黄芩素对EAM模型小鼠心肌组织纤维化的影响  

Effect of baicalein on fibrosis of myocardial tissue in EAM model mice

作　　者：王甜甜[1] 王爽[1] 李玲[1] WANG Tiantian;WANG Shuang;LI Ling(School of Basic Medicine,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学基础医学院,山东青岛266071

出　　处：《精准医学杂志》2024年第2期95-100,107,共7页Journal of Precision Medicine

基　　金：国家自然科学基金资助项目(82001754)。

摘　　要：目的探讨黄芩素对自身免疫性心肌炎(experimental autoimmune myocarditis,EAM)模型小鼠心肌组织纤维化的影响及其机制。方法将20只BalB/C小鼠随机分为空白组、EAM组、黄芩素组、地塞米松组,每组5只。在饲养至第0、7天时,除空白组小鼠外余均皮下注射200μL完全弗氏佐剂与α-MyHC肽混合乳剂,饲养至第8天时开始对EAM组、黄芩素组及地塞米松组小鼠分别给予羧甲基纤维素钠、黄芩素、地塞米松灌胃,给药至第21天时行超声心动图检测各组小鼠左心室射血分数(LVEF)、缩短分数(LVFS)、舒张末期内径(LVIDd)及收缩末期内径(LVIDs)。第21天时将各组小鼠脱颈处死后取心脏组织进行HE染色和Masson染色;采用流式细胞术检测各组小鼠脾细胞中Th17/Treg细胞比例;采用实时荧光定量聚合酶链反应(RT-qPCR)和免疫印迹法检测各组小鼠心脏组织中的胶原Ⅰ(CollagenⅠ)、基质金属蛋白酶-1(MMP-1)、白细胞介素17A(IL-17A)、Treg转录因子(FOXP 3)及白细胞介素10(IL-10)的mRNA及蛋白表达水平。结果超声心动图显示,黄芩素组小鼠LVEF、LVFS显著高于EAM组(F=51.55、35.38,q=8.08、5.97,P<0.05),LVIDd、LVIDs显著低于EAM组(F=4.64、17.72,q=2.88、4.77,P<0.05)。HE及Masson染色结果显示,黄芩素组及地塞米松组小鼠心脏组织炎症细胞的浸润和胶原纤维的沉积较EAM组有所改善。流式细胞术检测结果显示,黄芩素组及地塞米松组小鼠脾细胞中Th17/Treg细胞比例显著低于EAM组(F=6.80,q=3.60、3.06,P<0.05)。RT-qPCR结果显示,黄芩素组小鼠心脏组织中CollagenⅠ、IL-17A mRNA水平显著低于EAM组(F=112.40、77.49,q=14.57、8.27,P<0.05),MMP-1、FOXP 3、IL-10 mRNA水平显著高于EAM组(F=19.05~144.60,q=5.37~11.21,P<0.05)。免疫印迹法检测结果显示,黄芩素组小鼠心脏组织中CollagenⅠ、IL-17A蛋白表达显著低于EAM组(F=13.70、13.97,q=4.72、5.64,P<0.05),MMP-1、FOXP3、IL-10蛋白表达显著高于EAM组(F=6.77~17.21,q=3.32~6Objective To explore the effect of baicalein on myocardial fibrosis in experimental autoimmune myocarditis(EAM)mice and its mechanism.Methods Twenty BalB/C mice were divided into blank group,EAM group,baicalein group,and dexamethasone group,with 5 mice in each group.On days 0 and 7,all the mice except those in the blank group were subcutaneously injected with 200μL of complete Freund’s adjuvant andα-MyHC peptide emulsion.Starting from day 8,the mice in the EAM group,baicalein group,and dexamethasone group were intragastrically administered sodium carboxymethyl cellulose,baicalein,and dexamethasone,respectively.Echocardiography was performed on day 21 of administration to determine left ventricular ejection fraction(LVEF),shortening fraction(LVFS),left ventricular end-diastolic internal diameter(LVIDd),and left ventricular end-systolic internal diameter(LVIDs)in mice across all groups.On day 21,the mice from all groups were euthanized by cervical dislocation and their heart tissues were collected for HE staining and Masson staining.Flow cytometry was used to detect the proportion of Th17/Treg cells among the splenocytes.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blot were used to analyze the mRNA and protein expression of CollagenⅠ,matrix metalloproteinase-1(MMP-1),interleukin 17A(IL-17A),Treg transcription factor(FOXP 3),and IL-10 in the cardiac tissue of mice.Results Echocardiographic results showed that the LVEF and LVFS of mice in baicalein group were significantly higher than those in the EAM group(F=51.55,35.38,q=8.08,5.97,P<0.05),while LVIDd and LVIDs were significantly lower than the EAM group(F=4.64,17.72,q=2.88,4.77,P<0.05).HE and Masson staining showed that infiltration of inflammatory cells and deposition of collagen fibers in the cardiac tissue of mice were significantly mitigated in the baicalein and dexamethasone groups as compared with the EAM group.Flow cytometry showed that the proportion of Th17/Treg cells was significantly higher in the EAM group than in the baicalein and

关 键 词：心肌炎 自身免疫疾病 纤维化 黄芩苷 TH17细胞 T淋巴细胞 调节性 基因表达调控 小鼠 

分 类 号：R542.21[医药卫生—心血管疾病] R593.2[医药卫生—内科学]