目的探究黄芪甲苷(AS-Ⅳ)对柯萨奇B组3型病毒性心肌炎的保护作用及其机制,采用生物信息学分析方法进一步探索AS-Ⅳ发挥作用的潜在机制。方法60只8周龄雄性Balb/c小鼠,随机分为四组,即空白对照组(对照组)、病毒性心肌炎模型组(模型组)、AS-Ⅳ干预低剂量组(低剂量组)及AS-Ⅳ干预高剂量组(高剂量组),每组15只小鼠。低剂量组和高剂量组小鼠在病毒注射前使用AS-Ⅳ对小鼠进行预处理2周,根据实验设计,AS-Ⅳ的给药剂量分别为10、50 mg/kg,灌胃体积为3 ml/只。对照组和模型组小鼠灌胃相同体积的空白溶媒作为预处理。预处理结束后,模型组、低剂量组和高剂量组小鼠通过腹腔注射0.1 ml 103TCID50的柯萨奇B组3型病毒诱导病毒性心肌炎,而对照组小鼠腹腔注射相同体积的空白细胞悬液。病毒注射后,低剂量组和高剂量组小鼠继续进行AS-Ⅳ干预直到2周实验结束,在实验过程中每3天检测小鼠的体重和死亡情况。2周后处死小鼠,经过2周生存观察后收集外周血并分离血清,利用全自动生化分析仪检测心肌标志物水平;获得心脏组织进行分子生物学和病理学检测;使用苏木精-伊红染色法(HE)染色及Masson染色评估心肌损伤和纤维化水平;利用凋亡检测试剂盒检测心肌组织凋亡情况。然后利用生物数据库检索AS-Ⅳ干预与病毒性心肌炎相关靶点。使用string数据库及Cytoscape软件药物-靶点-疾病”网络分析,药物及疾病靶点基因取交集后使用metascape数据库进行Gene Ontology(GO)及Kyoto Encyclopedia of Genes and Genomes(KEGG)通路富集分析,研究黄芪有效成分保护病毒性心肌炎的潜在分子机制。结果截至到干预后2周,对照组小鼠全部存活(15/15,100%),模型组存活6只(6/15,40%),低剂量组存活7只(7/15,47%),高剂量组存活12只(12/15,80%),四组存活率比较有统计学差异(P<0.05),其中对照组存活率高于模型组、低剂量组,有统�
Objective:To explore the molecular mechanism of Shenmai Injection(SMI)against doxorubicin(DOX)induced cardiomyocyte apoptosis.Methods:A total of 40 specific pathogen-free(SPF)male Sprague Dawley(SD)male rats were divided into 5 groups based on the random number table,including the control group,the model group,miR-30a agomir group,SMI low-dose(SMI-L)group,and SMI high-dose(SMI-H)group,with 8 rats in each group.Except for the control group,the rats were injected weekly with DOX(2 mg/kg)in the tail vein for 4 weeks to induce myocardial injury,and were given different regimens of continuous intervention for 2 weeks.Cardiac function was detected by echocardiography and myocardial pathological changes were observed by Van Gieson(VG)staining.Myocardial injury serum markers,including creatine kinase(CK),lactate dehydrogenase(LDH),troponin T(c Tn T),N-terminal pro-brain natriuretic peptide(NT-pro BNP),soluble ST2(sST2),and growth differentiation factor-15(GDF-15)were detected by enzyme linked immunosorbent assay(ELISA).Cardiomyocyte apoptosis was observed by terminal deoxynucleotidyl transferase-mediated biotinylated d UTP triphosphate nick end labeling(TUNEL)and transmission electron microscopy,and the expressions of target proteins and m RNA were detected by Western blot and quantitative real time polymerase chain reaction(qRT-RCR),respectively.Results:The treatment with different doses of SMI reduced rat heart mass index and left ventricular mass index(P<0.05),significantly improved the left ventricular ejection fraction(P<0.05),decreased the levels of serum CK,LDH,cTnT,and NT-proBNP(P<0.05 or P<0.01),reduced the levels of serum sST2 and GDF-15(P<0.05 or P<0.01),decreased the collagen volume fraction,reduced the expressions of rat myocardial typeⅠand typeⅢcollagen(P<0.05 or P<0.01),and effectively alleviated myocardial fibrosis.And the study found that SMI promoted the expression levels of miR-30a and Bcl-2 in myocardium,and down-regulated the expression of Bax,which inhibited the activation of Caspase-3 and Caspa
