真武汤对转基因扩张型心肌病小鼠心肌Acta1、Col3a1基因及蛋白表达的影响
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摘要
目的:本研究旨在观察转基因扩张型心肌病小鼠心肌组织Acta1、Col3a1基因及蛋白表达变化,探究真武汤防治扩张型心肌病的分子生物学机制。方法:将cT nT R141W基因转入C57BL/6J小鼠,建立了心肌组织特异表达cT nT R141W基因的转基因小鼠模型,以C57BL/6J小鼠15只作为空白对照组,模型组小鼠60只,随机分为模型对照组、西药组、中药高剂量组及中药中剂量组,中药组以真武汤高中剂量对模型小鼠治疗4周,卡托普利为阳性对照药物。4周后采用qRT-PCR法检测各组心肌组织Acta1、Col3a1相对表达量; Western Blot法检测各组心肌组织Acta1、Col3a1蛋白表达。结果:与空白对照组比较模型对照组Acta1、Col3a1基因及蛋白表达显著升高(P <0. 01),与模型对照组比较西药组及中药高中剂量组Acta1、Col3a1基因及蛋白表达均显著下降(P <0. 01)。结论:DCM的发病机制与心肌组织中Acta1与Col3a1mRNA及蛋白表达升高有关,真武汤通过下调心肌组织中Acta1与Col3a1 mRNA及蛋白表达,抑制心肌细胞肥大及纤维化过程,起到防治DCM的作用。
Objective: To observe the changes of Acta1 and Col3 a1 gene and protein expression in myocardium of mice with transgenic cardiomyopathy and explore the molecular mechanism of Zhenwu Decoction in the prevention and treatment of dilated cardiomyopathy. Methods: The cT nT R141 Wgene was transfected into C57 BL/6 J mice to establish a transgenic mouse model of cT-nT R141 Wgene specific for myocardium,fifteen C57 BL/6 J mice were used as blank control group. Sixty mice in the model group were randomly divided into model control group,western medicine group,high dose group and traditional Chinese medicine medium dose group. The rats in the traditional Chinese medicine group were treated with Zhenwu Decoction for 4 weeks and the captopril was the positive control drug. After 4 weeks,the expressions of Acta1 and Col3 a1 in myocardium were detected by qRT-PCR. The expressions of Acta1 and Col3 a1 protein were detected by Western Blot. Results: Compared with the blank control group,the expression of Acta1 and Col3 a1 gene and protein in the model control group were significantly increased( P < 0. 01).Compared with the model control group,the expressions of Acta1 and Col3 a1 gene and protein in the western medicine group and the high dose group were significantly decreased( P < 0. 01). Conclusion: The pathogenesis of DCM and the expressions of Acta1 and Col3 a1 mRNA and protein in myocardium are related to the expressions of Acta1 and Col3 a1 mRNA and protein in myocardium,and can inhibit the hypertrophy and fibrosis of cardiomyocytes.
Objective: To observe the changes of Acta1 and Col3 a1 gene and protein expression in myocardium of mice with transgenic cardiomyopathy and explore the molecular mechanism of Zhenwu Decoction in the prevention and treatment of dilated cardiomyopathy. Methods: The cT nT R141 Wgene was transfected into C57 BL/6 J mice to establish a transgenic mouse model of cT-nT R141 Wgene specific for myocardium,fifteen C57 BL/6 J mice were used as blank control group. Sixty mice in the model group were randomly divided into model control group,western medicine group,high dose group and traditional Chinese medicine medium dose group. The rats in the traditional Chinese medicine group were treated with Zhenwu Decoction for 4 weeks and the captopril was the positive control drug. After 4 weeks,the expressions of Acta1 and Col3 a1 in myocardium were detected by qRT-PCR. The expressions of Acta1 and Col3 a1 protein were detected by Western Blot. Results: Compared with the blank control group,the expression of Acta1 and Col3 a1 gene and protein in the model control group were significantly increased( P < 0. 01).Compared with the model control group,the expressions of Acta1 and Col3 a1 gene and protein in the western medicine group and the high dose group were significantly decreased( P < 0. 01). Conclusion: The pathogenesis of DCM and the expressions of Acta1 and Col3 a1 mRNA and protein in myocardium are related to the expressions of Acta1 and Col3 a1 mRNA and protein in myocardium,and can inhibit the hypertrophy and fibrosis of cardiomyocytes.
引文
[1]Pinto Y M,Elliott P M,Arbustini E,et al.Proposal for a revised definition of dilated cardiomyopathy,hypokinetic non-dilated cardiomyopathy,and its implications for clinical practice:a position statement of the ESC working group on myocardial and pericardial diseases[J].Eur Heart J,2016,37(23):1850-1858.
[2]Khalid S,Surr C,Neagu D,et al.Exploring the potential for secondary uses of Dementia Care Mapping(DCM)data for improving the quality of dementia care[J].Dementia(London),2017:40914331.
[3]Pfaffl M W.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucleic Acids Res,2001,29(9):e45.
[4]蔡婷.扩张型心肌病心力衰竭中医证型分布特征及其与NT-pro BNP、LVEF的相关性研究[D].武汉:湖北中医药大学,2013.
[5]鞠静,杜武勋.真武汤药效物质基础及温阳利水机制研究[J].吉林中医药,2016(7):719-723.
[6]白延平,刘智娜,高燕.强心宁煎剂联合曲美他嗪对慢性心衰左心室功能及血浆心钠素和内皮素水平的影响[J].四川中医,2016(9):69-71.
[7]吕洋,陈岩,杨积武.强心宁煎剂治疗扩张型心肌病临床观察[J].辽宁中医药大学学报,2015,42(6):201-203.
[8]Mohamed I A,El-Badri N,Zaher A.Dilated cardiomyopathy-induced disruption of basement membrane alters the lever systems acting on the heart[J].Med Hypotheses,2017,103:46-50.
[9]Mazurkiewicz L,Petryka J,Spiewak M,et al.Clinical and prognostic relevancy of left ventricular trabeculation assessed by cardiac magnetic resonance in patients with dilated cardiomyopathy[J].Kardiol Pol,2017.
[10]Wolff G,Lin Y,Karathanos A,et al.Implantable cardioverter/defibrillators for primary prevention in dilated cardiomyopathy postDANISH:an updated meta-analysis and systematic review of randomized controlled trials[J].Clin Res Cardiol,2017.
[11]Tanaka M,Hiroe M,Ito H,et al.Differential localization of atrial natriuretic peptide and skeletal alpha-actin messenger RNAs in left ventricular myocytes of patients with dilated cardiomyopathy[J].JAm Coll Cardiol,1995,26(1):85-92.
[12]Mirza M,Robinson P,Kremneva E,et al.The effect of mutations in alpha-tropomyosin(E40K and E54K)that cause familial dilated cardiomyopathy on the regulatory mechanism of cardiac muscle thin filaments[J].J Biol Chem,2007,282(18):13487-13497.
[13]Debold E P,Schmitt J P,Patlak J B,et al.Hypertrophic and dilated cardiomyopathy mutations differentially affect the molecular force generation of mouse alpha-cardiac myosin in the laser trap assay[J].Am J Physiol Heart Circ Physiol,2007,293(1):H284-H291.
[14]郝玉明,朱文玲.螺内酯对急性心肌梗死患者血浆Ⅲ型前胶原氨基末端肽含量的影响[J].临床心血管病杂志,2004,20(6):330-331.
[15]Weber K T.The lonely failing heart:a case for ECM genes[J].Cardiovasc Res,1995,30(6):835.
[16]王国钦,李瑞满,金伟,等.麦冬抑制大鼠心肌成纤维细胞胶原合成的作用及其机制[J].中国病理生理杂志,2013,29(4):615-618.
[2]Khalid S,Surr C,Neagu D,et al.Exploring the potential for secondary uses of Dementia Care Mapping(DCM)data for improving the quality of dementia care[J].Dementia(London),2017:40914331.
[3]Pfaffl M W.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucleic Acids Res,2001,29(9):e45.
[4]蔡婷.扩张型心肌病心力衰竭中医证型分布特征及其与NT-pro BNP、LVEF的相关性研究[D].武汉:湖北中医药大学,2013.
[5]鞠静,杜武勋.真武汤药效物质基础及温阳利水机制研究[J].吉林中医药,2016(7):719-723.
[6]白延平,刘智娜,高燕.强心宁煎剂联合曲美他嗪对慢性心衰左心室功能及血浆心钠素和内皮素水平的影响[J].四川中医,2016(9):69-71.
[7]吕洋,陈岩,杨积武.强心宁煎剂治疗扩张型心肌病临床观察[J].辽宁中医药大学学报,2015,42(6):201-203.
[8]Mohamed I A,El-Badri N,Zaher A.Dilated cardiomyopathy-induced disruption of basement membrane alters the lever systems acting on the heart[J].Med Hypotheses,2017,103:46-50.
[9]Mazurkiewicz L,Petryka J,Spiewak M,et al.Clinical and prognostic relevancy of left ventricular trabeculation assessed by cardiac magnetic resonance in patients with dilated cardiomyopathy[J].Kardiol Pol,2017.
[10]Wolff G,Lin Y,Karathanos A,et al.Implantable cardioverter/defibrillators for primary prevention in dilated cardiomyopathy postDANISH:an updated meta-analysis and systematic review of randomized controlled trials[J].Clin Res Cardiol,2017.
[11]Tanaka M,Hiroe M,Ito H,et al.Differential localization of atrial natriuretic peptide and skeletal alpha-actin messenger RNAs in left ventricular myocytes of patients with dilated cardiomyopathy[J].JAm Coll Cardiol,1995,26(1):85-92.
[12]Mirza M,Robinson P,Kremneva E,et al.The effect of mutations in alpha-tropomyosin(E40K and E54K)that cause familial dilated cardiomyopathy on the regulatory mechanism of cardiac muscle thin filaments[J].J Biol Chem,2007,282(18):13487-13497.
[13]Debold E P,Schmitt J P,Patlak J B,et al.Hypertrophic and dilated cardiomyopathy mutations differentially affect the molecular force generation of mouse alpha-cardiac myosin in the laser trap assay[J].Am J Physiol Heart Circ Physiol,2007,293(1):H284-H291.
[14]郝玉明,朱文玲.螺内酯对急性心肌梗死患者血浆Ⅲ型前胶原氨基末端肽含量的影响[J].临床心血管病杂志,2004,20(6):330-331.
[15]Weber K T.The lonely failing heart:a case for ECM genes[J].Cardiovasc Res,1995,30(6):835.
[16]王国钦,李瑞满,金伟,等.麦冬抑制大鼠心肌成纤维细胞胶原合成的作用及其机制[J].中国病理生理杂志,2013,29(4):615-618.