B10细胞减缓病毒性心肌炎诱导的心肌肥厚的机制研究  

IL-10 producing regulatory B cells ameliorated cardiac hypertrophy of viral myocarditis mice

作　　者：张士清 陈嘉 夏琳 苏兆亮[1,3] 许化溪[1] 刘芳 Zhang Shiqing;Chen Jia;Xia Lin;Su Zhaoliang;Xu Huaxi;Liu Fang(International Genome Center,Jiangsu University,Zhenjiang 212013,China;Department of Laboratory Medicine,Affiliated Hospital of Jiangsu University,Zhenjiang 212001,China;Department of Central Laboratory,The Fourth People′s Hospital of Zhenjiang,Zhenjiang 212001,China)

机构地区：[1]江苏大学国际基因组学研究中心,镇江212013 [2]江苏大学附属医院检验科,镇江212001 [3]镇江市第四人民医院中心实验室,212001

出　　处：《中华微生物学和免疫学杂志》2021年第5期345-352,共8页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金(81771756,81902136);中国博士后科学基金(2019M660106);江苏省重点研发计划(社会发展)项目(BE2019700);江苏省333工程科研项目资助计划(BRA2018016)。

摘　　要：目的:探讨心肌炎后B10细胞参与心肌细胞肥大的机制,为预防心肌炎诱导的心肌肥大探寻潜在的治疗策略。方法:体内实验选取柯萨奇病毒B3感染BALB/c小鼠诱导的病毒性心肌炎模型,检测心脏肥大相关指标,检测心肌炎小鼠血管紧张素Ⅱ及其受体表达量,流式分析对照组小鼠以及心肌炎小鼠心脏中B10细胞的改变。氯沙坦灌胃心肌炎小鼠后,苏木精-伊红染色检测心肌炎症程度,ELISA检测炎症因子表达,麦胚凝集素(WGA)染色检测心肌肥大,流式分析心脏中B10细胞改变情况。体外分别使用血管紧张素Ⅱ和血管紧张素Ⅱ+IL-10处理新生小鼠心肌细胞,检测肌钙蛋白T(C-TNT)和高迁移率族蛋白B1(HMGB1)水平。分别用血管紧张素Ⅱ、血管紧张素Ⅱ+B10细胞、血管紧张素Ⅱ+B10细胞+IL-10受体中和抗体和血管紧张素Ⅱ+B细胞处理心肌细胞,Annexin-V/PI检测心肌细胞凋亡,Western blot检测C-TNT蛋白水平。分别用氧化型HMGB1、还原型HMGB1和二硫键型HMGB1处理心肌细胞,检测C-TNT蛋白表达。结果:柯萨奇病毒B3感染引起小鼠心脏肥大,血管紧张素Ⅱ及其受体高表达,B10细胞一过性增多。氯沙坦处理阻断血管紧张素受体致B10细胞扩增,B10细胞通过产生IL-10减轻血管紧张素Ⅱ诱导的心肌细胞凋亡、抑制血管紧张素Ⅱ诱导的应激性心肌细胞HMGB1的产生,从而缓解病毒性心肌炎,减轻心肌肥大。结论:B10细胞缓解心肌炎诱导的心肌肥厚,在心肌保护中发挥重要作用。Objective To explore the mechanism of B10 cell involved in cardiomyocyte hypertrophy following myocarditis,and to develop potential therapeutic strategies.Methods BALB/c mice infected with Coxsackie virus B3 induced viral myocarditis model.The expression of angiotensin(ANG)Ⅱand its receptor in myocarditis mice was detected.The changes of B10 cells in the hearts of control mice and myocarditis mice were analyzed by flow cytometry.After losartan was administered to myocarditis mice,the degree of myocardial inflammation was detected by HE staining,the expression of inflammatory factors was detected by ELISA,the myocardial hypertrophy was detected by wheat germ agglutinin(WGA)staining,and the changes of B10 cells in the heart were analyzed by flow cytometry.The levels of cardiac troponin T(C-TNT)and high mobility group box 1(HMGB1)protein in neonatal mouse cardiomyocytes treated with ANGⅡand ANGⅡ+IL-10 were detected.Cardiomyocytes were treated with ANGⅡ,ANGⅡ+B10 cells,ANGⅡ+B10 cells+IL-10 receptor antibody and ANGⅡ+B cells to detect C-TNT protein levels,and Annexin-V/PI was used to detect the apoptosis of cardiomyocytes.Cardiomyocytes were treated with oxidized HMGB1,reduced HMGB1 and disulfide HMGB1,and C-TNT expression was detected.Results Coxsackievirus B3 infection caused cardiac hypertrophy,high expression of ANGⅡand its receptor,and transient increase of B10 cells in mice.Losartan treatment blocked the angiotensin receptor,reduced expansion of B10 cells.B10 cells alleviated apoptosis of cardiomyocytes and inhibited the production of HMGB1 induced by ANGⅡpatch by producing IL-10,thus alleviating viral myocarditis and cardiac hypertrophy.Conclusions B10 cells may play an important role in myocardial protection in myocarditis.

关 键 词：B10细胞 心脏重塑 心肌肥大 IL-10 HMGB1 

分 类 号：R542.21[医药卫生—心血管疾病]