[1]杨　莉,贺　静,孙晓慧,等.MicroRNA-185对血管紧张素Ⅱ介导的心肌细胞肥大和凋亡的影响[J].新乡医学院学报,2019,36(11):1024-1029.[doi:10.7683/xxyxyxb.2019.11.005]
　YANG Li,HE Jing,SUN Xiao-hui,et al.Effect of microRNA-185 on angiotensin Ⅱ mediated cardiomyocyte hypertrophy and apoptosis[J].Journal of Xinxiang Medical University,2019,36(11):1024-1029.[doi:10.7683/xxyxyxb.2019.11.005]
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MicroRNA-185对血管紧张素Ⅱ介导的心肌细胞肥大和凋亡的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
36
期数:
2019年11
页码:
1024-1029
栏目:
基础研究
出版日期:
2019-11-05

文章信息/Info

Title:
Effect of microRNA-185 on angiotensin Ⅱ mediated cardiomyocyte hypertrophy and apoptosis
作者:
杨　莉1; 贺　静1; 孙晓慧1; 乌宇亮2
（1.泾河工业园长庆油田职工医院心血管内科，陕西　西安　710201；2.西安交通大学第一附属医院心血管内科，陕西　西安　710061）
Author(s):
YANG Li1; HE Jing1; SUN Xiao-hui1; WU Yu-liang2
（1.Department of Cardiovascular Medicine,Jinghe Industrial Park Changqing Oilfield Worker′s Hospital,Xi′an 710201，Shaanxi Province，China；2.Department of Cardiovascular Medicine,the First Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710061,Shaanxi Province，China ）
关键词:
microRNA-185; 血管紧张素Ⅱ; 心肌肥大; 细胞凋亡; 细胞分裂周期蛋白42
Keywords:
microRNA-185; angiotensin Ⅱ; cardiac hypertrophy; cell apoptosis; cell division cycle 42
分类号:
R542.2
DOI:
10.7683/xxyxyxb.2019.11.005
文献标志码:
A
摘要:
目的　探讨MicroRNA-185（miR-185）对血管紧张素Ⅱ（Ang Ⅱ）介导的心肌细胞肥大及凋亡的影响。方法　将心肌细胞系H9c2细胞随机分为对照组、Ang Ⅱ处理组、阴性对照组和miR-185过表达组。对照组细胞采用常规培养；Ang Ⅱ处理组细胞常规培养，并给予Ang Ⅱ处理；阴性对照组细胞使用含miR-185阴性对照（NC）的无血清培养基处理24 h后给予Ang Ⅱ处理；miR-185过表达组细胞使用含miR-185模拟物（miR-185 mimic）的无血清培养基培处理24 h后给予Ang Ⅱ处理。采用实时定量聚合酶链反应检测各组细胞中miR-185、心房钠尿肽（ANP）、脑钠肽（BNP）、β-肌球蛋白重链（β-MHC）和细胞分裂周期蛋白42（CDC42） mRNA表达，应用细胞计数试剂盒-8检测各组细胞活性，细胞凋亡检测试剂盒测定各组细胞凋亡小体富计因子水平，Western blot法测定各组细胞中活化型多聚腺苷二磷酸核糖聚合梅（cleaved PARP）、活化型caspase 3和CDC42蛋白表达。结果　与对照组比较，Ang Ⅱ处理组和阴性对照组细胞中miR-185相对表达量、细胞活性显著降低（P<0.05），ANP、BNP、β-MHC和CDC42 mRNA相对表达量、细胞凋亡小体富计因子及活化型PARP、活化型 caspase 3和CDC42蛋白相对表达量显著升高（P<0.05）。阴性对照组与Ang Ⅱ处理组细胞中miR-185和ANP、BNP、β-MHC、CDC42 mRNA相对表达量、细胞活性、凋亡小体富计因子及活化型 PARP、活化型caspase 3、CDC42蛋白相对表达量比较差异均无统计学意义（P>0.05）。与Ang Ⅱ组和阴性对照组比较，miR-185过表达组细胞中miR-185相对表达量、细胞活性显著升高（P<0.05），ANP、BNP、β-MHC和CDC42 mRNA相对表达量、细胞凋亡小体富计因子及活化型PARP、活化型caspase 3、CDC42蛋白相对表达量显著降低（P<0.05）。结论　miR-185可能通过下调心肌细胞中CDC42水平来减轻Ang Ⅱ介导的心肌细胞损伤，抑制Ang Ⅱ介导的心肌细胞肥大和凋亡，改善心肌细胞活性，从而发挥心肌保护作用。
Abstract:
Objective　 To study the effect of microRNA-185（miR-185） on angiotensin Ⅱ (Ang Ⅱ)-mediated cardiomyocyte hypertrophy and apoptosis。Methods　Myocardial cell line H9c2 cells were randomly divided into control group,Ang Ⅱ group,negative control group and miR-185 overexpression group.The cells in the control group were cultured conventionally；the cells in the Ang Ⅱ group were treated with 0.1 μmol·L-1 Ang Ⅱ for 24 hours；the cells in the negative control group were treated with Ang Ⅱ after transfection with serum-free medium containing miR-185 negative control for 24 hours,and cells in the miR-185 overexpression group were treated with Ang Ⅱ after transfection with serum-free medium containing miR-185 mimic for 24 hours.The mRNA levels of atrial natriuretic peptide (ANP),brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC) and cell division cyclin 42 (CDC42) were detected by real time-quantitative polymerase chain reaction (RT-qPCR).The cell viability was detected by cell counting kit-8.The levels of apoptosis cell body enrichment factors were detected by cell apoptosis detection kit.The expressions of cleaved poly ADP-ribose polymerase (PARP),cleaved caspase 3 and CDC42 protein were determined by Western blot.Results　Compared with the control group，in the Ang Ⅱ group and negative control group,the expression of miR-185 and cell viability were significantly decreased (P<0.05),the mRNA levels of ANP,BNP,β-MHC and CDC42,apoptotic enrichment factors,and the expressions of cleaved PARP,cleaved caspase 3 and CDC42 protein were obviously increased (P<0.05).There was no significant difference in the miR-185 level,the mRNA level of ANP,BNP,β-MHC and CDC42,cell viability,apoptotic rich factor，and the expression of cleaved PARP,cleaved caspase 3 and CDC42 protein between the Ang Ⅱ group and negative control group (P>0.05).Compared with the Ang Ⅱ group and negative control group,the level of miR-185 and cell viability were significantly increased (P<0.05),and the expression of ANP,BNP,β-MHC and CDC42 mRNA,apoptotic rich factors,the expression of cleaved PARP,cleaved caspase 3 and CDC42 protein in the miR-185 overexpression group were significantly decreased in the miR-185 overexpression group (P<0.05).Conclusion　miR-185 may reduce Ang Ⅱ-mediated cardiomyocyte injury，inhibit Ang Ⅱ-mediated cardiomyocyte hypertrophy，improve cardiomyocyte activity，inhibit cardiomyocyte apoptosis，and play a role in myocardial protection by down-regulating the CDC42 level of cardiomyocytes.

参考文献/References:

［1］　MAKAVOS G,KAPPAAIRIS C,TSELEGKIDI M E,et al.Hypertrophic cardiomyopathy:an updated review on diagnosis,prognosis,and treatment［J］.Heart Fail Revi，2019，24(4)：439-459.
［2］　PINTI M V,HATHAWAY Q A,HOLLANDER J M.Role of microRNA in metabolic shift during heart failure［J］.Am J Physiol Heart Circ Physiol，2017，312(1)：H33-H45.
［3］　CHENG Y,JI R,YUE J,et al.MicroRNAs are aberrantly expressed in hypertrophic heart:do they play a role in cardiac hypertrophy［J］.Am J Pathol，2007，170(6)：1831-1840.
［4］　GESMUNDO I,MIRAGOLI M,CARULLO P,et al.Growth hormonereleasing hormone attenuates cardiac hypertrophy and improves heart function in pressure overload-induced heart failure［J］.Proc Natl Acad Sci USA,2017,114(45):12033-12038.
［5］　王卓,司澳洋,刘亚坤,等.抵抗素通过酪氨酸蛋白激酶2/信号转导子和转录激活子3信号通路诱导心肌细胞肥厚的分子机制研究［J］.新乡医学院学报，2017，34(4)：265-269.
［6］　肖坤.心肌肥厚的发病机制与治疗现状［J］.赣南医学院学报,2017,37(3):495-500.
［7］　SHENG Z,XU Y,LI F,et al.CSN5 attenuates Ang Ⅱ-induced cardiac hypertrophy through stabilizing LKB1［J］.Exp Cell Res,2019,376(1):11-17.
［8］　TANAKA Y,OBATA K,OHMORI T,et al.Angiotensin Ⅱ induces automatic activity of the isolated guinea pig pulmonary vein myocardium through activation of the IP3 receptor and the Na+-Ca2+ exchanger［J］.Int J Mol Sci,2019,20(7)：E1768.
［9］　YU X J,HUANG Y Q,SHAN Z X,et al.MicroRNA-92b-3p suppresses angiotensin Ⅱ-induced cardiomyocyte hypertrophy via targeting HAND2［J］.Life Sci，2019，232:116635.
［10］　DONG Z X,WAN L,WANG R J,et al.(-)-Epicatechin suppresses angiotensin Ⅱ-induced cardiac hypertrophy via the activation of the SP1/SIRT1 signaling pathway［J］.Cell Physio Biochem，2017，41(5)：2004-2015.
［11］　PARIZADEH S M,JAFARZADEH-ESFEHANI R,GHANDEHARI M,et al.Circulating and tissue microRNAs as biomarkers for ovarian cancer prognosis［J］.Curr Drug Targets，2019,20(14):1447-1460.
［12］　CHENG J Z,CHEN J J,WANG Z G,et al.MicroRNA-185 inhibits cell proliferation while promoting apoptosis and autophagy through negative regulation of TGF-β/mTOR axis and HOXC6 in nasopharyngeal carcinoma［J］.Cancer Biomark，2018，23(1)：107-123.
［13］　LI L,WANG X,LIU D.MicroRNA-185 inhibits proliferation,migration and invasion in human osteosarcoma MG63 cells by targeting vesicle-associated membrane protein 2［J］.Gene，2019，696：80-87.
［14］　JIANG X,NING Q,WANG J.Angiotensin Ⅱ induced differentially expressed microRNAs in adult rat cardiac fibroblasts［J］.J Physiol Sci，2013，63(1)：31-38.
［15］　WANG X,YANG C,LIU X,et al.Ghrelin alleviates angiotensin Ⅱ-induced H9c2 apoptosis:impact of the miR-208 family［J］.Med Sci Monit,2018，24：6707-6716.
［16］　柳培雨,田毅.血管紧张素转换酶抑制剂与心肌缺血再灌注的细胞凋亡［J］.医学与哲学，2010,31(9):41-43.
［17］　ZHANG D,LEE H,CAO Y,et al.miR-185 mediates lung epithelial cell death after oxidative stress［J］.Am J Physiol Lung Cell Mol Physiol，2016，310(7)：700-710.
［18］　KIM J O,KWON E J,SONG D W,et al.miR-185 inhibits endoplasmic reticulum stress-induced apoptosis by targeting Na+/H+exchanger-1 in the heart［J］.BMB Rep，2016，49(4)：208-213.
［19］　NAGATA S.Apoptosis and clearance of apoptotic cells［J］.Annual Rev Immunol,2018,36(1):489-517.
［20］　WEI Q,DENG H,CUI H,et al.A mini review of fluoride-induced apoptotic pathways［J］.Environ Sci Pollut Res Int，2018，25(34):33926-33935.
［21］　MAILLET M,LYNCH J M,SANNA B,et al.Cdc42 is an antihypertrophic molecular switch in the mouse heart［J］.J Clin Invest，2009，119(10)：3079-3088.
［22］　LIU Y,WANG J,LI J,et al.Deletion of Cdc42 in embryonic cardiomyocytes results in right ventricle hypoplasia［J］.Clin Transl Med，2017，6(1)：40.
［23］　XU X,KONG L,SONG X,et al.Effect of Cdc42 on myocardial ischemia-reperfusion of rats［J］.Cell Mol Biology,2017,63(7):31-34.

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更新日期/Last Update: 2019-11-05