过表达Trim72减轻急性病毒性心肌炎小鼠的心肌炎症及细胞凋亡  

Trim72 overexpression alleviates myocardial inflammation and apoptosis in mice with acute viral myocarditis

作　　者：王均祎 徐尚华 薛贻敏 柯俊[4] 章九云 谢辉 李珊 周晓芬[3] Wang Junyi;Xu Shanghua;Xue Yimin;Ke Jun;Zhang Jiuyun;Xie Hui;Li Shan;Zhou Xiaofen(Department of Intensive Care Unit,Nanping First Hospital Affiliated to Fujian Medical University,Nanping 353000,Fujian,China;Department of Cardiology,Nanping First Hospital Affiliated to Fujian Medical University,Nanping 353000,Fujian,China;Fourth Department of Critical Care Medicine,Shengli Clinical Medical College of Fujian Medical University,Fujian Provincial Hospital,Fuzhou University Affiliated Provincial Hospital,Fujian Provincial Key Laboratory of Emergency Medicine,Fuzhou 350001,Fujian,China;Department of Emergency,Shengli Clinical Medical College of Fujian Medical University,Fujian Provincial Hospital,Fuzhou University Affi liated Provincial Hospital,Fujian Provincial Key Laboratory of Emergency Medicine,Fuzhou 350001,Fujian,China)

机构地区：[1]福建医科大学附属南平第一医院重症医学科,南平353000 [2]福建医科大学附属南平第一医院心血管内科,南平353000 [3]福建医科大学省立临床医学院重症医学四科,福建省立医院,福州大学附属省立医院,福建省急诊医学重点实验室,福州350001 [4]福建医科大学省立临床医学院急诊科,福建省立医院,福州大学附属省立医院,福建省急诊医学重点实验室,福州350001

出　　处：《中华急诊医学杂志》2025年第2期193-199,共7页Chinese Journal of Emergency Medicine

基　　金：福建省自然科学基金项目(2022J01402,2023J011179);福建省卫生健康科研人才培养项目(2019-2-1)。

摘　　要：目的探讨三基序蛋白72(Trim72)在小鼠急性病毒性心肌炎(AVMC)中的作用及可能的机制。方法通过腹腔注射柯萨奇B3病毒(CVB3,2.0×105 PFU/只)法建立AVMC小鼠模型。40只小鼠按照随机数字表法分为阴性对照+磷酸缓冲盐溶液组(NC+PBS组)、过表达Trim72+PBS组(Trim72+PBS组)、NC+CVB3组和Trim72+CVB3组,每组10只。造模前14 d,各组小鼠分别经尾静脉注射阴性对照或过表达Trim72的9型腺相关病毒载体(5.0×1011 VG/只),后PBS组和CVB3组再分别经腹腔注射PBS或CVB3,7 d后处死存活小鼠并收集心脏和血清。HE染色观察心肌病理改变,TUNEL染色检查心肌细胞凋亡;RT-qPCR检测各组心肌组织Trim72和促炎细胞因子TNF-α、IL-6和IL-1β的mRNA表达;ELISA检测血清中cTnI、TNF-α、IL-6和IL-1β的蛋白水平;Western blot检测心肌组织Trim72、凋亡相关蛋白Bax、Bcl-2、Cleaved caspase-3、Caspase-3及TLR4、p-p65、p65的蛋白表达。结果与NC+PBS组相比,NC+CVB3组小鼠心肌组织中Trim72的蛋白及mRNA表达水平显著下调(P<0.05)。与NC+CVB3相比,过表达Trim72显著增加心肌组织Trim72的蛋白及mRNA表达(P<0.05),改善心肌炎症损伤,降低心肌细胞凋亡指数(P<0.05),并减少心肌和血清中TNF-α、IL-6和IL-1β的表达(P<0.05);此外,过表达Trim72还能下调心肌组织中Bax、Cleaved caspase-3/Caspase-3、TLR4和p-p65的蛋白表达,上调Bcl-2的蛋白表达(P<0.05)。NC+PBS组与Trim72+PBS组小鼠各项指标比较差异均无统计学意义(P>0.05)。结论Trim72过表达通过抑制心肌炎症损伤和凋亡失衡减轻小鼠AVMC,其机制可能与负调控TLR4/NF-κB信号通路有关。Objective To investigate the role and possible mechanism of tripartite motif-containing protein 72(Trim72)in acute viral myocarditis(AVMC)in mice.Methods A mouse model of AVMC was established by intraperitoneal injection of Coxsackievirus B3(CVB3,2.0×105 PFU/mouse).Forty mice were randomly divided into the negative control(NC)+phosphate-buffered saline(PBS)group(NC+PBS group),Trim72 overexpression+PBS group(Trim72+PBS group),NC+CVB3 group,and Trim72+CVB3 group(n=10).Fourteen days before modeling,mice in each group were injected with adeno-associated virus type 9 vector(AAV9)encoding either negative control or Trim72 overexpression(5.0×1011 VG/mouse)via tail vein.Subsequently,PBS or CVB3 was injected intraperitoneally in the PBS and CVB3 groups,respectively.After seven days,the surviving mice were euthanized,and the heart and serum samples were collected.HE and TUNEL staining were used to observe the cardiac pathological changes and cardiomyocyte apoptosis,respectively.The mRNA expression levels of Trim72 and pro-inflammatory cytokines TNF-α,IL-6,and IL-1βin myocardial tissues of each group were detected by RT-qPCR.The protein levels of cTnI,TNF-α,IL-6,and IL-1βin serum were detected by ELISA.The expression levels of Trim72,apoptosis-related proteins(Bax,Bcl-2,Cleaved caspase-3,Caspase-3),TLR4,p-p65,and p65 were detected by Western blot.Results The protein and mRNA expression levels of Trim72 in myocardial tissues of mice in the NC+CVB3 group were signifi cantly downregulated compared with those in the NC+PBS group(P<0.05).Compared with the NC+CVB3 group,Trim72 overexpression significantly increased the protein and mRNA expression of Trim72 in myocardial tissues(P<0.05),ameliorated myocardial inflammatory injury,decreased the apoptotic index of cardiomyocytes(P<0.05),and reduced the levels of pro-inflammation cytokines TNF-α,IL-6,and IL-1βin the myocardium and serum(P<0.05).Additionally,Trim72 overexpression also downregulated the protein expression of Bax,Cleaved caspase-3/Caspase-3,TLR4,and p-p65,and upr

关 键 词：Trim72 柯萨奇B3病毒 急性病毒性心肌炎 炎症 凋亡 TLR4/NF-κB 

分 类 号：R542.21[医药卫生—心血管疾病]