温阳通脉方通过JAK2/STAT3信号通路保护大鼠心肌缺血再灌注损伤
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摘要
目的探讨温阳通脉方是否通过Janus蛋白酪氨酸激酶2(JAK2)/信号转导和转录激活因子3(STAT3)信号转导通路,调节水通道蛋白(AQP)的表达,发挥对缺血再灌注心肌组织的保护作用。方法将30只雄性SD大鼠随机分为5组,每组6只:假手术组、模型组及温阳通脉方低剂量治疗组、中剂量治疗组和高剂量治疗组。将大鼠灌胃给药14天后,结扎心脏左冠状动脉前降支,按缺血30 min再灌注120 min的方法建立心肌缺血再灌注损伤模型。全自动生化分析仪检测各组大鼠的血清肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH);心电图检测大鼠ST段抬高情况;HE染色观察心脏组织病理形态学变化;免疫组织化学染色检测心肌组织中AQP1、AQP4蛋白的表达;Western blot检测JAK2、p-JAK2、STAT3、p-STAT3、AQP1、AQP4的蛋白表达。结果与假手术组比较,模型组大鼠血清中的CK-MB、LDH含量显著上升(P<0.01),缺血30 min与再灌注120 min后心电图的ST段显著抬高(P<0.01),HE染色显示模型组的心肌细胞损伤严重,AQP1、AQP4免疫组织化学表达明显增多(P<0.01),JAK2、p-JAK2、STAT3、p-STAT3、AQP1、AQP4的蛋白表达均明显升高(P<0.01)。与模型组相比,给药大鼠CK-MB、LDH水平明显降低(P<0.01),缺血30 min与再灌注120 min后的ST段均降低(P<0.01),心肌细胞损伤均可见不同程度减轻,AQP1、AQP4免疫组织化学表达明显减少,JAK2、p-JAK2、STAT3、p-STAT3整体的表达呈升高趋势,尤其是p-JAK2和p-STAT3的表达(P<0.01),AQP1、AQP4的水平均有不同程度降低(P<0.01)。结论温阳通脉方的心肌保护作用可能与激活JAK2/STAT3信号通路,下调AQP1和AQP4的表达有关。
Aim To explore whether Wenyang Tongmai decoction can regulate JAK2/STAT3 signal transduction pathway and the expression of aquaporins( AQP) to protect myocardial ischemia-reperfusion tissue. Methods Thirty male SD rats were randomly divided into 5 groups,6 in each group: sham operation group,ischemia-reperfusion group,low-dose treatment group,middle-dose treatment group,high-dose treatment group. After 14 days of intragastric administration,the model of myocardial ischemia-reperfusion injury was established by ligation of the left anterior descending coronary artery for 30 minutes and then releasing the ligated artery for 120 minutes. The serum levels of creatine kinase isoenzymes( CK-MB) and lactate dehydrogenase( LDH) were detected by automatic biochemical analyzer. The ST-segment elevation of the rats was counted in the electrocardiogram. Pathological changes in cardiac tissue were observed by HE staining. The expressions of AQP1 and AQP4 in myocardial tissue were detected by immunohistochemical staining.The protein expressions of JAK2,p-JAK2,STAT3,p-STAT3,AQP1 and AQP4 were detected by Western blot method.Results Compared with the sham group,the levels of CK-MB and LDH in the serum of the ischemia-reperfusion group were significantly increased( P<0. 01),and the ST-segments of the electrocardiogram were significantly elevated after ischemia for 30 minutes and reperfusion for 120 minutes( P<0. 01). HE staining showed that the myocardial cells in the ischemia-reperfusion group were severely damaged. The expressions of AQP1 and AQP4 in immunohistochemistry was significantly increased( P<0. 01). The results of Western blot showed that the expressions of JAK2,p-JAK2,STAT3,pSTAT3,AQP1 and AQP4 were significantly increased( P<0. 01). Compared with the ischemia-reperfusion group,the serum levels of CK-MB and LDH were significantly decreased in the treated rats( P<0. 01),and the ST-segments were decreased after 30 minutes ischemia and 120 minutes reperfusion( P<0. 01),myocardial cells' damage was alleviated to varying degrees in HE staining,the protein expressions of AQP1 and AQP4 in immunohistochemistry were significantly reduced,the expressions of JAK2,p-JAK2,STAT3 and p-STAT3 were generally increased,especially the expressions of pJAK2 and p-STAT3( P<0. 01),AQP1 and AQP4 expression levels were reduced to varying degrees( P<0. 01). Conclusion The cardioprotective effect of Wenyang Tongmai decoction may be related to activation of JAK2/STAT3 signaling pathway and down-regulation of AQP1 and AQP4 expression.
Aim To explore whether Wenyang Tongmai decoction can regulate JAK2/STAT3 signal transduction pathway and the expression of aquaporins( AQP) to protect myocardial ischemia-reperfusion tissue. Methods Thirty male SD rats were randomly divided into 5 groups,6 in each group: sham operation group,ischemia-reperfusion group,low-dose treatment group,middle-dose treatment group,high-dose treatment group. After 14 days of intragastric administration,the model of myocardial ischemia-reperfusion injury was established by ligation of the left anterior descending coronary artery for 30 minutes and then releasing the ligated artery for 120 minutes. The serum levels of creatine kinase isoenzymes( CK-MB) and lactate dehydrogenase( LDH) were detected by automatic biochemical analyzer. The ST-segment elevation of the rats was counted in the electrocardiogram. Pathological changes in cardiac tissue were observed by HE staining. The expressions of AQP1 and AQP4 in myocardial tissue were detected by immunohistochemical staining.The protein expressions of JAK2,p-JAK2,STAT3,p-STAT3,AQP1 and AQP4 were detected by Western blot method.Results Compared with the sham group,the levels of CK-MB and LDH in the serum of the ischemia-reperfusion group were significantly increased( P<0. 01),and the ST-segments of the electrocardiogram were significantly elevated after ischemia for 30 minutes and reperfusion for 120 minutes( P<0. 01). HE staining showed that the myocardial cells in the ischemia-reperfusion group were severely damaged. The expressions of AQP1 and AQP4 in immunohistochemistry was significantly increased( P<0. 01). The results of Western blot showed that the expressions of JAK2,p-JAK2,STAT3,pSTAT3,AQP1 and AQP4 were significantly increased( P<0. 01). Compared with the ischemia-reperfusion group,the serum levels of CK-MB and LDH were significantly decreased in the treated rats( P<0. 01),and the ST-segments were decreased after 30 minutes ischemia and 120 minutes reperfusion( P<0. 01),myocardial cells' damage was alleviated to varying degrees in HE staining,the protein expressions of AQP1 and AQP4 in immunohistochemistry were significantly reduced,the expressions of JAK2,p-JAK2,STAT3 and p-STAT3 were generally increased,especially the expressions of pJAK2 and p-STAT3( P<0. 01),AQP1 and AQP4 expression levels were reduced to varying degrees( P<0. 01). Conclusion The cardioprotective effect of Wenyang Tongmai decoction may be related to activation of JAK2/STAT3 signaling pathway and down-regulation of AQP1 and AQP4 expression.
引文
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[6]李楠楠,石月萍.加减枳实薤白桂枝汤通过AC/c AMP通路对缺血再灌注大鼠心肌组织水通道蛋白表达的影响[J].中国动脉硬化杂志,2018,26(3):237-244.
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[12]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.
[13]黄良珠,田英,龙石银.JAK/STAT信号通路及其负调控因子SOCS与动脉粥样硬化[J].中国动脉硬化杂志,2010,18(8):665-668.
[14]Zhang M,Wang X,Wang X,et al.Oxymatrine protects against myocardial injury via inhibition of JAK2/STAT3signal in rat septic shock[J].Mol Med Rep,2013,7(4):1293-1299.
[15]Yang FH,Xue L,Han ZQ,et al.Vaspin alleviates myocardial ischemia/reperfusion injury via activating autophagic flux and restoring lysosomal function[J].Biochem Biophys Res Commun,2018,503(2):501-507.
[16]Gross ER,Hsu AK,Gross GJ.The JAK/STAT pathway is essential for opioid-induced cardioprotection:JAK2 as a mediator of STAT3,Akt,and GSK-3 beta[J].Am JPhysiol Heart Circ Physiol,2006,291(2):H827-834.
[17]尹建国,张社兵,彭道泉.载脂蛋白AⅠ通过JAK2/STAT3信号通路抑制肿瘤坏死因子α的转录表达[J].中国动脉硬化杂志,2017,25(6):566-570.
[18]He Y,Li C,Ma Q,et al.Esculetin inhibits oxidative stress and apoptosis in H9c2 cardiomyocytes following hypoxia/reoxygenation injury[J].Biochem Biophys Res Commun,2018,501(1):139-144.
[19]Agre P.Aquaporin water channels:from atomic structure to clinical medicine[J].Nanomedicine,2006,2(4):266-267.
[20]Schorn C,Frey B,Lauber K,et al.Sodium overload and water influx activate the NALP3 inflammasome[J].J Biol Chem,2011,286(1):35-41.
[21]Kitchen P,Day RE,Salman MM,et al.Beyond water homeostasis:diverse functional roles of mammalian aquaporins[J].Biochim Biophys Acta,2015,1850(12):2410-2421.
[2]Turer AT,Hill JA.Pathogenesis of myocardial ischemiareperfusion injury and rationale for therapy[J].Am J Cardiol,2010,106(3):360-368.
[3]杨旭,邹澍宣.《金匮要略》胸痹治疗思想临床浅析[J].亚太传统医药,2017,13(11):75-76.
[4]曹凤华.枳实薤白桂枝汤预处理对大鼠心肌缺血再灌注损伤的保护作用[D].长春:长春中医药大学,2014:1-38.
[5]朱德建,陆曙.枳实薤白桂枝汤对不稳定型心绞痛患者Hcy及hs-CRP影响[J].辽宁中医药大学学报,2018,20(6):209-212.
[6]李楠楠,石月萍.加减枳实薤白桂枝汤通过AC/c AMP通路对缺血再灌注大鼠心肌组织水通道蛋白表达的影响[J].中国动脉硬化杂志,2018,26(3):237-244.
[7]刘春晓,石月萍.加减枳实薤白桂枝汤对大鼠心肌缺血再灌注损伤细胞凋亡及Caspase相关蛋白的影响[J].中药药理与临床,2017,33(1):13-16.
[8]李文珏.枳实薤白桂枝颗粒对心肌缺血作用及机制的初步探讨[D].哈尔滨:黑龙江中医药大学,2015:1-45.
[9]石月萍.加减枳实薤白桂枝汤治疗冠心病心绞痛[J].辽宁中医杂志,2009,36(6):962-963.
[10]石姗,孔祥英.枳实薤白桂枝汤治疗冠心病心绞痛的研究进展[J].世界最新医学信息文摘,2018,18(45):111-112.
[11]徐萍,石月萍.加减枳实薤白桂枝汤对大鼠心肌缺血再灌注过氧化损伤的影响[J].中药药理与临床,2017,33(3):16-20.
[12]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.
[13]黄良珠,田英,龙石银.JAK/STAT信号通路及其负调控因子SOCS与动脉粥样硬化[J].中国动脉硬化杂志,2010,18(8):665-668.
[14]Zhang M,Wang X,Wang X,et al.Oxymatrine protects against myocardial injury via inhibition of JAK2/STAT3signal in rat septic shock[J].Mol Med Rep,2013,7(4):1293-1299.
[15]Yang FH,Xue L,Han ZQ,et al.Vaspin alleviates myocardial ischemia/reperfusion injury via activating autophagic flux and restoring lysosomal function[J].Biochem Biophys Res Commun,2018,503(2):501-507.
[16]Gross ER,Hsu AK,Gross GJ.The JAK/STAT pathway is essential for opioid-induced cardioprotection:JAK2 as a mediator of STAT3,Akt,and GSK-3 beta[J].Am JPhysiol Heart Circ Physiol,2006,291(2):H827-834.
[17]尹建国,张社兵,彭道泉.载脂蛋白AⅠ通过JAK2/STAT3信号通路抑制肿瘤坏死因子α的转录表达[J].中国动脉硬化杂志,2017,25(6):566-570.
[18]He Y,Li C,Ma Q,et al.Esculetin inhibits oxidative stress and apoptosis in H9c2 cardiomyocytes following hypoxia/reoxygenation injury[J].Biochem Biophys Res Commun,2018,501(1):139-144.
[19]Agre P.Aquaporin water channels:from atomic structure to clinical medicine[J].Nanomedicine,2006,2(4):266-267.
[20]Schorn C,Frey B,Lauber K,et al.Sodium overload and water influx activate the NALP3 inflammasome[J].J Biol Chem,2011,286(1):35-41.
[21]Kitchen P,Day RE,Salman MM,et al.Beyond water homeostasis:diverse functional roles of mammalian aquaporins[J].Biochim Biophys Acta,2015,1850(12):2410-2421.