miRNA378*对柯萨奇B3病毒感染心肌细胞凋亡、网腔钙结合蛋白、内质网应激及信号通路因子的作用

杨焱; 王羽; 崔晓雪; 邱祥春; 魏成喜; 赵明; 刘永峰

doi:10.13201/j.issn.1001-1439.2016.12.020

miRNA378*对柯萨奇B3病毒感染心肌细胞凋亡、网腔钙结合蛋白、内质网应激及信号通路因子的作用

- 1 内蒙古民族大学附属医院(内蒙古通辽, 028000);
- 2 内蒙古民族大学临床医学院;
- 3 内蒙古民族大学医学院药物化学与药理学研究所
基金项目:
内蒙古自治区自然科学基金资助课题(No:2015MS08153)

详细信息

通讯作者: 刘永峰,E-mail:dr.liudanial@gmail.com

中图分类号: R331.3

收稿日期: 2016-05-30

修回日期: 2016-08-05

The effect of miRNA378* on the myocardial cells apoptosis,calumenin,endoplasmic reticulum stress and signaling pathway factor caused by Coxsackie virus

- 1 The Inner-mongolia National University Affiliated Hospital, Tongliao, Inner-mongolia, 028000, China;
- 2 The Inner-mongolia National University Medical College;
- 3 The Inner-mongolia National University medicinal chemistry and Pharmacology Institute

More Information

Corresponding author: LIU Yongfeng, E-mail: dr.liudanial@gmail.com

Received Date: 30 May 2016

Revised Date: 05 August 2016

摘要

摘要: 目的:探讨上调miRNA378*表达对柯萨奇B3病毒(CVB3)感染心肌细胞凋亡、网腔钙结合蛋白、内质网应激及信号通路因子的作用。方法:实验分4组:对照组(正常细胞)、CVB3感染组(正常细胞+ CVB3)、miRNA378*过表达对照组(正常细胞+CVB3+转染miRNA378*空表达质粒)、miRNA378*过表达组(正常细胞+ CVB3+转染miRNA378*过表达质粒)。原代培养乳鼠心肌细胞,采用免疫组织化学方法检测培养乳鼠心室肌细胞α-SMA蛋白,慢病毒质粒转染心室肌细胞,除对照组外,其他各组心肌细胞感染CVB3,采用TUNEL技术检测各组心肌细胞凋亡率;用Western blotting技术检测各组心肌细胞网腔钙结合蛋白、内质网应激伴侣蛋白GRP78及内质网应激信号通路因子PERK、P-PERK 、eIF2α、ATF4、CHOP表达。结果:与CVB3感染组比较,miRNA378*过表达组心肌细胞凋亡率明显减少,网腔钙结合蛋白表达增加,而GRP78、P-PERK 、eIF2α、ATF4、CHOP表达均减少(均P<0.01),PERK表达差异无统计学意义。结论:上调CVB3感染心肌细胞miRNA378*表达可引起心肌细胞凋亡减少,网腔钙结合蛋白表达增多,进而缓解内质网应激,并抑制内质网应激凋亡信号通路因子表达。
- miRNA378* /
- 柯萨奇B3病毒 /
- 内质网应激 /
- 网腔钙结合蛋白 /
- 凋亡
Abstract: Objective: To investigate the effect of miRNA378* on the myocardial cells apoptosis, calumenin protein, endoplasmic reticulum stress and signaling pathway caused by Coxsackie virus. Method: The suckling mouse myocardium of primary culture were randomly divided into control group, Coxsackie virus B3 infecton group, lentivirus infection miRNA378* overexpression control group, lentivirus infection miRNA378* over expression group. Neonatal rat cardiomyocytes were cultured, α-SMA was monitored by immunohistochemical method, ventricular myocytes were transfered by lentivirus plasmids. Excepted control group, the myocardial cells of other groups infected with Coxsackie virus B3. TUNEL was used to detect the apoptosis rate of myocardial cells and the Western blotting was used to detect the expression change of calumenin,endoplasmic reticulum stress chaperone protein GRP78 and endoplasmic reticulum stress signaling pathway factors PERK, P-PERK, eIF2 alpha, ATF4, CHOP. Result: Compared with Coxsackie virus infection group, myocardial cell apoptosis rate was significantly decreased, calumenin protein expression was increased, and GRP78, P-PERK, eIF2 alpha, ATF4, CHOP expression were decreased in miRNA378* overexpression group (all P<0.01). While PERK expression had no statistical significance between the two groups. Conclusion: Upregulation of Coxsackie virus B3 infected myocardial cells miRNA378* expression could decrease myocardial cell apoptosis, and increase calumenin expression, thereby alleviate the endoplasmic reticulum stress, and inhibit the expression of endoplasmic reticulum stress mediated apoptosis signaling pathway.
- miRNA378* /
- Coxsackie virus B3 /
- endoplasmic reticulum stress /
- calumenin /
- apoptosis

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