过表达c-SKI抑制异丙肾上腺素诱导的小鼠心肌纤维化

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (3): 364-369.

过表达c-SKI抑制异丙肾上腺素诱导的小鼠心肌纤维化

张颖, 向燕, 张岳, 王娟^*

新疆医科大学第五附属医院心血管内科, 新疆乌鲁木齐 830011

收稿日期:2020-04-06 修回日期:2020-06-30 出版日期:2021-03-05 发布日期:2021-03-01
通讯作者: ^*15160991696@163.com
基金资助:
新疆维吾尔自治区自然科学基金(2017D01C273)

Over-expression of c-SKI inhibits isoprenaline-induced myocardial fibrosis in mice

ZHANG Ying, XIANG Yan, ZHANG Yue, WANG Juan^*

Department of Cardiology, the Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, China

Received:2020-04-06 Revised:2020-06-30 Online:2021-03-05 Published:2021-03-01
Contact: ^*15160991696@163.com

摘要/Abstract

摘要： 目的探讨原癌基因c-SKI在异丙肾上腺素(ISO)诱导的小鼠心肌纤维化中的作用及机制。方法将小鼠分为对照组(control组)、模型组(model组)、c-SKI基因感染模型组(LV-SKI组)及对照病毒感染模型组(LV-NC组),每组10只。模型组采用皮下注射ISO 30 d,首次剂量为5 mg/kg,后续以2.5 mg/kg。2周后LV-SKI组和LV-NC组于18、21、24、27和30 d分别尾静脉注射c-SKI慢病毒和对照病毒,剂量均为100 μL,滴度1×10⁸ TU。HE染色和Masson染色观察心肌组织病理学;ELISA检测心肌组织中Ⅰ型和Ⅲ型胶原蛋白含量;Western blot检测c-SKI、间质细胞标志物波形蛋白(vimentin)、α-SMA、内皮细胞标志物CD31以及转录因子Snail、Twist及Slug的表达。结果与对照组相比,模型组小鼠心肌纤维组织中c-SKI的表达呈时间依赖性下降(P＜0.01);模型组心肌细胞排列紊乱,间质显著增多,Ⅰ型和Ⅲ型胶原蛋白含量增加(P＜0.01);心肌组织vimentin、α-SMA、Snail、Twist及Slug表达均上调,CD31蛋白表达下调(P＜0.05)。过表达c-SKI后,与LV-NC组比较,小鼠心肌结构趋于整齐,间质胶原纤维减少,心肌组织中Ⅰ型和Ⅲ型胶原蛋白减少(P＜0.01);Vimentin、α-SMA蛋白的表达下调(P＜0.05);CD31蛋白的表达上调(P＜0.05);同时转录因子Snail、Twist和Slug蛋白的表达也随之下调(P＜0.01)。结论小鼠心肌纤维化进程中c-SKI表达下调,而过表达c-SKI能够通过抑制内皮-间充质转化(End-MT)改善ISO诱导的心肌纤维化。

关键词: c-SKI, 心肌纤维化, 内皮-间充质转化(End-MT), 转录因子

Abstract: Objective To investigate the effect of c-SKI on isoprenaline(ISO)-induced cardiac fibrosis in mice. Methods Mice were divided into 4 groups: the control group,the model group,the c-SKI gene infection group (LV-SKI group) and the normal control virus infection group (LV-NC group), with 10 in each. After two weeks, in the model group, isoprenaline(ISO) was injected subcutaneously for 30 d, with the first dose of 5 mg/kg followed by 2.5 mg/kg. In the LV-SKI group and the LV-NC group, c-SKI lentivirus and control virus were injected into the tail vein on days 18, 21, 24, 27 and 30 at a dose of 100 μL, titer was 1×10⁸ TU.HE staining and Masson staining was used to observe the pathological changes of myocardial tissue. ELISA was used to detect type Ⅰ and type Ⅲ collagen in myocardial tissue. Western blot was used to detect the expression of c-SKI, interstitial cell marker vimentin, α SMA,endothelial cell marker CD31 and the expression levels of transcription factors Snail, Twist and Slug. Results Compared with the control group, the expression of c-ski in the model group was time-dependent (P＜0.01). In the model group, the myocardial cells were disordered, the interstitium was significantly increased, and the contents of type Ⅰ and type Ⅲ collagen were increased (P＜0.01). Expressions of vimentin, α-SMA, Snail, Twist and Slug were all up regulated, while CD31 protein expression was down-regulated(P＜0.05). After over-expressing c-SKI, compared with the LV-NC group, the myocardial structure of the mice tended to be neat, the interstitial collagen fibers decreased, and the type Ⅰ and Ⅲ collagen in the myocardial tissue decreased (P＜0.01). The expression of vimentin and belt-sma was down-regulated (P＜0.05), and the expression of CD31 was up-regulated (P＜0.05). Meanwhile, the expression of transcription factors Snail, Twist and Slug was also down-regulated (P＜0.01). Conclusions c-SKI expression is down-regulated during myocardial fibrosis in mice, and over-expression of c-SKI can improve ISO-induced myocardial fibrosis by inhibiting End-MT.

Key words: c-SKI, cardiac fibrosis, endothelial-mesenchymal transition (End-MT), transcription factor

中图分类号:

张颖, 向燕, 张岳, 王娟. 过表达c-SKI抑制异丙肾上腺素诱导的小鼠心肌纤维化[J]. 基础医学与临床, 2021, 41(3): 364-369.

ZHANG Ying, XIANG Yan, ZHANG Yue, WANG Juan. Over-expression of c-SKI inhibits isoprenaline-induced myocardial fibrosis in mice[J]. Basic & Clinical Medicine, 2021, 41(3): 364-369.

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