脑利钠肽对乳鼠心肌细胞肥大的抑制作用及可能机制_林杰.pdf

论著基础研究基金项目:福建省卫生健康科技计划项目省医学创新课题(2020CXB045)第一作者简介:林杰，本科，副主任医师，研究方向:心肌肥厚、心力衰竭和高血压。脑利钠肽对乳鼠心肌细胞肥大的抑制作用及可能机制林杰1罗瑞英1吴露楠2(宁德师范学院附属宁德市医院 1 心血管内科，2 病案室，福建省宁德市352199)【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(PPA)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 1 000 nmol/L 脑利钠肽+PE 组。除对照组外，其余组心肌细胞均给予 100 mol/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1 000 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 100 mol/L PE溶液处理，PE+抑制剂组心肌细胞在 PE 处理前先加入 1 mol/L GSK0660(PPA 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 1 mol/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 PC 检测各组心肌细胞的心房钠尿肽(ANP)和-肌球蛋白重链(-MHC)mNA 表达水平;采用 ELISA 测定 PPA 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 PPA 抑制剂实验中各组心肌细胞的 PPA、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-MHC mNA 表达水平升高，心肌细胞表面积增大(均 P 005);与 PE 组比较，各脑利钠肽干预组心肌细胞 ANP 和-MHC mNA 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、1 000 nmol/L 脑利钠肽+PE 组心肌细胞 ANP 和-MHC mNA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，1 000 nmol/L 脑利钠肽+PE 组心肌细胞 ANP 和-MHC mNA 表达水平降低(均 P 0 05)，而心肌细胞表面积差异无统计学意义(均 P 0 05)。(2)PE+抑制剂组、PE+脑利钠肽+抑制剂组、PE 组、正常组心肌细胞表面积依次减少(均 P 0 05);PE+抑制剂组、PE 组、PE+脑利钠肽+抑制剂组、正常组心肌细胞 ANP 和-MHC mNA 表达水平、FFA 含量依次降低，且 PPA、CPT-1 和 LCAD 蛋白表达水平依次升高(均 P 0 05)。结论脑利钠肽可抑制 PE 诱导的乳鼠心肌细胞肥大，其可能是通过上调 PPA 表达、促进脂肪酸氧化而发挥作用。【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;脂肪酸氧化;乳鼠;细胞实验【中图分类号】542 2【文献标识码】A【文章编号】0253-4304(2023)03-0296-06DOI:10 11675/j issn 0253-4304 2023 03 09Inhibitory effect and possible mechanism of brain natriuretic peptide oncardiomyocyte hypertrophy in suckling miceLIN Jie1，LUO uiying1，WU Lunan2(1 Department of Cardiovascular Medicine，2 oom of Medical ecords，Ningde MunicipalHospital of Ningde Normal University，Ningde 352199，Fujian，China)【Abstract】ObjectiveTo analyze the inhibitory effect of brain natriuretic peptide on cardiomyocyte hypertrophyin suckling mice induced by phenylephrine(PE)，and to explore its possible mechanism based on peroxisomeproliferator-activated receptor (PPA)and fatty acid oxidation Methods(1)Left ventricular tissues ofneonatal Kunming mice were obtained and prepared for cardiomyocytes(2)Cardiomyocytes were assigned to controlgroup，PE group，10 nmol/L brain natriuretic peptide+PE group，100 nmol/L brain natriuretic peptide+PE group，or692Guangxi Medical Journal，Feb 2023，Vol 45，No 31000 nmol/L brain natriuretic peptide+PE group Except for the control group，cardiomyocytes in the remaining groupsreceived treatment of 100 mol/L PE solution，and brain natriuretic peptide solution with corresponding concentrations wereadded to the 10 nmol/L brain natriuretic peptide+PE group，the 100 nmol/L brain natriuretic peptide+PE group，and the1000 nmol/L brain natriuretic peptide+PE group before PE treatment，respectively Cardiomyocytes of the control group werenormally cultured，without any medicine addition(3)Cardiomyocytes were randomly assigned to normal group，PEgroup，PE+inhibitor group，or PE+brain natriuretic peptide+inhibitor group Except for the normal group，cardiomyocytes in the remaining groups were treated with 100 mol/L PE solution，1 mol/L GSK0660(PPAinhibitor)solution was added to cardiomyocytes in the PE+inhibitor group before PE treatment，and 1 mol/LGSK0660 solution and 100 nmol/L brain natriuretic peptide solution were simultaneously added to cardiomyocytes inthe PE+brain natriuretic peptide+inhibitor group before PE treatment(4)After 48 hours of intervention，thesurface areas of cardiomyocytes were detected in various groups by the immunofluorescence technique The mNAexpressions of atrial natriuretic peptide(ANP)and beta myosin heavy chain(-MHC)were detected in cardiomyocytesin various groups by the real-time fluorescent quantitative PC The content of free fatty acid(FFA)of cardiomyocyteswas measured in various groups in the PPA inhibitor experiment by the ELISA The protein expressions of PPA，carnitine palmitoyltransferase 1(CPT-1)，and long-chain acyl-CoA dehydrogenase(LCAD)of cardiomyocytes in variousgroups in the PPA inhibitor experiment were detected by the Western blot esults(1)As compared with thecontrol group，the mNA expressions of ANP and-MHC of cardiomyocytes were elevated in the PE group，and thesurface area of cardiomyocytes was enlarged in the PE group(all P 0 05)Compared with the PE group，the mNAexpressions of ANP and-MHC of cardiomyocytes were declined in various brain natriuretic peptide intervention groups，andthe surface areas of cardiomyocytes were reduced in various brain natriuretic peptide intervention groups(all P 005)Compared with the 10 nmol/L brain natriuretic peptide+PE group，the mNA expressions of ANP and-MHC ofcardiomyocytes were declined，and the surface areas of cardiomyocytes were reduced in the 100 nmol/L brain natriureticpeptide+PE group and the 1000 nmol/L brain natriuretic peptide+PE group(all P 0 05)Compared with the100 nmol/L brain natriuretic peptid