伊伐布雷定对病毒性心肌炎大鼠Bcl-2基因启动子区甲基化及mRNA表达的影响  

Effect of Ivabradine on Methylation of Bcl-2 Gene Promoter Region and mRNA Expression in Rats with Viral Myocarditis

作　　者：陈小霞 程齐尧 詹磊 蒋磊 CHEN Xiaoxia;CHENG Qiyao;ZHAN Lei;JIANG Lei(Anhui No.2 Provincial People′s Hospital,Hefei 230000,Anhui,China)

机构地区：[1]安徽省第二人民医院,合肥230000 [2]安徽医科大学第二附属医院

出　　处：《中西医结合心脑血管病杂志》2021年第12期1991-1995,共5页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：国家自然科学基金项目(No.81802586)。

摘　　要：目的探讨伊伐布雷定对病毒性心肌炎大鼠Bcl-2基因启动子区甲基化及mRNA表达的影响。方法将SPF级雄性SD大鼠随机分为对照组、模型组、伊伐布雷定低剂量组(4 mg/kg)、伊伐布雷定中剂量组(8 mg/kg)、伊伐布雷定高剂量组(12 mg/kg)。除对照组外,其余各组大鼠均采用腹腔接种柯萨奇病毒制备病毒性心肌炎大鼠模型,首次接种柯萨奇病毒后24 h给予药物干预,每日1次,连续干预2周,观察各组大鼠心肌功能。苏木精-伊红(HE)染色观察心肌组织细胞病理变化,脱氧核苷酸转移酶标记技术(TUNEL)检测心肌组织细胞凋亡情况,实时荧光定量聚合酶链反应(PCR)检测凋亡相关基因mRNA表达情况,甲基化特异性PCR(MSP)检测Bcl-2启动子区甲基化水平。结果模型组肌酸激酶同工酶(CK-MB)、肌红蛋白(Mb)、心肌肌钙蛋白I(cTnI)和乳酸脱氢酶(LDH)水平高于对照组(P<0.05);伊伐布雷定低剂量组、中剂量组、高剂量组CK-MB、Mb、cTnI和LDH水平低于模型组(P<0.05)。HE染色结果显示:模型组心肌组织存在明显炎性浸润、细胞坏死;伊伐布雷定低剂量组、中剂量组、高剂量组心肌组织损伤较模型组明显改善。模型组心肌组织细胞凋亡指数、Bax mRNA、CytC mRNA水平高于对照组(P<0.05),Bcl-2 mRNA水平低于对照组(P<0.05);伊伐布雷定低剂量组、中剂量组、高剂量组心肌组织细胞凋亡指数、Bax mRNA、CytC mRNA水平低于模型组(P<0.05),Bcl-2 mRNA水平高于模型组(P<0.05)。MSP检测结果显示:模型组Bcl-2基因启动子区DNA甲基化水平高于对照组(P<0.05);伊伐布雷定低剂量组、中剂量组、高剂量组Bcl-2基因启动子区DNA甲基化水平低于模型组(P<0.05)。结论伊伐布雷定可缓解病毒性心肌炎大鼠心肌组织细胞凋亡,可能与降低Bcl-2基因启动子区DNA甲基化有关。Objective To explore the effect of ivabradine on methylation of Bcl-2 gene promoter region and mRNA expression in rats with viral myocarditis.Methods The SPF male SD rats were randomly divided into control group,model group,low-dose ivabradine group(4 mg/kg),medium-dose ivabradine group(8 mg/kg),and high-dose ivabradine group(12 mg/kg).Except for the control group,rats in the other groups were intraperitoneally inoculated with Coxsackie virus to prepare the rat models of viral myocarditis.At 24 h after first Coxsackie virus inoculation,the rats were treated with drug intervention,once a day,for 2 weeks.The myocardial function of rats in each group was observed.The pathological changes of myocardial tissue cells were observed by hematoxylin-eosin(HE)staining.The apoptosis of myocardial tissue was detected by TUNEL,the mRNA expression of apoptosis-related genes was detected by real-time fluorescence quantitative PCR,and the methylation level of Bcl-2 promoter region was detected by methylation specific PCR(MSP).Results The levels of creatine kinase MB(CK-MB),myohemoglobin(Mb),cardiac troponin I(cTnI)and lactate dehydrogenase(LDH)in the model group were significantly increased than those in the control group(P<0.05).The levels of CK-MB,Mb,cTnI,and LDH in the low-dose ivabradine group,medium-dose ivabradine group,and high-dose ivabradine group significantly decreased than those in the model group(P<0.05).The results of HE staining showed that there were obvious inflammatory infiltration and cell necrosis in myocardial tissues of model group.Compared with the model group,the damage of myocardial tissue in the low-dose ivabradine group,medium-dose ivabradine group,and high-dose ivabradine group was significantly improved.The apoptosis index of myocardial tissue cells,Bax mRNA,and CytC mRNA in the model group significantly increased more than that did in control group(P<0.05),while Bcl-2 mRNA significantly decreased than that did in the control group(P<0.05).The apoptosis index of myocardial tissue cells,Bax mRNA and Cy

关 键 词：病毒性心肌炎 伊伐布雷定 心肌细胞凋亡 Bcl-2 甲基化 大鼠 实验研究 

分 类 号：R73[医药卫生—肿瘤]