人ether-α-go-go钾通道减少肝脏的内质网应激和凋亡

doi:10.3969/j.issn.1006-7795.2019.01.009

首都医科大学学报 ›› 2019, Vol. 40 ›› Issue (1): 45-52.doi: 10.3969/j.issn.1006-7795.2019.01.009

• 糖尿病研究从基础到临床 • 上一篇下一篇

人ether-α-go-go钾通道减少肝脏的内质网应激和凋亡

卢晶, 沈涵, 程呈, 刘敬怡, 朱晓蓉, 谢荣荣, 袁明霞, 杨金奎

首都医科大学附属北京同仁医院内分泌科糖尿病防治研究北京市重点实验室北京市糖尿病研究所, 北京 100730

收稿日期:2018-11-08 出版日期:2019-01-21 发布日期:2019-01-23
通讯作者: 杨金奎 E-mail:jinkui.yang@gmail.com
基金资助:
国家自然科学基金（81800688，81471014，81561128015），北京市属医院科研培育计划（PX2019006），首都医科大学附属北京同仁医院科研基金（TRYY-KYJJ-2016-013）。

Effect of human ether-α-go-go related gene channels on hepatic endoplasmic reticulum stress and apoptosis

Lu Jing, Shen Han, Cheng Cheng, Liu Jingyi, Zhu Xiaorong, Xie Rongrong, Yuan Mingxia, Yang Jinkui

Department of Endocrinology, Beijing Tongren Hospital, Capital Medical University;Beijing Key Laboratory of Diabetes Research and Care;Beijing Diabetes Institute, Beijing 100730, China

Received:2018-11-08 Online:2019-01-21 Published:2019-01-23
Supported by:
This study was supported by National Natural Science Foundation of China (81800688,81471014,81561128015), Beijing Municipal Administration of Hospitals Incubating Program (PX2019006), Foundation of Beijing Tongren Hospital, Capital Medical University (TRYY-KYJJ-2016-013).

摘要/Abstract

摘要： 目的研究人ether-α-go-go（human ether-α-go-go related gene，hERG）钾通道在肝细胞中与内质网应激和凋亡的关系。方法选取雄性20周龄hERG基因敲除（knockout，KO）及同窝野生（wild type，WT）小鼠，测量体质量、摄食量；肝脏石蜡切片HE染色；Western blotting法检测糖异生关键酶葡萄糖-6-磷酸酶（glucose-6-phosphatase，G6pase）和磷酸烯醇式丙酮酸羧激酶（phosphoenolpyruvate carboxykinase，PEPCK）在肝脏的表达水平；Western blotting和荧光定量PCR方法检测肝脏凋亡相关基因活化的半胱天冬酶半胱天冬酶3（cleaved-cysteine aspartyl protease 3，cleaved-caspase 3），B细胞淋巴瘤因子2（B-cell lymphoma 2，Bcl2）和Bcl2相关X蛋白（Bcl2 associated X protein，Bax）表达水平；同时检测内质网应激相关基因磷酸化真核细胞翻译起始因子2（phosphorylated-eukaryotic translation initiation factor 2α，p-eIF2α）、激活转录因子4（activating transcription factor 4，ATF4）以及C/EBP同源蛋白（C/EBP-homologous protein，CHOP）等基因的表达。经尾静脉给KO小鼠注射hERG慢病毒（lentivirus，LV）后，Western blotting法检测胰岛及肝脏内质网应激和凋亡相关指标。结果与WT小鼠相比，KO小鼠体质量及摄食量差异无统计学意义，肝细胞轻度肿胀，肝脏糖代谢G6Pase表达水平无明显变化，而PEPCK表达上调提示hERG敲除引起肝脏糖代谢异常。KO小鼠肝脏的内质网应激及凋亡相关指标均上调，提示KO小鼠体内存在内质网应激和凋亡。KO小鼠体内过表达hERG后，改善了小鼠的肝脏内质网应激及凋亡。结论 hERG钾通道可以通过减少肝脏内质网应激和凋亡改善肝细胞内糖代谢。

关键词: 人ether-α-go-go相关基因, 内质网应激, 凋亡, 肝脏

Abstract: Objective To study the relationship between human ether-α-go-go related gene (hERG) potassium channel and endoplasmic reticulum stress and apoptosis in hepatocytes. Methods The body weight and food intake of male 20-week-old hERG knockout (KO) and wild type (WT) mice, and HE staining of liver paraffin sections was performed. Western blotting was used to detect key enzymes of gluconeogenesis including glucose-6-phosphatase (G6pase) and phosphoenolpyruvate carboxykinase (PEPCK). Western blotting and real-time qPCR methods were used to detect liver apoptosis related genes including cleaved-cysteine aspartyl protease 3 (cleaved-caspase 3), B-cell lymphoma 2 (Bcl2) and Bcl2 associated X protein (Bax). Liver endoplasmic reticulum stress related genes including, phosphorylated-eukaryotic translation initiation factor 2α(p-eIF2α), activating transcription factor 4 (ATF4) and C/EBP-homologous protein (CHOP) were also detected. hERG lentivirus was injected via tail vein of KO mice, and Western blotting was used to detect the protein levels of islet and liver endoplasmic reticulum stress and apoptosis related indicators. Results Compared with WT mice, no statistically significant differences was observed in body weight and food intake in KO mice. G6Pase expression level has no significant change, while the up-regulation of PEPCK expression suggested that hERG knockout caused abnormal liver glucose metabolism. The endoplasmic reticulum stress and apoptosis related genes were up-regulated suggesting the presence of endoplasmic reticulum stress and apoptosis in KO mice. After the overexpression of hERG in KO mice, the liver endoplasmic reticulum stress and apoptosis were improved. Conclusion hERG potassium channel can improve glucose metabolism in liver cells with reducing liver endoplasmic reticulum stress and apoptosis.

Key words: human ether-α-go-go related gene, endoplasmic reticulum stress, apoptosis, liver

中图分类号:

卢晶, 沈涵, 程呈, 刘敬怡, 朱晓蓉, 谢荣荣, 袁明霞, 杨金奎. 人ether-α-go-go钾通道减少肝脏的内质网应激和凋亡[J]. 首都医科大学学报, 2019, 40(1): 45-52.

Lu Jing, Shen Han, Cheng Cheng, Liu Jingyi, Zhu Xiaorong, Xie Rongrong, Yuan Mingxia, Yang Jinkui. Effect of human ether-α-go-go related gene channels on hepatic endoplasmic reticulum stress and apoptosis[J]. Journal of Capital Medical University, 2019, 40(1): 45-52.

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人ether-α-go-go钾通道减少肝脏的内质网应激和凋亡

Effect of human ether-α-go-go related gene channels on hepatic endoplasmic reticulum stress and apoptosis

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