[1]刘永强,韩培立,刘　辉.神经调节蛋白-1原核表达及其对阿霉素致大鼠心肌细胞毒性的保护作用[J].新乡医学院学报,2020,37(2):101-106.[doi:10.7683/xxyxyxb.2020.02.001]
　LIU Yongqiang,HAN Peili,LIU Hui.Prokaryotic expression of neuregulin-1 and its protective effect on doxorubicin-induced cardiomyocytes toxicity in rats[J].Journal of Xinxiang Medical University,2020,37(2):101-106.[doi:10.7683/xxyxyxb.2020.02.001]
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神经调节蛋白-1原核表达及其对阿霉素致大鼠心肌细胞毒性的保护作用
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
37
期数:
2020年2
页码:
101-106
栏目:
基础研究
出版日期:
2020-02-05

文章信息/Info

Title:
Prokaryotic expression of neuregulin-1 and its protective effect on doxorubicin-induced cardiomyocytes toxicity in rats
作者:
刘永强1; 韩培立1; 刘　辉2
(1.新乡医学院第一附属医院心外科，河南省神经修复重点实验室，河南　卫辉　453100；2.新乡医学院第一附属医院心内科，河南　卫辉　453100)
Author(s):
LIU Yongqiang1; HAN Peili1; LIU Hui2
（1.Department of Cardiovascular Surgery，the First Affiliated Hospital of Xinxiang Medical University，Henan Key Laboratory of Neural Rehabilitation，Weihui 453100，Henan Province，China；2.Department of Cardiology，the First Affiliated Hospital of Xinxiang Medical University，Weihui 453100，Henan Province，China）
关键词:
神经调节蛋白-1; 阿霉素; 心肌球蛋白轻链激酶; Na+-Ca2+交换体
Keywords:
neuregulin-1; doxorubicin; cardiacmyosin light chain kinase; Na+-Ca2+exchanger
分类号:
R965；Q816
DOI:
10.7683/xxyxyxb.2020.02.001
文献标志码:
A
摘要:
目的　探讨神经调节蛋白-1(NRG-1）对减轻阿霉素（DOX）所致大鼠心肌细胞毒性的作用及其机制。方法　提取Sprague Dawley(SD)胎鼠的心室肌细胞总RNA并原核表达NRG-1蛋白；分离并培养SD乳鼠原代心肌细胞，应用细胞计数试剂盒(CCK-8)测定不同浓度DOX作用下大鼠原代心肌细胞的活性；并将心肌细胞分为正常对照组、DOX（5.0 μmol· L-1）组、DOX（5.0 μmol· L-1）+NRG-1（11.0 nmol· L-1）组和NRG-1（11.0 nmol· L-1）组，培养7 d后分别采用Western blot法和荧光定量聚合酶链反应检测大鼠原代心肌细胞的Na+-Ca2+交换体（NCX-1）和心肌球蛋白轻链激酶（cMLCK）蛋白和mRNA表达。结果　原核表达NRG-1蛋白成功；DOX的心肌细胞毒性随其浓度增高而加重；NRG-1对不同浓度DOX干预的大鼠心肌细胞活力均有恢复作用（P<0.05），DOX浓度继续增高时其恢复作用受限（P<0.05），同时NRG-1浓度越高其细胞活力恢复越好（F=3 606.28、2 010.60、215.41，P<0.05）；5.0 μmol· L-1 DOX能抑制cMLCK蛋白和mRNA的表达（P<0.05），也能提高NCX-1蛋白和mRNA的表达（P<0.05），而11.0 nmol· L-1 NRG-1能够逆转DOX的上述作用（P<0.05）。结论　NRG-1能够改善DOX所致大鼠心肌细胞毒性，其机制可能与cMLCK表达上调及NCX-1表达下调有关。
Abstract:
Objective　To investigate the effect of neuregulin-1（NRG-1）on reducing cardiomyocyte toxicity of rats induced by doxorubicin (DOX) and its mechanism.Methods　Total RNA was extracted from ventricular myocytes of Sprague Dawley(SD) fetal rats and NRG-1 was expressed in prokaryotic cells.Primary cardiomyocytes of SD rats were isolated and cultured.The cell counting kit-8（CCK-8）assay was used for determining the activity of rat cardiomyocytes at different DOX concentrations.Cardiomyocytes were divided into normal control group,DOX （5.0 μmol·L-1）group,DOX（5.0 μmol· L-1）+ NRG-1（11.0 nmol· L-1） group,and NRG-1（11.0 nmol· L-1） group.All the cells in each group were cultured for seven days.The protein expression of Na+-Ca2+exchanger（NCX-1）and cardiac myosin light chain kinase（cMLCK）was detected by Western blot,and the mRNA expression level was measured by fluorescence quantitative polymerase chain reaction.Results　NRG-1 was obtained by prokaryotic expression.DOX-induced cardiomyocytes toxicity became more significant with the increasing dose of DOX.The NRG-1 could restore the viability of cardiomyocytes under different does of DOX（P<0.05），However,the protective effect of NRG-1 was limited when the DOX concentration continued to incresase（P<0.05）;meanwhile,the cell viability recovery became more significant with higher degree of NRG-1 concentrations（F=3 606.28，2 010.60，215.41；P<0.05）.The 5.0 μmol· L-1 DOX could inhibit the expression of cMLCK protein and mRNA（P<0.05）,and increase the expression of NCX-1 protein and mRNA（P<0.05）,while the 11.0 nmol· L-1 NRG-1 could reverse the above effects of DOX (P<0.05).Conclusions　NRG-1 can improve DOX-induced cardiomyocyte toxicity in rats,and its mechanism may be related to the up-regulation of the cMLCK expression and the down-regulation of the NCX-1 expression.

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更新日期/Last Update: 2020-02-05