过氧化氢对人小梁细胞内质网应激ATF4-CHOP通路的影响  

The Effect of H_2O_2 on Endoplasmic Reticulum Stress ATF4-CHOP Signaling Pathway in Human Trabecular Meshwork Cells

作　　者：陈晨[1] 朱艳[2] 李霞[1] 李珺[1] 曹智[1] 滕兴波 朱玉广[2] CHEN Chen ZHU Yan LI Xia LI Jun CAO Zhi TENG Xing-bo ZHU Yu-guang(Department of Ophthalmology, Weifang Medical University, Weifang 261053, China Ophthalmic Center, the Affiliated Hospital of Weifang Medical University)

机构地区：[1]潍坊医学院眼科学教研室,山东潍坊261053 [2]潍坊医学院附属医院眼科中心

出　　处：《潍坊医学院学报》2016年第5期379-381,共3页Acta Academiae Medicinae Weifang

基　　金：山东省卫生厅科技计划资助项目(课题编号:2015WS0047)

摘　　要：目的利用过氧化氢(H_2O_2)构建人小梁细胞体外氧化应激模型,观察H_2O_2对人小梁细胞内质网应激蛋白ATF4,CHOP及其mRNA表达的变化,探讨内质网应激(ERS)ATF4-CHOP信号通路在小梁细胞氧化损伤中的作用。方法利用600μmol/L H_2O_2构建人小梁细胞氧化应激模型。实验分为对照组、H_2O_26h组、H_2O_212h组和H_2O_224h组,各实验组小梁细胞分别以H_2O_2作用0,6,12及24h后。利用实时定量PCR检测人小梁细胞ATF4和CHOP mRNA表达,利用Western blotting法检测人小梁细胞ATF4和CHOP蛋白表达。结果与对照组比较,各H_2O_2组人小梁细胞ATF4,CHOP及其mRNA表达水平升高。随H_2O_2作用时间的延长,各H_2O_2组人小梁细胞ATF4,CHOP及其mRNA表达水平明显升高(FmRNA=17.813,13.416;F蛋白=8.856,10.082,P<0.05)。结论 H_2O_2明显促进培养的人小梁细胞ATF4,CHOP及其mRNA的表达,H_2O_2可诱导人小梁细胞发生内质网应激,内质网应激ATF4-CHOP信号通路参与了小梁细胞的氧化损伤。Objective To establish a cell oxidative stress model in vitro by using H202 in human trabecular meshwork cells （ HTCs） ,to study the effects of H202 on expressions of endoplasmic reticulum stress（ ERS） protein ATF4 and CHOP and its mRNA in HTCs and to explore the role of ERS ATF4-CHOP signaling pathway in the oxidative damage of HTCs, Methods The HTCs were cultured and pas- saged. The oxidative stress models of the 3 rd generation HTCs were established with 600 txmol/L Hl 02 treatment. The experiments were divid- ed into control group, H2 O2 6h group, H2 O2 12h group and H2 O2 24h group. The HTCs in above groups were treated with H2 02 for 0,6,12 and 24h, respectively. The expression of ATF4 and CHOP mRNA were analysed by Real-time PCR. The expression of ATF4 and CHOP were detected by Western blotting. The data were statistically analyzed. Results Compared to the control group, the expression levels of ATF4, CHOP and mRNA in the H202 groups increased. And the ATF4, CHOP and mRNA expression levels in the H202 group tended to increase gradually, along with the prolongation of the H2 O2 action time （ FmRNA = 17.813,13. 416 ; Fprotein = 8. 856,10. 082, all P 〈 0. 05 ）. Conclusion Exogenous H2 02 can significantly increase the expression of ATF4, CHOP and its mRNA in HTCs. H2 O2 can induce the endoplasmic retic- ulum stress in the human trabecular cells, and the ATF4-CHOP signaling pathway is involved in the oxidative damage of the trabecular cells. The studies suggest that ERS pathway may provide a novel target for intraocular pressure （ lOP ） regulation of HTCs.

关 键 词：过氧化氢 氧化应激 内质网应激 小梁细胞 ATF4-CHOP信号通路 

分 类 号：R775.1[医药卫生—眼科]