骨髓间充质干细胞来源外泌体对异丙肾上腺素诱导的大鼠心肌纤维化的影响及其作用机制  

Effect of bone marrow mesenchymal stem cell-derived exosomes on myocardial fibrosis in rats induced by isoproterenol and its mechanism

作　　者：魏俊萍 符达佳 孟庆雯 林道飞 林燕仔 WEI Junping;FU Dajia;MENG Qingwen;LIN Daofei;LIN Yanzai(Department of Cardiovascular,First Affiliated Hospital,Hainan Medical College,Haikou 570102,China;Echocardiography Room,First Affiliated Hospital,Hainan Medical College,Haikou 570102,China)

机构地区：[1]海南医学院第一附属医院心血管内一科,海南海口570102 [2]海南医学院第一附属医院心电图室,海南海口570102

出　　处：《吉林大学学报（医学版）》2024年第5期1348-1357,共10页Journal of Jilin University：Medicine Edition

基　　金：海南省卫健委卫生健康行业科研项目(22A200151);海南省科技厅自然科学基金青年项目(821QN401);海南医学院第一附属医院青年培育基金(HYYFYPY202020)。

摘　　要：目的:探讨骨髓间充质干细胞(BMSCs)来源外泌体(Exo)对异丙肾上腺素(ISO)诱导的大鼠心肌纤维化的影响,并初步阐明其作用机制。方法:从BMSCs中分离提取Exo,采用透射电子显微镜、纳米颗粒跟踪分析仪和Western blotting法对Exo进行分析鉴定。将40只SD大鼠分为对照组、模型组、BMSCs-Exo组和BMSCs-Exo+铁死亡激活剂(Erastin)组,每组10只。除对照组外,其余3组大鼠均皮下注射ISO建立心肌纤维化模型,BMSCs-Exo组和BMSCs-Exo+Erastin组大鼠尾静脉注射BMSCs-Exo,且BMSCs-Exo+Erastin组大鼠再腹腔注射Erastin。4周后,超声心动图检测各组大鼠左心室射血分数(LVEF)、左心室短轴缩短率(LVFS)、左心室舒张末期内径(LVEDD)和左心室收缩末期内径(LVESD), HE染色观察各组大鼠心肌组织病理形态表现,Masson染色观察各组大鼠心肌组织纤维化程度,免疫组织化学染色检测各组大鼠心肌组织中α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(COL-Ⅰ)和Ⅲ型胶原(COL-Ⅲ)阳性表达率,比色法测定各组大鼠心肌组织中铁离子(Fe^(2+))水平,Western blotting法检测各组大鼠心肌组织中酰基辅酶A合成酶长链家族成员4 (ACSL4)、铁蛋白重链1 (FTH1)、谷胱甘肽过氧化物酶4 (GPX4)和溶质载体家族7成员11 (SLC7A11)蛋白表达水平。结果:从BMSCs中分离的颗粒物具有典型的脂质双分子层,粒径大多分布在波长100 nm处,CD63、CD9、肿瘤易感基因101 (TSG101)和热休克蛋白70 (HSP70)蛋白均高表达,由此判定为Exo。与对照组比较,模型组大鼠LVEF和LVFS明显降低(P<0.05),LVEDD和LVESD明显升高(P<0.05),心肌细胞排列紊乱,部分细胞核皱缩和坏死,胶原容积分数(CVF)明显升高(P<0.05),α-SMA、COL-Ⅰ和COL-Ⅲ阳性表达率明显升高(P<0.05),心肌组织中Fe^(2+)水平明显升高(P<0.05),ACSL4蛋白表达水平明显升高(P<0.05),FTH1、GPX4和SLC7A11蛋白表达水平明显降低(P<0.05)。与模型组比较,BMSCs-Exo组大鼠LVEF和LVFS明显升�Objective:To discuss the effect of bone marrow mesenchymal stem cells(BMSCs)-derived exosomes(Exo)on isoproterenol(ISO)-induced myocardial fibrosis in the rats,and to clarify its mechanism.Methods:The Exo was isolated from the BMSCs and characterized by transmission electron microscope,nanoparticle tracking analysis,and Western blotting methods.Forty SD rats were divided into control group,model group,BMSCs-Exo group,and BMSCs-Exo+ferroptosis activator(Erastin)group,and there were 10 rats in each group.The myocardial fibrosis models were established by subcutaneous injection of ISO in all the rats except control group.The rats in BMSCs-Exo and BMSCs-Exo+Erastin groups were given BMSCs-Exo,and the rats in BMSCs-Exo+Erastin group were additionally injected with Erastin intraperitoneally.After 4 weeks,echocardiography was used to detect the left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),left ventricular end diastolic diameter(LVEDD),and left ventricular end systolic diameter(LVESD)of the rats in various groups;HE staining was used to observe the pathomorphology of myocardium tissue of the rats in various groups;Masson staining was used to observe the fibrosis degrees of myocardium tissue of the rats in various groups;immunohistochemistry was used to detect the positive expression rates ofα-smooth muscle actin(α-SMA),typeⅠcollagen(COL-Ⅰ),and typeⅢcollagen(COL-Ⅲ)in myocardium tissue of the rats in various groups;colorimetry was used to detect the Fe^(2+)level in myocardium tissue of the rats in various groups;Western blotting method was used to detect the expression levels of acyl-CoA synthetase long chain family member 4(ACSL4),ferritin heavy chain 1(FTH1),glutathione peroxidase 4(GPX4),and solute carrier family 7 member 11(SLC7A11)proteins in myocardium tissue of the rats in various groups.Results:The particles isolated from BMSCs had a typical lipid bilayer structure,and most particle sizes distributed around 100 nm.High expression levels of CD63,CD9,tumor susceptibi

关 键 词：心肌纤维化 异丙肾上腺素 骨髓间充质干细胞 外泌体 铁死亡 

分 类 号：R542.2[医药卫生—心血管疾病]