Ero1α在缺氧复氧致H9C2细胞内质网应激时的表达变化及意义

中国临床药理学与治疗学 ›› 2017, Vol. 22 ›› Issue (9): 972-977.

Ero1α在缺氧复氧致H9C2细胞内质网应激时的表达变化及意义

刘悦¹，刘媛媛¹，张妮¹，曹释露²，张晓京³，来丽娜³

¹长治医学院2013级教改班，²第二临床学院1301班，³药理教研室，长治 046000，山西

收稿日期:2017-03-27 修回日期:2017-07-13 出版日期:2017-09-26 发布日期:2017-09-30
通讯作者: 来丽娜，女，研究生，教授，研究方向：心血管药理学。 Tel：03553151022 E-mail：lailina@126.com
作者简介:刘悦，女，临床专业本科四年级学生。 E-mail：2927427573@qq.com
基金资助:
国家级大学生创新创业训练项目（201610117005），长治医学院创新团队项目（CX201409）

Change and significance of Ero1α expression on endoplasmic reticulum stress induced by hypoxia/reoxygenation in rat cardiomycytes

LIU Yue¹, LIU Yuanyuan ¹, ZHANG Ni ¹, CAO Shilu², ZHANG Xiaojing ³, LAI Lina³

¹Department of Reform of Education and Teaching Class of 2013 year, ² the Second Clinical College Class of 1301, ³Department of Pharmacology, Changzhi Medical College, Changzhi 046000, Shanxi, China

Received:2017-03-27 Revised:2017-07-13 Online:2017-09-26 Published:2017-09-30

摘要/Abstract

摘要：

目的: 观察内质网氧化还原酶（Endoplasmic oxidoreductin-1-like protein alpha，Ero1α）表达与内质网应激(endoplasmic reticulum stress，ERS)、细胞凋亡的关系。方法: 将培养的H9C2心肌细胞随机分组：对照组(Control组)、缺氧/复氧1、3、6、12 h组(H/R1组，H/R3组，H/R6组，H/R12组)、4-苯基丁酸钠预处理+缺氧复氧6 h组(4PBA+H/R6组)。对照组心肌细胞正常培养至实验结束，H/R组行缺氧3 h后复氧1、3、6、12 h，4PBA+H/R6组分别于缺氧前2 h给4PBA再行缺氧/复氧6 h过程。Hoechst33258染色荧光显微镜下观察细胞凋亡情况。Western blot测定Ero1α，内质网应激蛋白葡萄糖调节蛋白78(GRP78)、CCAAT/增强子结合蛋白(CHOP)、半胱氨酸门冬氨酸特异性蛋白酶12 (caspase-12)的表达。结果: 与对照组比较，Ero1α蛋白表达水平随着复氧时间的延长Ero1α表达量逐渐升高(P<0.01)，在复氧3 h明显升高，6 h达到高点。Grp78在缺氧复氧后即明显升高（P<0.01），在复氧3 h达到高峰。细胞凋亡率也显著升高，其中复氧6 h最为显著。以缺氧3 h复氧6 h作为观测点，H/R6组Ero1α、GRP78、CHOP、Caspase12蛋白表达较对照组明显增加(P<0.01)；给予4PBA干预后，Ero1α、GRP78、CHOP、caspase-12蛋白表达较H/R6组明显下降(P<0.05，P<0.01)。结论: 缺氧复氧能够诱导心肌细胞凋亡及内质网应激，缺氧复氧诱导大鼠心肌细胞内质网应激时Ero1α的表达可出现相应的变化并与细胞凋亡相关。

关键词: Ero1α, 缺氧复氧, 内质网应激, 凋亡, 心肌细胞

Abstract:

AIM: To explore the relationship between Ero1α expression and ERS and apoptosis in rat hypoxia/reoxygenation cardiomyocyte model. METHODS: The H9C2 cardiomyocytes were randomly divided into groups: control group, hypoxia /reoxygenation 1 h, 3 h, 6 h, 12 h group (H/R1 group, H/R3 group, H/R6 group, H/R12 group), 4-PBA+H/R6 group. H9C2 cardiomyocytes of control group were cultured under normal condition till the end of experiment. Cardiomyocytes of H/R group underwent hypoxia for 3 hours followed by reoxygenation for 1 h, 3 h, 6 h, 12 h. 4PBA were given 2 hours before hypoxia in the 4PBA+H/R6 group. Hoechst 33258 staining was used to observe the apoptosis of cells under fluorescence microscope. The protein expression of Ero1α, glucose-regulated proteins 78(Grp78), C/EBP homologous protein (CHOP), caspase-12 were detected by Western blot. RESULTS: Compared with the control group, the expression level of Ero1α protein increased with the time of reoxygenation. The expression of Ero1α increased significantly at 3 h after reoxygenation and reached high point at 6 h(compared with the control group, P<0.01).Grp78 increased significantly after hypoxia and reoxygenation (compared with the control group, P<0.01), reached a peak at 3 h after reoxygenation. The apoptosis rate was also significantly increased, of which the most significant was at reoxygenation 6 h. Take hypoxia 3 h reoxygenation 6 h as the observation point, the expression of Ero1α, GRP78, CHOP and Caspase-12 in H/R6 group was significantly higher than that in control group (P<0.01). After administration of 4PBA, Ero1α, GRP78, CHOP, Caspase-12 protein expression was significantly lower than that in H/R6 group (P<0.05, P<0.01). CONCLUSION: Hypoxia/reoxygenation can induce cardiomyocyte apoptosis and endoplasmic reticulum stress. The expression of Ero1α in cardiomyocytes induced by hypoxia/reoxygenation can be changed. The expression of Ero1α is related to apoptosis.

Key words: Ero1α, hypoxia/reoxygenation, endoplasmic reticulum stress, apoptosis, cardiomycytes

中图分类号:

刘悦，刘媛媛，张妮，曹释露，张晓京，来丽娜. Ero1α在缺氧复氧致H9C2细胞内质网应激时的表达变化及意义[J]. 中国临床药理学与治疗学, 2017, 22(9): 972-977.

LIU Yue, LIU Yuanyuan, ZHANG Ni, CAO Shilu, ZHANG Xiaojing, LAI Lina. Change and significance of Ero1α expression on endoplasmic reticulum stress induced by hypoxia/reoxygenation in rat cardiomycytes[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(9): 972-977.

参考文献

［1］邹麓,贾大林.内质网应激参与调节心肌缺血/再灌注损伤的新进展［J］.解剖科学进展,2016,2:207-209.
［2］Liu X, Wang M, Chen H, et al. Hypothermia protects the brain from transient global ischemia/reperfusion by attenuating endoplasmic reticulum response-induced apoptosis through CHOP［J］. PLoS One,2013, 8(1): e53431.
［3］王李瑶,路静,张建忠,等.内质网氧化还原酶1-Lα在高血压血管内皮损伤中的作用［J］.广东医学,2014,35(6):824-827.
［4］赵丽,曹成建,刘现梅,等.ERO1α及其DNA甲基化在同型半胱氨酸抑制肝细胞增殖中的作用［J］.中国药理学通报,2014,30(12) :1743-1747.
［5］赵帅,李庆玲,韩旭,等. 离体心肌细胞损伤模型研究进展［J］.长春中医药大学学报,2015,31(3):650-653.
［6］Parmar VM, Schroder M. Sensing endoplasmic reticulum stress［J］.Adv Exp Med Biol, 2012, 738: 153-168.
［7］Abas I, Ron D. Integrating the mechanisms of apoptosis induced by endoplasmic reticulum stress［J］. Nat Cell Biol, 2011,13(3):184-190.
［8］孔曼,陈娟,周洁,等. 内质网应激介导过氧化氢诱导的心肌细胞凋亡［J］. 华中科技大学学报(医学版),2012,41( 3):253-257.
［9］Decuype JP, Monaco G, Bultynck G, et al.The IP3 receptor-mitochondria connection in apoptosis and autophagy［J］.Biochim Biophys Acta,2011,1813(5): 1003-1013.
［10］汪和贵,张明超,胡雨龙,等.地高辛抗血清对缺血再灌注心肌细胞凋亡的影响及机制［J］.中国临床药理学与治疗学,2016 ,21( 5) : 486-489.
［11］Ayala P, Montenegro J, Vivar R. ttenuation of endoplasmic reticulum stress using the chemical chaperone 4-phenylbutyric acid prevents cardiac fibrosis induced by isoproterenol ［J］．Exp Mol Pathol,2012,92(1):97-104.

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