沙库巴曲/缬沙坦对心力衰竭兔心肌细胞的保护作用及机制研究  

Protective effects and mechanism of sacubitril/valsartan on cardiomyocytes of rabbits with heart failure

作　　者：庄金龙[1,2] 钱涛铭 林庚海 陈华[2] 阮发晖 林惠萍[2] 刘莉 ZHUANG Jinlong;QIAN Taoming;LIN Genghai;CHEN Hua;RUAN Fahui;LIN Huiping;LIU Li(Heilongjiang University of Chinese Medicine,Harbin 150040,Heilongjiang,China;Department of Cardiovasology,Southeast Hospital Affiliated to Xiamen University,Zhangzhou 363000,Fujian,China;Department of Cardiovasology(Ⅰ),The First Affiliated Hospital,Heilongjiang University of Traditional Chinese Medicine,Harbin 150040,Heilongjiang,China)

机构地区：[1]黑龙江中医药大学,哈尔滨150040 [2]厦门大学附属东南医院心血管内科,漳州363000 [3]黑龙江中医药大学附属第一医院心血管病一科,哈尔滨150040

出　　处：《海军军医大学学报》2025年第3期360-373,共14页Academic Journal of Naval Medical University

基　　金：国家自然科学基金(82074346);福建省自然科学基金(2023J011835);漳州市自然科学基金(ZZ2018J14)。

摘　　要：目的探讨沙库巴曲(Sac)/缬沙坦(Val)对多柔比星(DOX)诱导的心力衰竭兔心肌细胞的保护作用及机制。方法选取30只新西兰兔建立DOX诱导的心力衰竭兔模型,25只造模成功,将造模成功的实验兔随机分为模型组(DOX组,9只)、Val干预组(DOX+Val组,8只)、Sac/Val干预组(DOX+Sac/Val组,8只);另取8只新西兰兔作为空白组。DOX+Val组每次灌胃Val混悬液4.65 mg/kg,DOX+Sac/Val组每次灌胃Sac/Val混悬液9.3 mg/kg,空白组及DOX组每次灌胃等体积的蒸馏水;各组均每日灌胃2次,连续8周。给药8周后,用超声心动图检测兔左心室舒张末期内径(LVDD)、左心室收缩末期内径(LVSD)、左心室射血分数(LVEF)、左心室短轴缩短率(LVFS);计算全心重量指数(HMI)及左心室重量指数(LVMI);通过H-E和马松染色观察心肌组织病理形态及纤维化情况;通过透射电镜观察心肌细胞超微结构;采用TUNEL检测观察心肌细胞凋亡情况,并计算心肌细胞凋亡率;采用ELISA法检测血清氨基末端脑利尿钠肽前体(NT-proBNP)、超敏肌钙蛋白I(Hs-cTNI)、血管紧张素Ⅱ(AngⅡ)、醛固酮(ALD)、心房利尿钠肽(ANP)、脑利尿钠肽(BNP)、环磷酸鸟苷(cGMP)、蛋白激酶G(PKG)水平;采用qPCR检测心肌组织利尿钠肽受体A(NPR-A)、cGMP特异性磷酸二酯酶5A[PDE5A(cGMP)]、PKG、Bcl-2、Bax、caspase 3 mRNA的表达;采用蛋白质印迹法检测心肌组织磷酸化cAMP反应元件结合蛋白(p-CREB)、磷酸化Bcl-2相关死亡促进因子(p-Bad)、Bcl-2、Bax、caspase 3蛋白的表达。结果与空白组相比,DOX组兔的LVDD、LVSD均增大(均P<0.01),LVEF、LVFS均降低(均P<0.01),HMI、LVMI均升高(均P<0.01);心肌细胞凋亡增加,心肌细胞凋亡率升高(P<0.01);NT-proBNP、Hs-cTNI、AngⅡ、ALD、ANP、BNP、cGMP、PKG水平和NPR-A、PDE5A(cGMP)、PKG、p-CREB、Bax、caspase 3表达均升高(均P<0.01),Bcl-2表达降低(P<0.01),p-Bad表达差异无统计学意义(P>0.05)。与DOX组相比,DOX+Sac/Val组和DOX+Val组的LVDD�Objective To study the protective effects and mechanism of sacubitril(Sac)/valsartan(Val)on cardiomyocytes of rabbits with heart failure induced by doxorubicin(DOX).Methods Thirty New Zealand rabbits were selected to establish DOX-induced heart failure rabbit model.Twenty-five rabbits with successful modeling were randomly assigned to model group(DOX group,n=9),DOX+Val group(n=8),and DOX+Sac/Val group(n=8);and another 8 New Zealand rabbits were selected as blank group.The DOX+Val group was gavaged with 4.65 mg/kg Val suspension each time,the DOX+Sac/Val group was gavaged with 9.3 mg/kg Sac/Val suspension each time,and the blank group and DOX group were gavaged with equal volume of distilled water each time.Each group was gavaged twice a day for 8 weeks.After 8 weeks of administration,echocardiography was used to measure left ventricular end-diastolic diameter(LVDD),left ventricular end-systolic diameter(LVSD),left ventricular ejection fraction(LVEF),and left ventricular fractional shortening(LVFS).The heart mass index(HMI)and left ventricular mass index(LVMI)were calculated.The pathological morphology and myocardial fibrosis of myocardial tissue were observed by hematoxylin-eosin(H-E)and Masson staining.The ultrastructure of cardiomyocytes was observed by transmission electron microscope.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining was used to observe cardiomyocytes apoptosis and apoptosis rate was calculated.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of N-terminal pro-brain natriuretic peptide(NT-proBNP),high-sensitivity cardiac troponin I(Hs-cTNI),angiotensinⅡ(AngⅡ),aldosterone(ALD),atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),cyclic guanosine monophosphate(cGMP),and protein kinase G(PKG)in serum.Quantitative polymerase chain reaction(qPCR)was used to detect the expression of natriuretic peptide receptor A(NPR-A),cGMP-specific phosphodiesterase 5A(PDE5A[cGMP]),PKG,B-cell lymphoma 2(Bcl-2),Bcl-2 associated X protein(Bax),and

关 键 词：沙库巴曲 缬沙坦 心力衰竭 心肌细胞 细胞凋亡 心功能 

分 类 号：R54[医药卫生—心血管疾病]