脑利钠肽对乳鼠心肌细胞肥大的抑制作用及可能机制  

Inhibitory effect and possible mechanism of brain natriuretic peptide on cardiomyocyte hypertrophy in suckling mice

作　　者：林杰 罗瑞英 吴露楠 LIN Jie;LUO Ruiying;WU Lunan(Department of Cardiovascular Medicine,2 Room of Medical Records,Ningde Municipal Hospital of Ningde Normal University,Ningde 352199,Fujian,China)

机构地区：[1]宁德师范学院附属宁德市医院心血管内科,福建省宁德市352199 [2]宁德师范学院附属宁德市医院病案室,福建省宁德市352199

出　　处：《广西医学》2023年第3期296-301,共6页Guangxi Medical Journal

基　　金：福建省卫生健康科技计划项目省医学创新课题(2020CXB045)。

摘　　要：目的分析脑利钠肽对苯肾上腺素(PE)诱导的乳鼠心肌细胞肥大的抑制作用,并基于过氧化物酶体增殖物激活受体δ(PPARδ)和脂肪酸氧化探讨其可能作用机制。方法(1)取新生昆明小鼠的左心室组织,制备心肌细胞。(2)将心肌细胞分为对照组、PE组、10 nmol/L脑利钠肽+PE组、100 nmol/L脑利钠肽+PE组和1000 nmol/L脑利钠肽+PE组。除对照组外,其余组心肌细胞均给予100μmol/L PE溶液处理,10 nmol/L脑利钠肽+PE组、100 nmol/L脑利钠肽+PE组和1000 nmol/L脑利钠肽+PE组细胞在PE处理前先分别加入相应浓度脑利钠肽溶液。对照组心肌细胞正常培养,不加任何药物。(3)将心肌细胞随机分为正常组、PE组、PE+抑制剂组和PE+脑利钠肽+抑制剂组。除正常组外,其余组心肌细胞给予100μmol/L PE溶液处理,PE+抑制剂组心肌细胞在PE处理前先加入1μmol/L GSK0660(PPARδ抑制剂)溶液处理,PE+脑利钠肽+抑制剂组心肌细胞在PE处理前同时加入1μmol/L GSK0660溶液和100 nmol/L脑利钠肽溶液处理。(4)干预48 h后,采用免疫荧光技术检测各组心肌细胞表面积;采用实时荧光定量PCR检测各组心肌细胞的心房钠尿肽(ANP)和β-肌球蛋白重链(β-MHC)mRNA表达水平;采用ELISA测定PPARδ抑制剂实验中各组心肌细胞的游离脂肪酸(FFA)含量;采用Western blot检测PPARδ抑制剂实验中各组心肌细胞的PPARδ、肉毒碱棕榈酰基转移酶1(CPT-1)和长链酰基辅酶A脱氢酶(LCAD)的蛋白表达水平。结果(1)与对照组比较,PE组心肌细胞ANP和β-MHC mRNA表达水平升高,心肌细胞表面积增大(均P<0.05);与PE组比较,各脑利钠肽干预组心肌细胞ANP和β-MHC mRNA表达水平降低,心肌细胞表面积减小(均P<0.05);与10 nmol/L脑利钠肽+PE组比较,100 nmol/L脑利钠肽+PE组、1000 nmol/L脑利钠肽+PE组心肌细胞ANP和β-MHC mRNA表达水平降低,心肌细胞表面积减小(均P<0.05);与100 nmol/L脑利钠肽+PE组比较,1000 nmol/L脑利钠肽+PE组�Objective To analyze the inhibitory effect of brain natriuretic peptide on cardiomyocyte hypertrophy in suckling mice induced by phenylephrine(PE),and to explore its possible mechanism based on peroxisome proliferator-activated receptorδ(PPARδ)and fatty acid oxidation.Methods(1)Left ventricular tissues of neonatal Kunming mice were obtained and prepared for cardiomyocytes.(2)Cardiomyocytes were assigned to control group,PE group,10 nmol/L brain natriuretic peptide+PE group,100 nmol/L brain natriuretic peptide+PE group,or 1000 nmol/L brain natriuretic peptide+PE group.Except for the control group,cardiomyocytes in the remaining groups received treatment of 100μmol/L PE solution,and brain natriuretic peptide solution with corresponding concentrations were added to the 10 nmol/L brain natriuretic peptide+PE group,the 100 nmol/L brain natriuretic peptide+PE group,and the 1000 nmol/L brain natriuretic peptide+PE group before PE treatment,respectively.Cardiomyocytes of the control group were normally cultured,without any medicine addition.(3)Cardiomyocytes were randomly assigned to normal group,PE group,PE+inhibitor group,or PE+brain natriuretic peptide+inhibitor group.Except for the normal group,cardiomyocytes in the remaining groups were treated with 100μmol/L PE solution,1μmol/L GSK0660(PPARδinhibitor)solution was added to cardiomyocytes in the PE+inhibitor group before PE treatment,and 1μmol/L GSK0660 solution and 100 nmol/L brain natriuretic peptide solution were simultaneously added to cardiomyocytes in the PE+brain natriuretic peptide+inhibitor group before PE treatment.(4)After 48 hours of intervention,the surface areas of cardiomyocytes were detected in various groups by the immunofluorescence technique.The mRNA expressions of atrial natriuretic peptide(ANP)and beta myosin heavy chain(β-MHC)were detected in cardiomyocytes in various groups by the real-time fluorescent quantitative PCR.The content of free fatty acid(FFA)of cardiomyocytes was measured in various groups in the PPARδinhibitor experiment b

关 键 词：心肌细胞肥大 脑利钠肽 过氧化物酶体增殖物激活受体Δ 脂肪酸氧化 乳鼠 细胞实验 

分 类 号：R542.2[医药卫生—心血管疾病]