摘要
目的研究Gjb2在高糖诱导的HEI-OC1细胞株凋亡与氧化应激中的作用。方法高血糖处理HEI-OC1细胞株后,RT-qPCR检测了Gjb2的表达。Tunel实验和流式细胞术检测HEI-OC1细胞株凋亡,RT-qPCR和Western Blot检测了凋亡相关基因和蛋白的表达。采用流式细胞术检测高糖处理后HEI-OC1细胞株内ROS的生成,RT-qPCR和Western blot检测了SOD2和catalase的mRNA和蛋白表达。结果 HEI-OC1细胞株经高糖处理后,凋亡率显著升高,BCL2的mRNA表达显著降低,而BAX、Caspase-3、Caspase-9的mRNA表达则显著升高,同时BCL2的蛋白表达显著降低,而BAX、Cleaved-Caspase-3、Cleaved-Caspase-9的蛋白表达则显著升高。氧化应激检测结果显示高糖处理后细胞内ROS水平显著升高,同时伴随着SOD2和catalase的mRNA和蛋白表达显著升高。高糖处理显著抑制了Gjb2的表达,而在HEI-OC1细胞株中过表达Gjb2则能抑制高糖诱导的细胞凋亡和氧化应激。结论过表达Gjb2能抑制高糖诱导的HEI-OC1细胞株凋亡和氧化应激。
Objective To study the role of Gjb2 in cell apoptosis and oxidative stress induced by high glucose using HEI-OC1 cells. Methods Expression of Gjb2 was detected by RT-qPCR after treating HEI-OC1 cells with high-glucose. HEI-OC1 cells apoptosis was detected by TUNEL assay and flow cytometry, and expression of apoptosis-related genes and proteins was detected using RT-qPCR and Western Blot. Generation of ROS in HEI-OC1 cells was detected by flow cytometry, and mRNA and protein expression of SOD2 and catalase was detected by RT-qPCR and Western blot.Result After high-glucose treatment, the rate of apoptosis in HEI-OC1 cells was significantly increased. RT-qPCR revealed remarkably decreased mRNA expression of BCL2, while mRNA expression of BAX, Caspase-3 and Caspase-9 was dramatically increased. Western Blot revealed prominently decreased level of BCL2, while that of BAX, CleavedCaspase-3 and Cleaved-Caspase-9 increased notably. In addition, ROS level in HEI-OC1 cells increased markedly after high-glucose treatment, accompanied by a significant increase in mRNA and protein expression of SOD2 and catalase.Mechanically, Gjb2 expression was inhibited by high-glucose, while overexpression of Gjb2 in HEI-OC1 cells inhibited apoptosis and oxidative stress induced by high glucose. Conclusion Overexpression of Gjb2 in HEI-OC1 cells inhibited apoptosis and oxidative stress induced by high glucose.
作者
王大华
何茜
赵凯
程思
何清
WANG Dahua;HE Qian;ZHAO Kai;CHENG Si;HE Qing(China University of Mining and Technology;Affiliated Hospital of China University of Mining and Technology;Huangshi Central Hospital,Eastern Hubei Medical Group;Huangshi Maternal and Child Health Care Hospital,Eastern Hubei Medical Group)
出处
《中华耳科学杂志》
CSCD
北大核心
2021年第6期930-937,共8页
Chinese Journal of Otology
基金
中央高校基本科研业务费专项资金资助(2020QN88)。
