石斛酚对高糖诱导的人视网膜微血管内皮细胞的保护作用

Protective effect of gigantolon human retinal microvascular endothelial cells induced by high glucose

目的:探讨石斛酚对高糖诱导的人视网膜微血管内皮细胞(HRMECs)的醛糖还原酶(AR)通路及氧化应激的保护作用。方法:将体外培养的HRMECs分为5组:正常组(葡萄糖5. 5mmol/L)、甘露醇组(葡萄糖5. 5mmol/L+甘露醇24. 5mmol/L)、高糖组(葡萄糖30mmol/L)、石斛酚组(葡萄糖30mmol/L+石斛酚5μmol/L)、依帕司他组(葡萄糖30mmol/L+依帕司他1μmol/L)。通过MTS实验检测细胞活力,DCFH-DA荧光探针法检测细胞内活性氧(ROS)水平; Western blotting法检测核因子κB(NF-κB)的蛋白表达; qRT-PCR法检测AR、HIF-1α及VEGF的mRNA表达。结果:DCFH-DA荧光探针法结果显示石斛酚可明显降低高糖环境下细胞内的ROS水平。与高糖组相比,石斛酚组NF-κB的蛋白表达,AR、HIF-1α及VEGF的mRNA表达也都明显下降。依帕司他组的AR及NF-κB表达趋势与石斛酚组一致。结论:石斛酚与依帕司他都可抑制AR/NF-κB通路的激活,同时石斛酚还可抑制高糖诱导的HRMECs氧化应激反应。石斛酚在糖尿病视网膜病变的发生发展中可能有一定的保护作用,可能帮助减少新生血管的生成。

AIM:To investigate the protective effects of gigantol on aldose reductase(AR)pathway and oxidative stress induced by high glucose in human retinal microvascular endothelial cells(HRMECs).METHODS:HRMECs were cultured and divided into the five groups:normal glucose group,mannitol group,high glucose group,gigantol group and epalrestat group.Cell vitality was detected by the MTS assay,reactive oxygen species(ROS)levels were detected by DCFH-DA fluorescent probe,protein expressions of nuclear factor kappa B(NF-κB)were observed by Western blotting,mRNA levels of AR,hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF)were measured by qRT-PCR.RESULTS:DCFH-DA fluorescent probe levels showed that gigantol reduced ROS generation obviously,and protein expressions of NF-κB,mRNA expressions of AR,HIF-1α and VEGF were all decreased.The expression trends of AR and NF-κB in epalrestat group were consistent with gigantol group.CONCLUSION:Gigantol and epalrestat could inhibit the activation of AR/NF-κB pathway,and gigantol also could inhibit high glucose induced oxidative stress and in HRMECs,which may have a protective effect on diabetic retinopathy(DR),and improve angiogenesis.