摘要
目的:探讨槐杞黄(HQH)是否通过激活Nrf2/HO-1信号通路减轻高糖诱导的HK-2细胞氧化应激.方法:将HK-2细胞随机分为:正常组、渗透压对照组、高糖组、槐杞黄组和卡托普利组.采用CCK-8法检测细胞活力;荧光探针检测ROS水平;试剂盒检测SOD活性、MDA和GSH含量;免疫荧光法检测Nrf2的表达;qRT-PCR法检测IL-6、IL-1β和TNF-α以及Nrf2/HO-1 mRNA表达水平;Western blot法检测Nrf2/HO-1通路的表达.结果:与正常组比较,高糖组细胞存活率明显降低(P<0.000 1);细胞中ROS、MDA含量升高(P<0.000 1),SOD和GSH活性降低(P<0.000 1);Nrf2的表达降低;IL-6、IL-1β和TNF-α mRNA表达量均明显升高(P<0.001);Nrf2/HO-1蛋白和mRNA表达量明显减少(P<0.001);与高糖组相比,槐杞黄组细胞存活率明显升高(P<0.05);细胞中ROS、MDA含量明显降低(P<0.000 1),SOD和GSH活性升高(P<0.01);Nrf2的表达增加;IL-6、IL-1β和TNF-α mRNA表达量明显降低(P<0.01);Nrf2/HO-1蛋白和mRNA表达量均明显升高(P<0.01).结论:槐杞黄可能通过激活Nrf2/HO-1信号通路,提高机体抗氧化水平,改善高糖诱导的HK-2细胞氧化损伤.
Abstract
Objective:This study aims to investigate the antioxidant stress effect of HK-2 cells by Huaiqihuang(HQH)through highglucose-induced HK-2 and its Nrf2/HO-1 pathway.Methods:The cells were randomly divided into five groups:nor-mal group,osmotic control group,high-glucose group,HQH group,and captopril group.The cell viability was detected by CCK-8;the level of ROS was detected by fluorescence probe;the activities of SOD,MDA,and GSH content were tested by as-say kits;the expression of Nrf2 was detected by immunofluorescence staining;the mRNA expression levels of IL-6,IL-1β,TNF-α,and Nrf2/HO-1 were detected by qRT-PCR;the expression of Nrf2/HO-1 pathway was detected by Western blotting.Results:Compared with the normal group,the cell viability of the high-glucose group was significantly decreased(P<0.000 1);the levels of ROS and MDA in the cells were increased(P<0.000 1),and the activities of SOD and GSH were decreased(P<0.000 1);The expression of Nrf2 was decreased.the mRNA expression levels of IL-6,IL-1β,and TNF-α were significantly re-duced(P<0.001);the protein and mRNA expression levels of Nrf2/HO-1 were significantly decreased(P<0.001).Compared with the high-glucose group,the cell viability of the HQH groups were significantly increased(P<0.05);the levels of ROS and MDA in the cells were significantly decreased(P<0.000 1),and the activities of SOD and GSH were increased(P<0.01);The expression of Nrf2 was increased.the mRNA expression levels of TNF-α,IL-6,and IL-1β were significantly decreased(P<0.01);the protein and mRNA expression levels of Nrf2/HO-1 were significantly increased(P<0.01).Conclusion:HQH may al-leviate highglucose-induced oxidative stress in HK-2 cells by activating the Nrf2/HO-1 signaling pathway,thus enhancing the body's antioxidant capacity and protecting the cells from oxidative damage.
维普
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