左卡尼汀对心肌细胞纤维化增殖、凋亡及迁移的影响  

The effect of levocarnitine on fibrotic proliferation,apoptosis and migration of myocardial cells

作　　者：刘兆杰 金立 顾怡雯 石珏 汪海娅 方宁远[3] 舒锦 Liu Zhaojie;Jin Li;Gu Yiwen;Shi Jue;Wang Haiya;Fang Ningyuan;Shu Jin(Department of Geriatric Rehabilitation,Shibei Hospital,Jing'an District,Shanghai 200443,China;Department of Geriatrics,Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China;Department of Geriatrics,Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200001,China)

机构地区：[1]上海市静安区市北医院老年康复科,上海200443 [2]上海交通大学医学院附属第九人民医院老年科,上海200011 [3]上海交通大学医学院附属仁济医院老年科,上海200001

出　　处：《中华老年医学杂志》2024年第2期203-208,共6页Chinese Journal of Geriatrics

基　　金：上海市静安区医学重点学科建设项目(2021ZD03);上海市静安区卫生健康委员会科研课题(2020MS12);上海市卫生健康委员会中医药科研项目(2022QN006)。

摘　　要：目的探讨左卡尼汀对心肌细胞纤维化增殖、凋亡及迁移的影响作用机制。方法2022年6-12月,采用基质金属蛋白酶抑制剂-1(TIMP-1)过表达载体及siRNA-TIMP1转染大鼠心肌细胞H9c2(中国科学院细胞库),并用荧光定量反转录聚合酶链(RT-qPCR)检测转染效率。用血管紧张素Ⅱ(Ang Ⅱ)处理H9c2细胞后,通过RT-qPCR检测细胞中基质金属蛋白酶3(MMP3)、TIMP-1基因的表达情况。使用CCK8试剂盒检测TIMP-1过表达和敲低后,检测左卡尼汀干预对心肌细胞纤维化增殖的影响。通过流式凋亡检测和Transwell检测左卡尼汀对心肌细胞纤维化的凋亡及迁移影响。结果RT-qPCR结果表明AngⅡ处理24 h的心肌细胞中MMP3基因表达水平为(1.38±0.05),呈升高趋势(P<0.01),TIMP-1基因表达水平为(0.71±0.03),呈下调趋势(P<0.01);同时TIMP-1过表达为(905.98±24.17),敲低(0.18±0.01)%的H9c2细胞构建成功。CCK-8检测结果表明,TIMP-1敲低后表达量为(86.56±7.98)%可促进左卡尼汀诱导H9c2细胞增殖(P<0.01)。细胞凋亡实验结果表明,抑制TIMP-1表达为(23.22±2.69),抑制了左卡尼汀诱导的H9c2细胞凋亡水平(P<0.01)。迁移实验结果表明,抑制TIMP-1表达为(217.67±23.44)可促进左卡尼汀诱导的H9c2细胞迁移能力(P<0.01)。结论通过干预降低TIMP-1表达后,左卡尼汀可促进纤维化心肌细胞增殖,抑制纤维化心肌细胞凋亡,促进纤维化心肌细胞迁移,可能从而改善心肌纤维化。Objective To investigate the mechanisms underlying the effect of levocarnitine on myocardial cell fibrosis,proliferation,apoptosis and migration.MethodsBetween June and December 2022,an overexpression vector for tissue inhibitor-1 of metalloproteinase(TIMP-1)and siRNA TIMP-1 were used to transfect rat H9c2 cardiomyocytes(from the cell bank of the Chinese Academy of Sciences),and transfection efficiency was measured using fluorescence reverse transcription quantitative PCR(RT-qPCR).After treating H9c2 cells with angiotensin Ⅱ(Ang Ⅱ),the expression of the MMP3 and TIMP-1 genes in the cells was detected by RT-qPCR.A CCK8 kit was used to assess the effect of levocarnitine intervention on the proliferation of myofibroblasts after overexpression or knockdown of TIMP-1.The effects of levocarnitine on apoptosis and migration of myofibroblasts were detected by flow cytometry and Transwell assays.ResultsThe RT-qPCR results showed that the expression level of the MMP3 gene(1.38±0.05)in cardiomyocytes treated with AngⅡfor 24 hours exhibited an upward trend(P<0.01),while the expression level of the TIMP-1 gene(0.71±0.03)showed a downward trend(P<0.01).In addition,H9c2 cells with TIMP-1 overexpression(905.98±24.17)and knockdown(0.18±0.01)%,respectively,were successfully constructed.Based on CCK-8 detection results,knockdown of TIMP-1(86.56±7.98)%was able to promote the proliferation of H9c2 cells induced by levocarnitine(P<0.01).Apoptosis experiments showed that inhibition of TIMP-1 expression(23.22±2.69)significantly reduced the apoptosis level of H9c2 cells induced by levocarnitine(P<0.01).Migration experiments showed that inhibition of TIMP-1 expression(217.67±23.44)significantly promoted the migration ability of H9c2 cells induced by levocarnitine(P<0.01).ConclusionsAfter intervention to reduce TIMP-1 expression,levocarnitine can promote proliferation,inhibit apoptosis and promote migration of myofibroblasts and may therefore ameliorate myocardial fibrosis.

关 键 词：纤维化 肌细胞 心脏 基质金属蛋白酶类 

分 类 号：R96[医药卫生—药理学]