益母草碱对异丙肾上腺素诱导大鼠心肌重构的影响
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摘要
目的观察益母草碱对异丙肾上腺素(ISO)诱导大鼠心肌重构的影响,并探讨可能涉及的机制。方法 10只SD大鼠作正常对照组;另80只SD大鼠作为实验鼠,连续2周腹腔注射ISO诱导心肌重构动物模型。造模成功后将实验鼠随机分为5组:模型组,益母草碱低(7.5mg/(kg·d))、中(15mg/(kg·d))、高(30mg/(kg·d))剂量组以及p38丝裂原活化蛋白激酶(p38MAPK)抑制剂组(0.3mg/(kg·d)),予相应药物腹腔注射2周后,HE染色观察左心室肌组织形态学变化;免疫组织化学法观察左心室肌组织转化生长因子-β1(TGF-β1)的表达;ELISA法及硝酸还原酶法检测血清内皮素(ET-1)和一氧化氮(NO)含量;qPCR法检测左心室肌组织p38MAPK、肌细胞增强因子2(MEF2)、β?肌球蛋白重链(β?MHC)、α?肌球蛋白重链(α?MHC)mRNA的表达水平;Western blot法检测左心室肌组织ET-1、磷酸化p38MAPK(p-p38MAPK)和MEF2蛋白的表达水平。结果与模型组比较,高剂量益母草碱能明显改善大鼠心肌重构病理改变,减少TGF-β1的表达(P<0.05);减少血清ET-1含量,升高NO含量(P<0.05);下调p38MAPK、MEF2和β?MHC mRNA的表达水平及ET-1、p-p38MAPK和MEF2蛋白的表达水平,上调α?MHC mRNA的表达水平(P<0.05)。结论益母草碱有抑制ISO诱导的大鼠心肌重构的作用,其机制可能与抑制p38MAPK/MEF2信号通路有关。
OBJECTIVE To observe the effect of leonurine on rats with ventricular remodeling induced by isoproterenol(ISO)and to explore the possible mechanism involved.METHODS SD rats(n=10)were used as normal control group,and80 rats were given ISO by intraperitoneal injection daily for 2 weeks to establish the model of ventricular remodeling.The model rats were divided into 5 groups randomly as model group,low-dose leonurine(7.5 mg/(kg·d))group,middle-dose leonurine(15 mg/(kg·d))group,high-dose leonurine(30 mg/(kg·d))group and p38 MAPK inhibitor(0.3 mg/(kg·d))group.After the treatment for 2 weeks,the pathological change of left ventricular myocardial tissues was observed by HE staining,and the expression of TGF-β1 was determined by the method of immunohistochemistry.The serum concentrations of ET-1 and NO were measured by ELISA and nitrate reductase methods,respectively.The expression of p38 MAPK,MEF2,β?MHC andα?MHC mRNA was detected by qPCR,and the protein expression of ET-1,p-p38 MAPK and MEF2 was determined by Western blot.RESULTS Compared with model group,the pathological change of ventricular remodeling in high-dose leonurine group was attenuated,and the serum concentrations of NO and the mRNA expression ofα?MHC in left ventricular myocardial tissues of high-dose leonurine group were higher(P<0.05).The expression of TGF-β1,the serum concentrations of ET-1,the mRNA expression of p38 MAPK,MEF2 andβ?MHC mRNA,and the protein expression of ET-1,p-p38 MAPK and MEF2 were lower than those in model group(P<0.05).CONCLUSION Leonurine attenuates ventricular remodeling in the rats induced by ISO,and it is potentially associated with inhibiting p38 MAPK/MEF2 signaling pathway.
OBJECTIVE To observe the effect of leonurine on rats with ventricular remodeling induced by isoproterenol(ISO)and to explore the possible mechanism involved.METHODS SD rats(n=10)were used as normal control group,and80 rats were given ISO by intraperitoneal injection daily for 2 weeks to establish the model of ventricular remodeling.The model rats were divided into 5 groups randomly as model group,low-dose leonurine(7.5 mg/(kg·d))group,middle-dose leonurine(15 mg/(kg·d))group,high-dose leonurine(30 mg/(kg·d))group and p38 MAPK inhibitor(0.3 mg/(kg·d))group.After the treatment for 2 weeks,the pathological change of left ventricular myocardial tissues was observed by HE staining,and the expression of TGF-β1 was determined by the method of immunohistochemistry.The serum concentrations of ET-1 and NO were measured by ELISA and nitrate reductase methods,respectively.The expression of p38 MAPK,MEF2,β?MHC andα?MHC mRNA was detected by qPCR,and the protein expression of ET-1,p-p38 MAPK and MEF2 was determined by Western blot.RESULTS Compared with model group,the pathological change of ventricular remodeling in high-dose leonurine group was attenuated,and the serum concentrations of NO and the mRNA expression ofα?MHC in left ventricular myocardial tissues of high-dose leonurine group were higher(P<0.05).The expression of TGF-β1,the serum concentrations of ET-1,the mRNA expression of p38 MAPK,MEF2 andβ?MHC mRNA,and the protein expression of ET-1,p-p38 MAPK and MEF2 were lower than those in model group(P<0.05).CONCLUSION Leonurine attenuates ventricular remodeling in the rats induced by ISO,and it is potentially associated with inhibiting p38 MAPK/MEF2 signaling pathway.
引文
[1]AZEVEDO PS,POLEGATO BF,MINICUCCI MF,et al.Cardiac remodeling:Concepts,clinical impact,pathophysiological mechanisms and pharmacologic treatment[J].Arq Bras Cardiol,2016,106(1):62-69.
[2]ZHU YZ,WU W,ZHU Q,et al.Discovery of Leonuri and therapeutical applications:From bench to bedside[J].Pharmacol Ther,2018,188:26-35.
[3]梁赵文,罗建华,杨冬花,等.益母草碱对慢性心力衰竭心肌重构大鼠心房利钠肽、血管紧张素Ⅱ影响的研究[J].贵州医药,2016,40(12):1235-1238.
[4]NGUYEN MN,KIRIAZIS H,GAO XM,et al.Cardiac fibrosis and arrhythmo-genesis[J].Compr Physiol,2017,7(3):1009-1049.
[5]张勇涛,蒋凡.生理性和病理性心肌肥厚的信号传导机制[J].中华心血管病杂志,2015,43(3):277-280.
[6]MA Y,ZOU H,ZHU XX,et al.Transforming growth factorβ:A potential biomarker and therapeutic target of ventricular remodeling[J].Oncotarget,2017,8(32):53780-53790.
[7]KONG P,CHRISTIA P,FRANGOGIANNIS NG.The pathogenesis of cardiac fibrosis[J].Cell Mol Life Sci,2014,71(4):549-574.
[8]宋冰,丁利.硫化氢抑制p38MAPK和NF-κB p65信号通路活性改善糖尿病心肌纤维化的作用和机制[J].解放军医学杂志,2016,41(11):902-907.
[9]ARABACILAR P,MARBER M.The case for inhibiting p38mitogen-activated protein kinase in heart failure[J].Front Pharmacol,2015,6:102.
[10]DAVENPORT AP,HYNDMAN KA,DHAUN N,et al.Endothelin[J].Pharmacol Rev,2016,68(2):357-418.
[11]JEN HL,LIU PL,CHEN YH,et al.Peroxisome proliferator-activated receptorαreduces endothelin-1-caused cardiomyocyte hypertrophy by inhibiting nuclear factor-κB and adiponectin[J].Mediators Inflamm,2016,2016:5609121.
[12]WENG X,YU L,LIANG P,et al.A crosstalk between chromatin remodeling and histone H3K4methyltransferase complexes in endothelial cells regulates angiotensinⅡ?induced cardiac hypertrophy[J].J Mol Cell Cardiol,2015,82:48-58.
[13]GONG X,MING X,DENG P,et al.Mechanisms regulating the nuclear translocation of p38 MAP kinase[J].J Cell Biochem,2010,110(6):1420-1429.
[14]CHEN X,GAO B,PONNUSAMY M,et al.MEF2signaling and human diseases[J].Oncotarget,2017,8(67):112152-112165.
[15]ZHAO M,NEW L,KRAVCHENKO VV,et al.Regulation of the MEF2family of transcription factors by p38[J].Mol Cell Biol,1999,19(1):21-30.
[16]HAN J,JIANG Y,LI Z,et al.Activation of the transcription factor MEF2Cby the MAP kinase p38in inflammation[J].Nature,1997,386(6622):296-299.
[17]DADSON K,TURDI S,HASHEMI S,et al.Adiponectin is required for cardiac MEF2activation during pressure overload induced hypertrophy[J].J Mol Cell Cardiol,2015,86:102-109.
[2]ZHU YZ,WU W,ZHU Q,et al.Discovery of Leonuri and therapeutical applications:From bench to bedside[J].Pharmacol Ther,2018,188:26-35.
[3]梁赵文,罗建华,杨冬花,等.益母草碱对慢性心力衰竭心肌重构大鼠心房利钠肽、血管紧张素Ⅱ影响的研究[J].贵州医药,2016,40(12):1235-1238.
[4]NGUYEN MN,KIRIAZIS H,GAO XM,et al.Cardiac fibrosis and arrhythmo-genesis[J].Compr Physiol,2017,7(3):1009-1049.
[5]张勇涛,蒋凡.生理性和病理性心肌肥厚的信号传导机制[J].中华心血管病杂志,2015,43(3):277-280.
[6]MA Y,ZOU H,ZHU XX,et al.Transforming growth factorβ:A potential biomarker and therapeutic target of ventricular remodeling[J].Oncotarget,2017,8(32):53780-53790.
[7]KONG P,CHRISTIA P,FRANGOGIANNIS NG.The pathogenesis of cardiac fibrosis[J].Cell Mol Life Sci,2014,71(4):549-574.
[8]宋冰,丁利.硫化氢抑制p38MAPK和NF-κB p65信号通路活性改善糖尿病心肌纤维化的作用和机制[J].解放军医学杂志,2016,41(11):902-907.
[9]ARABACILAR P,MARBER M.The case for inhibiting p38mitogen-activated protein kinase in heart failure[J].Front Pharmacol,2015,6:102.
[10]DAVENPORT AP,HYNDMAN KA,DHAUN N,et al.Endothelin[J].Pharmacol Rev,2016,68(2):357-418.
[11]JEN HL,LIU PL,CHEN YH,et al.Peroxisome proliferator-activated receptorαreduces endothelin-1-caused cardiomyocyte hypertrophy by inhibiting nuclear factor-κB and adiponectin[J].Mediators Inflamm,2016,2016:5609121.
[12]WENG X,YU L,LIANG P,et al.A crosstalk between chromatin remodeling and histone H3K4methyltransferase complexes in endothelial cells regulates angiotensinⅡ?induced cardiac hypertrophy[J].J Mol Cell Cardiol,2015,82:48-58.
[13]GONG X,MING X,DENG P,et al.Mechanisms regulating the nuclear translocation of p38 MAP kinase[J].J Cell Biochem,2010,110(6):1420-1429.
[14]CHEN X,GAO B,PONNUSAMY M,et al.MEF2signaling and human diseases[J].Oncotarget,2017,8(67):112152-112165.
[15]ZHAO M,NEW L,KRAVCHENKO VV,et al.Regulation of the MEF2family of transcription factors by p38[J].Mol Cell Biol,1999,19(1):21-30.
[16]HAN J,JIANG Y,LI Z,et al.Activation of the transcription factor MEF2Cby the MAP kinase p38in inflammation[J].Nature,1997,386(6622):296-299.
[17]DADSON K,TURDI S,HASHEMI S,et al.Adiponectin is required for cardiac MEF2activation during pressure overload induced hypertrophy[J].J Mol Cell Cardiol,2015,86:102-109.