.05).CONCLUSION:High glucose regulates HIF-1αexpression in rRGECs in connection with cell viability, the concentration of glucose, the culture time and HIF/VEGF signaling.Salidroside promotes rRGEC growth against high glucose-induced cell apoptosis via up-regulating the expression of HIF-1α.%目的:观察体外不同浓度葡萄糖对大鼠肾小球内皮细胞 (rRGECs) 表达低氧诱导因子1α (HIF-1α) 、血管内皮生长因子A (VEGFA) 及血管内皮钙黏素 (VE-cadherin) 的影响, 探讨红景天苷减轻高糖诱导rRGECs损伤的作用及可能相关机制.方法:体外培养rRGECs, 分为正常糖组、高糖 (20、30和50 mmol/L) 组、高渗组及红景天苷+高糖组.采用MTT法检测rRGECs的活力;RT-qPCR法检测rRGECs内HIF-1α、VEGFA及VE-cadherin的mRNA表达;Western blot法检测rRGECs内HIF-1α蛋白的表达.结果:与正常糖组相比, 培养24 h后, 高糖 (20mmol/L) 组rRGECs HIF-1α的mRNA及蛋白表达均上调 (P <0.05) ;培养120 h后, 高糖组HIF-1αmRNA表达下调 (P <0.05) .与正常糖组相比, 培养24 h和120 h后, 高糖组rRGECs内VE-cadherin的mRNA表达下调 (P <0.05) .与正常糖组相比, 培养24 h后, 高糖组rRGECs内VEGFA的mRNA表达上调 (P <0.05) ;培养120 h后, 高糖组rRGECs内VEGFA的mRNA表达下调 (P <0.05) .与正常糖组相比, 培养24 h和120 h后, 高渗组rRGECs内HIF-1α、VE-cadherin及VEGFA的mRNA表达无变化.与高糖组相比, 培养24 h后, 红景天苷 (50μmol/L) 组rRGECs的活力增加 (P <0.01) , 且细胞内HIF-1α和VE-cadherin的mRNA及HIF-1α蛋白表达均上调 (P <0.05) .结论:体外高糖培养能影响rRGECs表达HIF-1α, 可能与细胞活力、葡萄糖的浓度、作用时间及HIF/VEGF通路有关.红景天苷能减轻高糖诱导的rRGECs损伤, 其机制可能与增加rRGECs内HIF-1α的表达有关.
展开▼
