摘要
目的探讨小鼠心房肌细胞外泌体miR⁃130在房颤(AF)发病中的作用机制。方法用miR⁃130 shRNA慢病毒(sh⁃miR⁃130)或对照(sh⁃Crtl)转染小鼠心房肌细胞,然后进行AngⅡ处理。从处理过的心房肌细胞中分离出外泌体,与THP⁃1巨噬细胞共培养。进行流式细胞术以检查CD86和CD163阳性巨噬细胞的百分比。此外,通过构建横向主动脉弓缩窄术(TAC)小鼠模型,将sh⁃miR⁃130注射到小鼠体内以敲低miR⁃130表达,进一步研究miR⁃130在体内心脏纤维化过程和巨噬细胞极化中的作用。结果AngⅡ处理的心房肌细胞衍生的外泌体(AngⅡ⁃Exo)中miR⁃130表达上调。AngⅡ⁃Exo通过转移miR⁃130诱导巨噬细胞向M1极化。在有或没有sh⁃Ctrl对照处理的TAC小鼠中miR⁃130的水平显著升高,并且注射sh⁃miR⁃130的TAC小鼠的miR⁃130水平显著降低。sh⁃miR⁃130明显改善了TAC小鼠心脏的结构并减小了心肌大小、纤维化水平。此外,sh⁃miR⁃130处理后TAC小鼠左心房组织中iNOS^(+)/CD68^(+)巨噬细胞显著降低,和CD163^(+)/CD68^(+)巨噬细胞显著增加。结论在AngⅡ处理的心房肌细胞衍生的外泌体中miR⁃130的表达水平升高,并且外泌体通过转移miR⁃130促进了M1巨噬细胞极化。
Objective To explore the mechanism of exosomal miR-130 in mouse atrial myocytes in the pathogenesis of atrial fibrillation(AF).Methods Following AngⅡtreatment,mouse atrial myocytes were transfected with miR-130 shRNA lentivirus(sh-miR-130)or control(sh-Crtl).Exosomes were isolated from the treated atrial myocytes and then co-cultured with THP-1 macrophages.Flow cytometry was performed to examine the percentages of CD86-and CD163-positive macrophages.In addition,by constructing a transverse aortic arch coarctation(TAC)mouse model and injecting sh-miR-130 into mice to knock down miR-130 expression,the role of miR-130 in cardiac fibrosis and macrophages polarization in vivo was further studied.Results The expression of miR-130 was increased in AngⅡ-treated atrial myocyte-derived exosomes(AngII-Exo).By transferring miR-130,AngⅡ-Exo induced M1 polarization in macrophages.TAC mice with or without sh-Ctrl control treatment had significantly higher levels of miR-130,whereas TAC mice with sh-miR-130 injection had significantly lower levels.sh-miR-130 significantly improved cardiac structure and reduced myocardial size and fibrosis levels in TAC mice.In addition,iNOS^(+)/CD68^(+)macrophages had significantly decreased while CD163^(+)/CD68^(+)macrophages had significantly increased in left atrial tissue of TAC mice after sh-miR-130 treatment.Conclusion Exosomes from AngⅡ-treated atrial myocytes have higher levels of miR-130 expression,and exosomes promote M1 macrophage polarization by transferring miR-130.
作者
黄江伟
江鑫
蒋长荣
龚慧琴
齐明旭
HUANG Jiang-wei;JIANG Xin;JIANG Changrong;GONG Huiqin;QI Mingxu(Department of Cardiovascular Medicine,Nan-hua Hospital,University of South China,Hengyang 421002,China)
出处
《实用医学杂志》
CAS
北大核心
2022年第14期1766-1772,共7页
The Journal of Practical Medicine
基金
2020年度湖南省卫生健康委员会科研课题(编号:20200157)
衡阳市科技计划项目(编号:2019jb010920)。
