回阳救急汤对慢性心力衰竭大鼠心肌细胞线粒体分裂/融合与凋亡影响的研究  

Effects of Huiyang Jiuji Decoction on Mitochondrial Fission/Fusion andApoptosis in Cardiomyocytes of Rats with Chronic Heart Failure

作　　者：张欢[1] 张莉[1] 辛玉 杨晨 岳文华 李立 殷可婧[1] ZHANG Huan;ZHANG Li;XIN Yu;YANG Chen;YUE Wenhua;LI Li;YIN Kejing(The Second Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China)

机构地区：[1]陕西中医药大学第二附属医院,陕西咸阳712000

出　　处：《中医药学报》2024年第2期30-36,共7页Acta Chinese Medicine and Pharmacology

基　　金：咸阳市重点研发计划(L2022ZDYFSF012);2019年全国中医药创新人才培养项目(国中医药人教函[2019]128号)。

摘　　要：目的:观察回阳救急汤(HYJJ)对慢性心力衰竭(CHF)大鼠心肌细胞线粒体分裂/融合与凋亡的影响。方法:24只大鼠随机分为空白组、模型组、回阳救急汤组和地高辛组,每组6只。大鼠腹腔注射盐酸阿霉素(ADMh, 1.5 mg/kg)建立CHF模型,每周2次,连续4周。建模成功后,将大鼠分为模型组、回阳救急汤组(给予回阳救急汤7.25 g/kg)和地高辛组(给予地高辛0.027 mg/kg);回阳救急汤组和地高辛组给予相应剂量药物灌胃,每日1次,连续2周;空白组和模型组给予等量生理盐水。ELISA法检测血清CK-MB、cTNⅠ、NT-proBNP水平;Tunel染色检测大鼠心肌组织细胞凋亡;免疫荧光双染Tom20和Drp1观察心肌细胞线粒体分裂;Western blot法检测大鼠Cleaved Caspase-3、Drp1、Fis1、Mfn1、OPA1蛋白表达水平;透射电镜观察大鼠心肌细胞线粒体结构。结果:与空白组相比,回阳救急汤组大鼠心肌细胞基本正常,未见明显病理变化;部分线粒体有轻度肿胀,可见少量自噬。与空白组相比,模型组大鼠cTNⅠ和NT-proBNP水平、心肌细胞凋亡率、Cleaved Caspase-3水平、Drp1、Tom20水平、Drp1、Fis1蛋白表达水平升高(P<0.01);Mfn1、OPA1蛋白表达水平降低(P<0.01)。与模型组相比,回阳救急汤组、地高辛组大鼠血清中cTNⅠ和NT-proBNP水平、心肌细胞凋亡率、Cleaved Caspase-3蛋白水平、Drp1、Tom20蛋白表达水平、Drp1、Fis1蛋白表达水平降低(P<0.05,P<0.01);Mfn1、OPA1蛋白表达水平升高(P<0.01)。结论:回阳救急汤抑制慢性心力衰竭大鼠心肌细胞凋亡,减缓心肌细胞线粒体分裂,促进线粒体融合。Objective:To observe the effects of Huiyang Jiuji(HYJJ)decoction on mitochondrial fission/fusion and apoptosis in cardiomyocytes of rats with chronic heart failure(CHF).Methods:Twenty-four rats were randomly divided into a blank group,a model(CHF)group,a Huiyang Jiuji(HYJJ)decoction group,and a digoxin group,with 6 rats in each group.Adriamycin hydrochloride(ADMh,1.5 mg/kg)were intraperitoneally injected into the rats twice a week for 4 weeks to establish CHF model.After successful modeling,the rats were divided into CHF,HYJJ(HYJJ decoction was administered 7.25 g/kg)and digoxin(digoxin was given 0.027 mg/kg)groups.HYJJ and digoxin groups were gavaged with corresponding dose of the drugs,once a day,for 2 weeks;while control and CHF groups were gavaged with the same amount of normal saline.Serum CK-MB,cTNⅠand NT-proBNP levels were detected through ELISA and Tunel staining was used to detect apoptosis of rat myocardial tissue cells.Mitochondrial division of myocardial cells was observed by immunofluorescence double staining Tom20 and Drp1.The levels of cleaved Caspase-3,Drp1,Fis1,Mfn1 and OPA1 were detected by Western blot,and mitochondrial structures of rat cardiomyocytes were observed by transmission electron microscopy.Results:Compared with blank group,the cardiomyocytes in HYJJ decoction group were basically normal,and no obvious pathological changes were observed.Some mitochondria showed mild swelling and a small amount of autophagy.Compared with blank group,cTNⅠand NT-proBNP levels,myocardial cell apoptosis rate,cleaved Caspase-3 levels,Drp1,Tom20 levels,Drp1 and Fis1 protein expression levels were increased in model group.The expression levels of Mfn1 and OPA1 proteins were decreased.Compared with model group,serum levels of cTNⅠand NT-proBNP,myocardial cell apoptosis rate,cleaved Caspase-3 protein level,Drp1 and Tom20 protein expression levels,Drp1 and Fis1 protein expression levels of rats in HYJJ decoction group and digoxin group were decreased(P<0.05,P<0.01),and the expression levels of Mfn1 and OP

关 键 词：回阳救急汤 慢性心力衰竭 线粒体 

分 类 号：R285.5[医药卫生—中药学]