丹红注射液对心梗大鼠心肌细胞凋亡及JAK2/STAT3信号通路的影响
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摘要
目的:研究中药复方制剂丹红注射液对冠脉结扎大鼠心肌细胞凋亡的影响及相关分子机制。方法:取40只雄性Sprague-Dawley(SD)大鼠随机分为假手术组、模型组、丹红注射液低、中、高剂量组,左前降支冠状动脉结扎法造成急性心梗模型,采用超声心动图法记录大鼠心功能,TUNEL标记法检测心肌组织细胞凋亡状况,蛋白免疫印迹法检测心肌组织B细胞淋巴瘤/白血病-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)的表达、天冬氨酸蛋白水解酶-3(Caspase-3)的活化、Janus激酶2(Janus kinase 2,JAK2)和信号转导及转录激活因子3(signal transducer and activator of transcription 3, STAT3)的磷酸化水平。结果:丹红注射液能剂量依赖性地升高大鼠心功能指标左室射血分数(left ventricular ejection fraction, LVEF)和左室短轴缩短率(left ventricular fractional shortening, LVFS)的值,减少缺血损伤诱导的心肌细胞凋亡,增加抗凋亡基因Bcl-2的表达,减少促凋亡基因Bax的表达及Caspase-3的活化,并上调JAK2和STAT3的磷酸化水平。结论:丹红注射液可显著改善大鼠心功能,抑制缺血诱导的心肌细胞凋亡,其机制可能与调节凋亡相关基因及JAK2/STAT3信号通路参与调控有关。
Objective: To investigate the effects ofDanhong injection(DHI) on cardiomyocyte apoptosisand related molecular mechanismsin rats with myocardial infarction(MI). Methods: 40 male Sprague-Dawleyrats were randomly divided into five groups:Sham-operated group; MI+ saline group; MI+ DHI 0.5 m L/kg group; MI+ DHI 1 mL/kg group; MI+ DHI 2 mL/kg group, and acuteMIwere induced by ligation of left anteriordescending coronary artery. After intravenous injection for 7 days, the cardiac function of ratswasassessed by echocardiography. Then the cardiomyocyte apoptosis index was detected by terminal deoxynucleotidyl transferase d UTP nick end labeling(TUNEL) technology, and the expression of B-cell lymphoma/lewkmia-2(Bcl-2), Bcl-2 associated X protein(Bax), Cleaved cysteine aspartate protease-3(Caspase-3), and phosphorylation levels of Januskinase 2(JAK2) and signal transducerand activator of transcription 3(STAT3)inmyocardiumwere determined by western blot analysis. Results: DHI restored cardiac function evidenced by increases of left ventricular ejection fraction(LVEF) and left ventricular fractional shortening(LVFS)in a dose-dependent manner, and inhibited cardiomyocytes apoptosis induced by ischemia injury. Furthermore, DHI increased Bcl-2 expression, while decreased Bax and active caspase-3 expression. The phosphorylation of JAK2/STAT3 was also upregulated by DHI. Conclusion: DHI couldsignificantly improve left ventricular function of MI rats, and inhibit ischemia-induced cardiomyocytes apoptosis. These beneficial effects may be associated with the regulation of apoptosis related gene and JAK2/STAT3 signal pathway.
Objective: To investigate the effects ofDanhong injection(DHI) on cardiomyocyte apoptosisand related molecular mechanismsin rats with myocardial infarction(MI). Methods: 40 male Sprague-Dawleyrats were randomly divided into five groups:Sham-operated group; MI+ saline group; MI+ DHI 0.5 m L/kg group; MI+ DHI 1 mL/kg group; MI+ DHI 2 mL/kg group, and acuteMIwere induced by ligation of left anteriordescending coronary artery. After intravenous injection for 7 days, the cardiac function of ratswasassessed by echocardiography. Then the cardiomyocyte apoptosis index was detected by terminal deoxynucleotidyl transferase d UTP nick end labeling(TUNEL) technology, and the expression of B-cell lymphoma/lewkmia-2(Bcl-2), Bcl-2 associated X protein(Bax), Cleaved cysteine aspartate protease-3(Caspase-3), and phosphorylation levels of Januskinase 2(JAK2) and signal transducerand activator of transcription 3(STAT3)inmyocardiumwere determined by western blot analysis. Results: DHI restored cardiac function evidenced by increases of left ventricular ejection fraction(LVEF) and left ventricular fractional shortening(LVFS)in a dose-dependent manner, and inhibited cardiomyocytes apoptosis induced by ischemia injury. Furthermore, DHI increased Bcl-2 expression, while decreased Bax and active caspase-3 expression. The phosphorylation of JAK2/STAT3 was also upregulated by DHI. Conclusion: DHI couldsignificantly improve left ventricular function of MI rats, and inhibit ischemia-induced cardiomyocytes apoptosis. These beneficial effects may be associated with the regulation of apoptosis related gene and JAK2/STAT3 signal pathway.
引文
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[12]王军,张虹,谢凤杰,等.甘草酸预处理对大鼠心肌缺血再灌注损伤的保护作用及Bcl-2和Bax表达的影响[J].中国实验方剂学杂志,2018,24(6):126-132
[13]Eichhorn JM,Sakurikar N,Alford SE,et al.Critical role of antiapoptotic Bcl-2 protein phosphorylation in mitoticdeath[J].Cell Death Dis,2013,4:e834
[14]Julien O,Wells JA.Caspases and their substrates[J].Cell Death Differ,2017,24(8):1380-1389
[15]苏宁,卢芬萍,朱明明,等.黄芪甲苷联合羟基红花黄色素A对小鼠心肌梗死的保护作用[J].中国实验方剂学杂志,2018,24(2):98-103
[16]王筱冰,张小翠,夏妙红,等.Caspase的活化机制[J].现代生物医学进展,2006,6(3):53-55
[17]Villarino AV,Kanno Y,O'Shea JJ.Mechanisms and consequences of Jak-STAT signaling in the immune system[J].Nat Immunol,2017,18(4):374-384
[18]Oshima Y,Fujio Y,Nakanishi T,et al.STAT3 mediates cardioprotection against ischemia/reperfusion injury through metallothionein induction in the heart[J].Cardiovasc Res,2005,65(2):428-435
[19]刘永强,冷玉芳,叶元梅,等.JAK2/STAT3信号通路在羟考酮预处理预防心肌缺血再灌注损伤中的作用[J].兰州大学学报(医学版),2017,43(1):26-30
[20]Hattori R,Maulik N,Otani H,et al.Role of STAT3 in ischemic preconditioning[J].J Mol Cell Cardiol,2001,33(11):1929-1936
[21]Boengler K,Hilfiker-Kleiner D,Heusch G,et al.Inhibition of permeability transition pore opening bymitochondrial STAT3 and its role in myocardial ischemia/reperfusion[J].Basic Res Cardiol,2010,105(6):771-785
[2]Guan Y,Yin Y,Zhu YR,et al.Dissection of mechanisms of a Chinese medicinal formula:Danhong injection therapy for myocardial ischemia/reperfusion injury in vivo and in vitro[J].Evid Based Complement Alternat Med,2013,2013(972370):1-12
[3]殷英,卫国,王艳华,等.丹红注射液促进心梗模型大鼠缺血心肌血管新生[J].中成药,2016,38(9):1893-1897
[4]Kisseleva T,Bhattacharya S,Braunstein J,et al.Signaling through the JAK/STAT pathway,recent advances and future challenges[J].Gene,2002,285(1-2):1-24
[5]冷雪,宋囡,臧安缘,等.JAK2/STAT3信号通路在人参皂苷Re预处理预防异丙肾上腺素致急性心肌缺血损伤中的作用[J].中国药理学通报,2018,34(1):103-107
[6]Hojo Y,Saito T,Kondo H.Role of apoptosis in left ventricular remodeling after acute myocardial infarction[J].J Cardiol,2012,60(2):91-92
[7]Tsoporis JT,Izhar S,Desjardins JF,et al.Conditional cardiac overexpression of S100A6 attenuates myocyte hypertrophy and apoptosis following myocardial infarction[J].Curr Pharm Des,2014,20(12):1941-1949
[8]Zhang X,Wang H,Chang Y,et al.An overview of meta-Analyses of danhong injection for unstable angina[J].Evid Based Complement Alternat Med,2015,2015(358028):1-8
[9]Duan ZZ,Li YH,Li YY,et al.Danhong injection protects cardiomyocytes againsthypoxia/reoxygenation-and H2O2-induced injury by inhibitingmitochondrial permeability transition pore opening[J].JEthnopharmacol,2015,175:617-25
[10]卫国,殷英,段佳林,等.丹红注射液中5种主要活性成分促血管新生作用研究[J].现代生物医学进展,2017,17(17):3230-3233+3249
[11]Reed JC.Proapoptotic multidomain Bcl-2/Bax-family proteins:mechanisms,physiological roles,and therapeutic opportunities[J].Cell Death Differ,2006,13(8):1378-1386
[12]王军,张虹,谢凤杰,等.甘草酸预处理对大鼠心肌缺血再灌注损伤的保护作用及Bcl-2和Bax表达的影响[J].中国实验方剂学杂志,2018,24(6):126-132
[13]Eichhorn JM,Sakurikar N,Alford SE,et al.Critical role of antiapoptotic Bcl-2 protein phosphorylation in mitoticdeath[J].Cell Death Dis,2013,4:e834
[14]Julien O,Wells JA.Caspases and their substrates[J].Cell Death Differ,2017,24(8):1380-1389
[15]苏宁,卢芬萍,朱明明,等.黄芪甲苷联合羟基红花黄色素A对小鼠心肌梗死的保护作用[J].中国实验方剂学杂志,2018,24(2):98-103
[16]王筱冰,张小翠,夏妙红,等.Caspase的活化机制[J].现代生物医学进展,2006,6(3):53-55
[17]Villarino AV,Kanno Y,O'Shea JJ.Mechanisms and consequences of Jak-STAT signaling in the immune system[J].Nat Immunol,2017,18(4):374-384
[18]Oshima Y,Fujio Y,Nakanishi T,et al.STAT3 mediates cardioprotection against ischemia/reperfusion injury through metallothionein induction in the heart[J].Cardiovasc Res,2005,65(2):428-435
[19]刘永强,冷玉芳,叶元梅,等.JAK2/STAT3信号通路在羟考酮预处理预防心肌缺血再灌注损伤中的作用[J].兰州大学学报(医学版),2017,43(1):26-30
[20]Hattori R,Maulik N,Otani H,et al.Role of STAT3 in ischemic preconditioning[J].J Mol Cell Cardiol,2001,33(11):1929-1936
[21]Boengler K,Hilfiker-Kleiner D,Heusch G,et al.Inhibition of permeability transition pore opening bymitochondrial STAT3 and its role in myocardial ischemia/reperfusion[J].Basic Res Cardiol,2010,105(6):771-785