干扰或过表达PDIA3对过氧化氢诱导的星形胶质细胞损伤的影响  

Effects of interference or overexpression of PDIA3 on hydrogen peroxide-induced astrocyte damage

作　　者：雷雨广 徐继鹏[3] 陈超[1,2] 张伟丽 LEI Yu-Guang;XU Ji-Peng;CHEN Chao(Department of Pathology,the Second People's Hospital of Xinyang City,Xinyang 464000,Henan,China)

机构地区：[1]信阳市第二人民医院病理科,河南信阳464000 [2]信阳职业技术学院护理学院 [3]信阳职业技术学院附属医院皮肤科

出　　处：《中国老年学杂志》2020年第3期585-589,共5页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(No.81602079)

摘　　要：目的探讨干扰或过表达蛋白质二硫键异构酶(PDI)A3对过氧化氢诱导的星形胶质细胞损伤的影响及其作用机制。方法体外培养星形胶质细胞,构建PDIA3干扰及过表达载体稳定株系,采用过氧化氢(H 2O 2)诱导建立星形胶质细胞损伤模型。实验将星形胶质细胞分为对照组(NC组)、H 2O 2组、H 2O 2+si-con组、H 2O 2+si-PDIA3组、H 2O 2+pcDNA组、H 2O 2+pcDNA-PDIA3组。实时荧光定量-聚合酶链反应(qRT-PCR)检测PDIA3 mRNA表达;Western印迹检测PDIA3及p38丝裂原活化蛋白激酶(p38MAPK)信号通路蛋白表达;噻唑蓝(MTT)法检测细胞活力;流式细胞术检测细胞凋亡。收集各组星形胶质培养基上清液检测细胞乳酸脱氢酶(LDH)释放率及肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6水平。结果与NC组相比,H 2O 2组星形胶质细胞中PDIA3 mRNA及蛋白表达水平均显著升高(P<0.05);H 2O 2组细胞LDH释放量、TNF-α、IL-6水平、细胞凋亡率及p-p38蛋白表达均显著升高(P<0.05),干扰PDIA3表达后明显升高(P<0.05),PDIA3过表达后明显降低(P<0.05);H 2O 2组细胞存活率显著降低(P<0.05),干扰PDIA3表达后明显降低(P<0.05),PDIA3过表达后明显升高(P<0.05)。结论PDIA3过表达可改善H 2O 2诱导的星形胶质细胞氧化应激及炎症损伤,其可能通过抑制p38MAPK信号通路激活而发挥作用。Objective To investigate the effect of interfering or overexpressing protein disulfide isomerase(PDI)A3 on hydrogen peroxide-induced astrocyte damage and its mechanism.Methods Astrocytes were cultured in vitro to construct a PDIA3 interference and over-expression vector stable strain,and astrocytic cell injury model was established by hydrogen peroxide induction.The astrocytes were divided into NC group,H 2O 2 group,H 2O 2+si-con group,H 2O 2+si-PDIA3 group,H 2O 2+pcDNA group and H 2O 2+pcDNA-PDIA3 group.qRT-PCR was used to detect the expression of PDIA3 mRNA,Western blot was used to detect the expressions of PDIA3 and p38MAPK signaling pathway protein.MTT method was used to detect the viability of each group of cells.Apoptosis was detected by flow cytometry.The supernatants of each group of astrocytes were collected and tested for the release rate of LDH and the levels of TNF-αand IL-6.Results Compared with the NC group,the expression of PDIA3 mRNA and protein in the astrocytes of H 2O 2 group was significantly increased(P<0.05).The LDH release,TNF-α,IL-6 level,apoptosis rate and p-p38 protein expression in H 2O 2 group were significantly increased(P<0.05),and the expression of PDIA3 was significantly increased(P<0.05).PDIA3 significantly decreased after overexpression(P<0.05).The survival rate of H 2O 2 cells was significantly decreased(P<0.05),and the expression of PDIA3 was significantly decreased(P<0.05),and PDIA3 was significantly increased after overexpression(P<0.05).Conclusions Overexpression of PDIA3 could improve hydrogen peroxide-induced astrocyte oxidative stress and inflammatory damage,which might play a role in inhibiting the activation of the p38 MAPK signaling pathway.

关 键 词：蛋白质二硫键异构酶A3 星形胶质细胞 p38丝裂原活化蛋白激酶(MAPK)信号通路 氧化损伤 

分 类 号：R741[医药卫生—神经病学与精神病学]