复方丹参片调控AT_1介导花生四烯酸PLA2-COX2/5-LOX代谢途径防治心力衰竭的机制研究
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摘要
心肌纤维化(myocardial fibrosis,MF)是心肌梗死(myocardial infarction,MI)向心力衰竭(heart failure,HF)转化的重要病理生理机制,而心肌细胞内花生四烯酸(arachidonic acid,AA)代谢紊乱在MF的发生发展过程中发挥着重要作用。有文献报道复方丹参片具有抗纤维化的作用,但其具体作用机制尚不明确。该实验通过左冠状动脉前降支结扎术制备MI后HF动物模型,利用超声、血流动力学、病理切片染色、TUNEL和Western blot等技术进行检测,探究复方丹参片抑制MF进而防治HF的药效及其作用机制。结果显示,复方丹参片可以显著提高MI大鼠的左室短轴缩短率(fractional shortening,FS)、射血分数(ejection fraction,EF)、左心室收缩压(left ventricular systolic pressure,LVSP)等心功能指标;此外,可以显著降低血清肌酸激酶同工酶(creatine kinase-MB,CK-MB)、乳酸脱氢酶(lactate dehydrogenase,LDH)的含量;病理结果显示,复方丹参片可以有效减少缺血区心肌组织的炎性细胞浸润,抑制纤维化程度与心肌细胞凋亡;Western blot结果显示复方丹参片可以下调缺血心肌组织中AT1,MMP2,MMP9的含量,并且上调AT2的含量;此外,复方丹参片还可以抑制缺血心肌组织中AA代谢通路中PLA2,P450,COX2,5-LOX的表达。因此,复方丹参片能有效抑制大鼠心肌梗死后缺血区域的纤维化程度,进而延缓其HF进程;其机制与复方丹参片调节AT1介导的AA相关的PLA2-COX2代谢途径相关。
Myocardial fibrosis( MF) is an important pathological change involved in the progress from myocardial infarction( MI)to heart failure( HF). Metabolic disorder of arachidonic acid( AA) in cardiomyocytes plays an important role in process of MF. Fufang Danshen tablets is a traditional Chinese medicine( TCM),which showed significant effect on coronary heart diseases and anti-MF.However,the underlying mechanism of anti-MF remains unclear. In this study,HF animal model of myocardial infarction was established by ligation of left anterior descending coronary artery. The heart function of rats in each group was evaluated by echocardiography and hemodynamic measurement. Histological examination,TUNEL and Western blot were used to detect the levels of MF and proteins related to AA metabolism. As a result,MI significantly decreased the levels of ejection fraction( EF),ejection fraction( FS) and left ventricular systolic pressure( LVSP),and these decreases were significantly improved by the treatment of Fufang Danshen tablets. Besides,Fufang Danshen tablets treatment down-regulated the levels of creatine kinase-MB( CK-MB) and lactate dehydrogenase( LDH)in serum. HE,Masson and TUNEL staining results showed that Fufang Danshen tablets treatment could inhibit the inflammatory cells infiltration and attenuate the fibrosis and apoptosis to exert cardioprotective effect. Western blot indicated that Fufang Danshen tablets treatment down-regulated the expressions of AT1,MMP2,MM9,while up-regulated the expression of AT2 to inhibit MF. Further mechanism study indicated that Fufang Danshen tablets inhibited MF by down-regulated the expressions of AA metabolism,such as PLA2,P450,COX2 and 5-LOX. In summary,Fufang Danshen tablets can effectively inhibit MF in the ischemic area after MI in rats.The mechanism is related to the regulation of AT1-mediated PLA2-COX2 metabolic pathway.
Myocardial fibrosis( MF) is an important pathological change involved in the progress from myocardial infarction( MI)to heart failure( HF). Metabolic disorder of arachidonic acid( AA) in cardiomyocytes plays an important role in process of MF. Fufang Danshen tablets is a traditional Chinese medicine( TCM),which showed significant effect on coronary heart diseases and anti-MF.However,the underlying mechanism of anti-MF remains unclear. In this study,HF animal model of myocardial infarction was established by ligation of left anterior descending coronary artery. The heart function of rats in each group was evaluated by echocardiography and hemodynamic measurement. Histological examination,TUNEL and Western blot were used to detect the levels of MF and proteins related to AA metabolism. As a result,MI significantly decreased the levels of ejection fraction( EF),ejection fraction( FS) and left ventricular systolic pressure( LVSP),and these decreases were significantly improved by the treatment of Fufang Danshen tablets. Besides,Fufang Danshen tablets treatment down-regulated the levels of creatine kinase-MB( CK-MB) and lactate dehydrogenase( LDH)in serum. HE,Masson and TUNEL staining results showed that Fufang Danshen tablets treatment could inhibit the inflammatory cells infiltration and attenuate the fibrosis and apoptosis to exert cardioprotective effect. Western blot indicated that Fufang Danshen tablets treatment down-regulated the expressions of AT1,MMP2,MM9,while up-regulated the expression of AT2 to inhibit MF. Further mechanism study indicated that Fufang Danshen tablets inhibited MF by down-regulated the expressions of AA metabolism,such as PLA2,P450,COX2 and 5-LOX. In summary,Fufang Danshen tablets can effectively inhibit MF in the ischemic area after MI in rats.The mechanism is related to the regulation of AT1-mediated PLA2-COX2 metabolic pathway.
引文
[1]O'Donoghue M L,Morrow D A,Cannon C P,et al.Multimarker risk stratification in patients with acute myocardial infarction[J].J Am Heart Assoc,2016,5(5):e002586.
[2]Talman V,Ruskoaho H.Cardiac fibrosis in myocardial infarction—from repair and remodeling to regeneration[J].Cell Tissue Res,2016,365(3):563.
[3]Morrone D,Marzilli M.Role of RAAS inhibition in preventing left ventricular remodeling in patients post myocardial infarction[J].Heart Metab,2010(47):9.
[4]Roth L,Schrijvers D M,Martinet W,et al.AngiotensinⅡincreases coronary fibrosis,cardiac hypertrophy and the incidence of myocardial infarctions in Apo E-/-Fbn1C1039G+/-mice[J].Acta Cardiol,2016,71(4):483.
[5]Namsolleck P,Recarti C,Foulquier S,et al.AT(2)receptor and tissue injury:therapeutic implications.[J].Curr Hypertens Rep,2014,16(2):416.
[6]Balsinde J,Winstead M V,Dennis E A.Phospholipase A2,regulation of arachidonic acid mobilization[J].Febs Lett,2002,531(1):2.
[7]Wang J,Lu L,Wang Y,et al.Qishenyiqi dropping pill attenuates myocardial fibrosis in rats by inhibiting RAAS-mediated arachidonic acid inflammation[J].J Ethnopharmacol,2015,176(16):375.
[8]Li C,Wang J,Wang Q,et al.Qishen granules inhibit myocardial inflammation injury through regulating arachidonic acid metabolism[J].Sci Rep,2016,6:36949.
[9]Hsu C C,Lai S C.Matrix metalloproteinase-2,-9 and-13 are involved in fibronectin degradation of rat lung granulomatous fibrosis caused by Angiostrongylus cantonensis[J].Int J Exp Pathol,2007,88(6):437.
[10]Ito H,Duxbury M,Benoit E,et al.Fibronectin-induced COX-2mediates MMP-2 expression and invasiveness of rhabdomyosarcoma[J].Biochem Biophys Res Commun,2004,318(2):594.
[11]叶华.用代谢组学方法评价复方丹参片对血瘀动物的作用[D].西安:陕西师范大学,2011.
[12]杨暋慧,张暋浩,朱尔佳,等.复方丹参膜控缓释片与复方丹参片在Beagle犬体内的药动学及生物利用度的比较研究[J].今日药学,2014,24(6):393.
[13]潘桂湘.复方丹参方主要化学成分的研究[D].北京:北京中医药大学,2002.
[14]成娟,蔡文辉,朱云杰.肝苏颗粒联合复方丹参片治疗肝纤维化28例[J].中西医结合肝病杂志,2001,11(5):301.
[15]郭玉东,李丽,左泽平,等.复方丹参片对大鼠急性心肌缺血的保护作用[J].解放军药学学报,2016(6):497.
[16]Wang Y,Li C,Ouyang Y,et al.Cardioprotective effects of Qishenyiqi mediated by angiotensinⅡtype 1 receptor blockade and enhancing angiotensin-converting enzyme 2[J].Evid-based Complement Altern Med,2012,2012(6):978127.
[17]王蕾,赵明镜,杨涛,等.从心电图和超声心动图相关性分析研究心肌梗死后心衰模型的早期评价和筛选方法[J].中西医结合心脑血管病杂志,2017(22):2816.
[18]Amani M,Jeddi S,Ahmadiasl N,et al.Effect of HEMADO on level of CK-MB and LDH enzymes after ischemia/reperfusion injury in isolated rat heart[J].Bio Impacts,2013,3(2):101.
[19]De Mello W C.Intracellular angiotensinⅡdisrupts chemical communication and impairs metabolic cooperation between cardiac myocytes[J].Peptides,2015,72:57.
[20]Higuchi S,Ohtsu H,Suzuki H,et al.AngiotensinⅡsignal transduction through the AT1 receptor:novel insights into mechanisms and pathophysiology[J].Clin Sci,2007,112(8):417.
[21]Jenkins C M,Cedars A,Gross R W.Eicosanoid signalling pathways in the heart[J].Cardiovasc Res,2009,82(2):240.
[22]López B,González A,Querejeta R,et al.Myocardial COX-2overexpression and fibrosis in hypertensive heart disease.Role of angiotensinⅡ[J].Amer J Hypertens,2005,18(5):A233.
[23]Callejas N A,Casado M,Díazguerra M J,et al.Expression of cyclooxygenase-2 promotes the release of matrix metalloproteinase-2 and-9 in fetal rat hepatocytes[J].Hepatology,2001,33(4):860.
[24]Rhinehart K,Handelsman C A,Silldorff E P,et al.ANGⅡAT2 receptor modulates AT1 receptor-mediated descending vasa recta endothelial Ca2+signaling[J].Amer J Physiol Heart Circ Phy,2003,284(3):H779.
[25]马艳秋.丹参酮ⅡA对大鼠心脏纤维化作用及机制的研究[D].郑州:郑州大学,2016.
[26]张翼宙,卢冬冬,董颖,等.丹参酮ⅡA对大鼠肝纤维化的干预作用及其调控AngⅡ的分子机制[J].浙江中医药大学学报,2017,41(1):1.
[27]郑娴.人参皂苷Rb1对心衰大鼠心肌重构的影响及机理研究[D].沈阳:辽宁中医药大学,2016.
[2]Talman V,Ruskoaho H.Cardiac fibrosis in myocardial infarction—from repair and remodeling to regeneration[J].Cell Tissue Res,2016,365(3):563.
[3]Morrone D,Marzilli M.Role of RAAS inhibition in preventing left ventricular remodeling in patients post myocardial infarction[J].Heart Metab,2010(47):9.
[4]Roth L,Schrijvers D M,Martinet W,et al.AngiotensinⅡincreases coronary fibrosis,cardiac hypertrophy and the incidence of myocardial infarctions in Apo E-/-Fbn1C1039G+/-mice[J].Acta Cardiol,2016,71(4):483.
[5]Namsolleck P,Recarti C,Foulquier S,et al.AT(2)receptor and tissue injury:therapeutic implications.[J].Curr Hypertens Rep,2014,16(2):416.
[6]Balsinde J,Winstead M V,Dennis E A.Phospholipase A2,regulation of arachidonic acid mobilization[J].Febs Lett,2002,531(1):2.
[7]Wang J,Lu L,Wang Y,et al.Qishenyiqi dropping pill attenuates myocardial fibrosis in rats by inhibiting RAAS-mediated arachidonic acid inflammation[J].J Ethnopharmacol,2015,176(16):375.
[8]Li C,Wang J,Wang Q,et al.Qishen granules inhibit myocardial inflammation injury through regulating arachidonic acid metabolism[J].Sci Rep,2016,6:36949.
[9]Hsu C C,Lai S C.Matrix metalloproteinase-2,-9 and-13 are involved in fibronectin degradation of rat lung granulomatous fibrosis caused by Angiostrongylus cantonensis[J].Int J Exp Pathol,2007,88(6):437.
[10]Ito H,Duxbury M,Benoit E,et al.Fibronectin-induced COX-2mediates MMP-2 expression and invasiveness of rhabdomyosarcoma[J].Biochem Biophys Res Commun,2004,318(2):594.
[11]叶华.用代谢组学方法评价复方丹参片对血瘀动物的作用[D].西安:陕西师范大学,2011.
[12]杨暋慧,张暋浩,朱尔佳,等.复方丹参膜控缓释片与复方丹参片在Beagle犬体内的药动学及生物利用度的比较研究[J].今日药学,2014,24(6):393.
[13]潘桂湘.复方丹参方主要化学成分的研究[D].北京:北京中医药大学,2002.
[14]成娟,蔡文辉,朱云杰.肝苏颗粒联合复方丹参片治疗肝纤维化28例[J].中西医结合肝病杂志,2001,11(5):301.
[15]郭玉东,李丽,左泽平,等.复方丹参片对大鼠急性心肌缺血的保护作用[J].解放军药学学报,2016(6):497.
[16]Wang Y,Li C,Ouyang Y,et al.Cardioprotective effects of Qishenyiqi mediated by angiotensinⅡtype 1 receptor blockade and enhancing angiotensin-converting enzyme 2[J].Evid-based Complement Altern Med,2012,2012(6):978127.
[17]王蕾,赵明镜,杨涛,等.从心电图和超声心动图相关性分析研究心肌梗死后心衰模型的早期评价和筛选方法[J].中西医结合心脑血管病杂志,2017(22):2816.
[18]Amani M,Jeddi S,Ahmadiasl N,et al.Effect of HEMADO on level of CK-MB and LDH enzymes after ischemia/reperfusion injury in isolated rat heart[J].Bio Impacts,2013,3(2):101.
[19]De Mello W C.Intracellular angiotensinⅡdisrupts chemical communication and impairs metabolic cooperation between cardiac myocytes[J].Peptides,2015,72:57.
[20]Higuchi S,Ohtsu H,Suzuki H,et al.AngiotensinⅡsignal transduction through the AT1 receptor:novel insights into mechanisms and pathophysiology[J].Clin Sci,2007,112(8):417.
[21]Jenkins C M,Cedars A,Gross R W.Eicosanoid signalling pathways in the heart[J].Cardiovasc Res,2009,82(2):240.
[22]López B,González A,Querejeta R,et al.Myocardial COX-2overexpression and fibrosis in hypertensive heart disease.Role of angiotensinⅡ[J].Amer J Hypertens,2005,18(5):A233.
[23]Callejas N A,Casado M,Díazguerra M J,et al.Expression of cyclooxygenase-2 promotes the release of matrix metalloproteinase-2 and-9 in fetal rat hepatocytes[J].Hepatology,2001,33(4):860.
[24]Rhinehart K,Handelsman C A,Silldorff E P,et al.ANGⅡAT2 receptor modulates AT1 receptor-mediated descending vasa recta endothelial Ca2+signaling[J].Amer J Physiol Heart Circ Phy,2003,284(3):H779.
[25]马艳秋.丹参酮ⅡA对大鼠心脏纤维化作用及机制的研究[D].郑州:郑州大学,2016.
[26]张翼宙,卢冬冬,董颖,等.丹参酮ⅡA对大鼠肝纤维化的干预作用及其调控AngⅡ的分子机制[J].浙江中医药大学学报,2017,41(1):1.
[27]郑娴.人参皂苷Rb1对心衰大鼠心肌重构的影响及机理研究[D].沈阳:辽宁中医药大学,2016.