益母草碱的合成及其对心血管保护作用的研究
摘要
近年来,心血管疾病的发病率和致残致死率在全世界范围内呈明显上升趋势,其中缺血性心脏病是临床最常见的心血管疾病之一,缺血性心脏病是由于心脏冠状循环改变,引起冠状血流和心肌需求之间不平衡而导致的心肌损害。急性冠状动脉狭窄和闭塞可引起的急性心肌缺血甚至心肌梗死。心肌缺血(myocardialischemia,MI)治疗方法很多,其中药物治疗不失为一重要手段。虽然硝酸酯类、钙通道阻滞剂和β-受体阻断药等传统抗心肌缺血药物具有较好治疗作用,但均存在一定的不足之处,所以对抗心肌缺血药物的探索和抗心肌缺血机制的研究仍是一项重要的研究课题。由于中药的多靶点、多途径、副作用小等作用特点,因此对传统中药的研究越来越受到人们的重视,益母草为唇科益母草属植物,具有活血、祛瘀、调经、利水、清热解毒的功效,用于治疗月经不调、产后腹痛等。近年来,随着对其研究的逐渐增多,其药用价值、尤其在治疗心血管疾病方面的作用得到较深入的认识。我们的研究发现并证明了益母草的有效活性成分可被用于心功能的保护,并阐述了益母草有效活性成分的抗氧化机制。有文献报道,有效活性成分中的特异生物碱益母草碱能通过抑制外钙内流和内钙释放而发挥舒张血管平滑肌的作用,本研究进一步开展益母草中的有效成分益母草碱对心肌缺血的保护作用。
益母草碱在全草中的含量较少,分离提取得到的纯度也不高,因此,本实验探索出全新的益母草碱合成工艺,以丁香酸为起始原料,丁香酸的酚羟基用苄基保护并与中间体氨基丁醇进行缩合反应,建立了一条工艺简单、廉价、环保的全新合成益母草碱的途径,并经高效液相色谱分析测定纯度可达99.8%。
本实验用化学全合成的方法得到益母草碱,采用乳鼠心肌细胞缺氧、H9c2细胞系氧化应激、大鼠急性心肌梗死和心衰的体内和体外模型,用DNA电泳检测断裂的DNA片段,用RT-PCR和Western blot检测凋亡蛋白和信号分子来阐明益母草碱的心肌保护作用。
结果表明,益母草碱提高缺氧,双氧水(hydrogen peroxide,H_2O_2)损伤的心肌细胞的存活率及降低乳酸脱氢酶(lactate dehydrogenase,LDH)的漏出率,增加抗氧化酶的活力及直接消除活性氧簇(reactive oxygen species,ROS)作用,抑制心肌细胞凋亡,减轻细胞内钙超载,同时降低H_2O_2诱导的因ROS激活的线粒体凋亡途径,包括膜电位升高,细胞色素c的释放和Bax蛋白从胞浆到线粒体的转位,并且抑制caspase-3的激活及多聚酶(ADP-核糖)聚合酶(PARP)的断裂,发挥了对心肌细胞的保护作用。体内研究表明益母草碱能降低急性心肌梗死大鼠的梗死面积、降低血浆中的肌酸激酶(creatine kinase,CK)、LDH的含量、提高抗氧化酶活力并抑制心肌细胞凋亡,同时益母草碱能增大左室最大变化速率,改善收缩性能,舒张冠脉,使左室舒张末压降低,提高心衰大鼠的心功能,减轻肺淤血。
综上所述,我们的研究发现益母草碱对心肌细胞有直接的保护作用主要通过抗氧化,减轻细胞内钙超载,促进Bcl-2基因的表达和抑制Bax蛋白从胞浆到线粒体的转位发挥抑制线粒体凋亡途径,发挥了对缺血缺氧、ROS诱导的心肌细胞的损伤的保护作用。
In recent years, the incidence of cardiovascular disease mortality and disability all over the world has shown the obviously upward trend. Ischemic heart disease is one of the most common clinical cardiovascular diseases. Ischemic heart disease is myocardial damage for the reason of the imbalance between coronary blood flow and myocardial demand, causing by change of coronary circulation. Acute coronary artery stenosis and occlusion may lead to acute myocardial ischemia or myocardial infarction. There are a lot of treatments of myocardial ischemia, in which drug therapy may be one important means. Although some traditional anti-myocardial ischemia drugs, such as nitrates, calcium channel blockers andβ-receptor blockers, has good therapeutic effects, but there still are some deficiencies. Therefore, the exploration of anti-myocardial ischemia drugs and their mechanism remains an important research topic.
Traditional Chinese medicine causes more and more attention because of its small toxic side effects. Leonurus, which belongs to lips Section Motherwort Leonurus genus, has lots of effects, such as promoting blood flow, removing blood stasis, diuresis and clearing away heat and toxic. It's used for the treatment of irregular menstruation, postpartum abdominal pain, etc. In recent years, with the increase in its research, its medicinal value, particularly in the treatment of cardiovascular diseases has been more in-depth understood. Our studies have found that an effective active ingredient from Leonurus, Leonurine, can be used for the protection of cardiac function, and clarified its antioxidant mechanism. It has been reported that Leonurine can inhibit ectocellular calcium influx and intracellular calcium release and play a role in relaxation of vascular smooth muscle. Our study was to further discover the protective effect of Leonurine in myocardial ischemia.
As the minor amount of leonurine in Herb leonuri and the poor purity of its extracts, we explored synthesis of leonurine starting from syringic acid: the protecting of syringic acid's hydroxyl group by benzyl was followed by the condensation reaction between the resulting intermediate and amino-butanol. This is an economic and environment friendly route with 99.8% purity of final product identified by HPLC.
We took method of chemical total synthesis to get leonurine, and use model of hypoxic neonatal rat cardiomyocytes, oxidative stressed H9c2 cells, and rats with acute myocardial infracted and heart failure in vivo and vitro. We tested DNA breakage fragment using DNA gel electrophoresis, and the apoptosis proteins and signaling molecules using RT-PCR and Western blot, for further investigating the role of myocardial protection of Leonurine.
The results showed that Leonurine can enhance the viability of myocytes injuried by hypoxia or H_2O_2, reduce the leakage of LDH, increase the vitality of antioxidant enzymes and eliminate the effect of ROS, inhibit myocyte apoptosis, to reduce intracellular calcium overload. At the same time, it limited mitochondrial apoptosis due to ROS activation induced by H_2O_2, including increase of membrane potential, cytochrome c release and Bax protein from cytoplasm to mitochondrial translocation, and also inhibiting the activation of caspase-3 and broken of PARP. Leonurine in vivo study showed that it can reduce heart infarct size of acute myocardial ischemic rats, lower plasma CK, LDH levels, enhance the vitality of antioxidant enzymes and inhibit myocyte apoptosis, while Leonurine can increase the maximum changing rate of left ventricular, improve systolic performance, diastolic coronary artery, so that lower left ventricular end diastolic pressure, improve cardiac function and reduce pulmonary congestion in rats with heart failure.
As above, we found Leonurine has direct cardio-protective effect via antioxidant, lowering intracellular calcium overload, up-regulating Bcl-2 gene expression and inhibiting mitochondrial apoptosis by blocking Bax protein translocation from cytoplasm to mitochondrial, through which, it showed its protection to myocyte injury causing by hypoxia and ischemic induced by ROS.
益母草碱在全草中的含量较少,分离提取得到的纯度也不高,因此,本实验探索出全新的益母草碱合成工艺,以丁香酸为起始原料,丁香酸的酚羟基用苄基保护并与中间体氨基丁醇进行缩合反应,建立了一条工艺简单、廉价、环保的全新合成益母草碱的途径,并经高效液相色谱分析测定纯度可达99.8%。
本实验用化学全合成的方法得到益母草碱,采用乳鼠心肌细胞缺氧、H9c2细胞系氧化应激、大鼠急性心肌梗死和心衰的体内和体外模型,用DNA电泳检测断裂的DNA片段,用RT-PCR和Western blot检测凋亡蛋白和信号分子来阐明益母草碱的心肌保护作用。
结果表明,益母草碱提高缺氧,双氧水(hydrogen peroxide,H_2O_2)损伤的心肌细胞的存活率及降低乳酸脱氢酶(lactate dehydrogenase,LDH)的漏出率,增加抗氧化酶的活力及直接消除活性氧簇(reactive oxygen species,ROS)作用,抑制心肌细胞凋亡,减轻细胞内钙超载,同时降低H_2O_2诱导的因ROS激活的线粒体凋亡途径,包括膜电位升高,细胞色素c的释放和Bax蛋白从胞浆到线粒体的转位,并且抑制caspase-3的激活及多聚酶(ADP-核糖)聚合酶(PARP)的断裂,发挥了对心肌细胞的保护作用。体内研究表明益母草碱能降低急性心肌梗死大鼠的梗死面积、降低血浆中的肌酸激酶(creatine kinase,CK)、LDH的含量、提高抗氧化酶活力并抑制心肌细胞凋亡,同时益母草碱能增大左室最大变化速率,改善收缩性能,舒张冠脉,使左室舒张末压降低,提高心衰大鼠的心功能,减轻肺淤血。
综上所述,我们的研究发现益母草碱对心肌细胞有直接的保护作用主要通过抗氧化,减轻细胞内钙超载,促进Bcl-2基因的表达和抑制Bax蛋白从胞浆到线粒体的转位发挥抑制线粒体凋亡途径,发挥了对缺血缺氧、ROS诱导的心肌细胞的损伤的保护作用。
In recent years, the incidence of cardiovascular disease mortality and disability all over the world has shown the obviously upward trend. Ischemic heart disease is one of the most common clinical cardiovascular diseases. Ischemic heart disease is myocardial damage for the reason of the imbalance between coronary blood flow and myocardial demand, causing by change of coronary circulation. Acute coronary artery stenosis and occlusion may lead to acute myocardial ischemia or myocardial infarction. There are a lot of treatments of myocardial ischemia, in which drug therapy may be one important means. Although some traditional anti-myocardial ischemia drugs, such as nitrates, calcium channel blockers andβ-receptor blockers, has good therapeutic effects, but there still are some deficiencies. Therefore, the exploration of anti-myocardial ischemia drugs and their mechanism remains an important research topic.
Traditional Chinese medicine causes more and more attention because of its small toxic side effects. Leonurus, which belongs to lips Section Motherwort Leonurus genus, has lots of effects, such as promoting blood flow, removing blood stasis, diuresis and clearing away heat and toxic. It's used for the treatment of irregular menstruation, postpartum abdominal pain, etc. In recent years, with the increase in its research, its medicinal value, particularly in the treatment of cardiovascular diseases has been more in-depth understood. Our studies have found that an effective active ingredient from Leonurus, Leonurine, can be used for the protection of cardiac function, and clarified its antioxidant mechanism. It has been reported that Leonurine can inhibit ectocellular calcium influx and intracellular calcium release and play a role in relaxation of vascular smooth muscle. Our study was to further discover the protective effect of Leonurine in myocardial ischemia.
As the minor amount of leonurine in Herb leonuri and the poor purity of its extracts, we explored synthesis of leonurine starting from syringic acid: the protecting of syringic acid's hydroxyl group by benzyl was followed by the condensation reaction between the resulting intermediate and amino-butanol. This is an economic and environment friendly route with 99.8% purity of final product identified by HPLC.
We took method of chemical total synthesis to get leonurine, and use model of hypoxic neonatal rat cardiomyocytes, oxidative stressed H9c2 cells, and rats with acute myocardial infracted and heart failure in vivo and vitro. We tested DNA breakage fragment using DNA gel electrophoresis, and the apoptosis proteins and signaling molecules using RT-PCR and Western blot, for further investigating the role of myocardial protection of Leonurine.
The results showed that Leonurine can enhance the viability of myocytes injuried by hypoxia or H_2O_2, reduce the leakage of LDH, increase the vitality of antioxidant enzymes and eliminate the effect of ROS, inhibit myocyte apoptosis, to reduce intracellular calcium overload. At the same time, it limited mitochondrial apoptosis due to ROS activation induced by H_2O_2, including increase of membrane potential, cytochrome c release and Bax protein from cytoplasm to mitochondrial translocation, and also inhibiting the activation of caspase-3 and broken of PARP. Leonurine in vivo study showed that it can reduce heart infarct size of acute myocardial ischemic rats, lower plasma CK, LDH levels, enhance the vitality of antioxidant enzymes and inhibit myocyte apoptosis, while Leonurine can increase the maximum changing rate of left ventricular, improve systolic performance, diastolic coronary artery, so that lower left ventricular end diastolic pressure, improve cardiac function and reduce pulmonary congestion in rats with heart failure.
As above, we found Leonurine has direct cardio-protective effect via antioxidant, lowering intracellular calcium overload, up-regulating Bcl-2 gene expression and inhibiting mitochondrial apoptosis by blocking Bax protein translocation from cytoplasm to mitochondrial, through which, it showed its protection to myocyte injury causing by hypoxia and ischemic induced by ROS.
引文
[1]Zhu YZ,Zhu YC,Gohlke P,Unger T.Overview on pharmacological properties of angiotensin converting enzyme inhibitors[A].In:Hall & Unger eds.ACE Inhibition and Target Organ Protection[C].Oxford:Euromed Communications Ltd.1998:1-20.
[2]Hu S.A contribution to our knowledge of Leonurus L,i-mu-ts'ao,the Chinese motherwort[J].Am.J.Chin.Med.1976,4(3):219-237.
[3]Wang ZS,Li DW,Xia WJ,Qiu HQ,Zhu LY.The therapeutic effect of Herba leonuri in the treatment of coronary myocardial ischemia[J].J.Tradit.Chin.Med.1988,8(2):103-106.
[4]Chang CF,Li CZ.Experimental studies on the mechanism of anti-platelet aggregation action of Motherwort[J].Zhong Xi Yi Jie He Za Zhi.1986,6(1):39-40.
[5]Kuang PG,Zhou XF,Zhang FY,Lang SY.Motherwort and cerebral ischemia[J].J.Tradit.Chin.Med.1988,8(1):37-40.
[6]Zhu YZ,Huang SH,Tan BKH et al.Antioxidants in Chinese herbal medicine:a biochemical perspective[J].Nat.Prod.Rep.2004,21(4):478-489.
[7]Hu X,Mao JC,Huang SH,Sun J,Wu Y J,Wei D,On T,Zhu YZ.Protective effects of Herba Leonuri in ischemic models[A].In:Natural Products:A Must for Human Survive[C].Zhu YZ,Tan B,Bay BH,Liu CH.eds.London:World Scientific Press,2006:361-379.
[8]Sun J,Huang SH,Zhu YC et al.Anti-oxidative stress effects of Herba leonuri on ischemic rat hearts[J].Life.Sci.2005,76(26):3043-3056.
[9]Liu XH,Xin H,Zhu YZ.More than a mother-benefitingq herb:cardioprotective effect ofHerba leonuri[J].Acta Physiologica Sinica,2007,59(5):578-584.
[10]Chen CX,Kwan CY.Endothelium-independent vasorelaxation by leonurine,a plant alkaloid purified from Chinese motherwort[J].Life.Sci.2001,68(8):953-960.
[11]徐勇,智光,郑思新.益母草的心血管药理作用及临床应用[J].世界危重病杂志,2006,3(3):1312-1314.
[12]李素云,庞惠芳,卫洪昌,姜水印,朱晓梅.益母草生物碱和黄酮成分抗大鼠心肌缺血药效学研究[J].山东中医杂志,2006,2(25):114-116.
[13]Pang Shanling,Tsuchiyal Shizuko,Horiel Syunji,Uchida Masayuki,Murayamal Toshihiko,Kazuo Watanabe.Enhancement of phenylephrine-induced contraction in the isolated rat aorta with endothelium by H_2O-extract from an oriental medicinal plant Leonuri herba[J].Jpn.J.Pharmacol,2001,86(2):215-222.
[14]丛悦,王金辉,郭洪仁,等.益母草化学成分的分离与鉴定Ⅱ[J].中国药 物化学杂志,2003,13(6):349-352.
[15]张娴,鹏国平.益母草属化学成分研究进展[J].天然产物研究与开发,2003,15(2):162-166.
[16]丁伯平,熊莺,徐朝阳.益母草碱对急性血瘀症大鼠血流变学的影响[J].中国中医药科技,2004,11(1):36-37.
[17]周远鹏,刘文化,邵国贤.益母草碱、丁香酰胍醇及丁香酸氨基醇酯类化合物抗血小板聚集活性及其与结构的关系[J].中国药学杂志,1996,31(5):271-274.
[18]李万,蔡亚玲.益母草总生物碱的药理实验研究[J].华中科技大学学报,2002,31(2):168-170.
[19]Zhu RF,Ni SD,Wang XS.Study on new synthetic process for leonurine[J].Anhui university.Natural.Science.Edition.2005,29(4):84-86.
[20]阮金兰,杜俊蓉,曾庆忠,钱忠明.益母草的化学、药理和临床研究进展[J].中草药,2003,34(11):15-19.
[21]Bittira B,Kuang JQ,AI-Khaldi A,et al.In vitro preprogramming of marrow stromal cells for myocardial regeneration[J].Ann Thoral Surg.2002,74(4):1154-1160.
[22]Jeroudi MO,Hartley C J,Bolli R.Myocardial reperfusion injury:role of oxygen radicals and potential therapy with antioxidants[J].Am.J.Cardiol.1994,73(6):2-7.
[23]Zweier JL.Measurement of superoxide-derived free radicals in the reperfused heart:evidence for a free radical mechanism of reperfusion injury[J].J Biol Chem.1988,263(3):1353-1357.
[24]Peter MKang,Armin Haunstetter,Hiroki Aoki,Anny Usheva,Seigo Izumo.Morphological and molecular characterization of adult cardiomyocyte apoptosis during hypoxia and reoxygenation[J].Circ Res.2000,87(2):118-125.
[25]Rakhit RD,Edwards R J,Mockridge JW et al.Nitric oxide-induced cardioprotection in cultured rat ventricular myocytes[J].Am.J.Physiol.Heart.Circ.Physiol.2000,278(4):1211-1217.
[26]Dumay A,Rincheval V,Trotot P,Mignotte B,Vayssiere JL.The superoxide dismutase inhibitor diethyldithiocarbamate has antagonistic effects on apoptosis by triggering both cytochrome c release and caspase inhibition[J].Free Radic Biol Med.2006,40(8):1377-1390.
[27]MacLellan WR,Schneider MD.Death by design programmed cell death in cardiovascular biology and disease[J].Circ Res,1997,81(2):137-144.
[28]Kinugasa Y,Ogino K,Furuse Y,Shiomi T,Tsutsui H,Yamamoto T,Igawa O,Hisatome I,Shigemasa C.Allopurinol improves cardiac dysfunction after ischemia-reperfusion via reduction of oxidative stress in isolated perfused rat hearts [J].Circ J.2003,67(9):781-787.
[29]郑鸿翱,陈少如,尹俊。益母草对兔心肌缺血再灌注损伤时氧自由基的影响[J]。汕头大学医学院学报,1997,10(2):10-12.
[30]Netticadan T,Temsah R,Osada M,et al.Status of Ca~(2+)/calmodulin protein kinase phosphorylation of cardiac SR proteins in ischemia-reperfusion[J].Am.J.Physiol.1999,277:384-391.
[31]Thomas Netticadan,Rana M.Temsah,Kenichi Kawabata,et al.Ca~(2+)-overload inhibits the cardiac SR Ca~(2+)-calmodulin protein kinase activity[J].Biochemical and Biophysical Research Communications,2002,293(2):727-732.
[32]Shige matsu S,Arita M.Anoxia depresses sodium-calcium exchange currents in guinea-pig ventricular myocytes[J].J Mol Cell Cardiol,1999:31(4):895-906.
[33]Wattanapitayakul SK,Bauer JA.Oxidative pathways in cardiovascular disease:roles,mechanisms,and therapeutic implications[J].Pharmacol Ther.2001,89(2):187-206.
[34]Hinata M,Matsuoka I,lwamotoT,et al.Mechanism of Na~+/Ca~(2+) Exchanger Activation by Hydrogen Peroxide in Guinea-Pig Ventricular Myocytes[J].J Pharmacol Sci.2007,103(3):283-292.
[35]Sharikabad MN,Ostbye KM,Brors O.Effect of hydrogen peroxide on reoxygenation-induced Ca~(2+) accumulation in rat cardiomyocytes[J].Free Radic Biol Med.2004,37(4):531-538.
[36]Gao Z,Huang K,Xu H.Protective effects of flavonoids in the roots of Scutellaria baicalensis Georgi against hydrogen peroxide-induced oxidative stress in HS-SY5Y cells[J].Pharmacol Res.2001,43(2):173-178.
[37]Wang ZY,Cui M,Sun Lj.Angiopoietin-1 protects H9c2 cells from H2O2-induced apoptosis through AKT signaling.Biochemical and Biophysical Research Communications,2007,359(4):685-690.
[38]Ju YK,Saint DA,Gage PW.Hypoxia increase persistent sodium current rat ventricular myocytes[J].J Physiol.1996,497(2):337-347.
[39]Morin D,et al.Mitochondria as target for anti ischemic drugs[J].Adv Drug Deliv Rev.2001,49(12):151-174.
[40]Mark P.Mattson and Guido Kroemer.Mitochondrial in cell death:novel targets for neuroprotection and cardioprotection[J].Trends molecular Medicine,2003,9(5):196-204.
[41]余卫平,钱之玉,绪广林,等.西红华酸对心肌细胞氧化应激性损伤的作用[J].中国药科大学学报,2003,34(5):452-455.
[42]Long X,Goldenthal MJ,Wu GM,Marin-Garcia J.Mitochondrial Ca~(2+) flux and respiratory enzyme activity decline are early events in cardiomyocyte response to H_2O_2[J].J Mol Cell Cardiol.2004,37(1):63-70.
[43]Akao M,O'Rourke B,TeshimaY,Seharaseyon J,Marban E.Mechanistically distinct steps in the mitochondrial death pathway triggered by oxidative stress in cardiac myocytes[J].Circ Res.2003,92(2):186-194.
[44]Chen HW,Chien CT,Yu SL,Lee YT,Chen WJ.Cyclosporine A regulate oxidative stress-induced apoptosis in cardiomyocytes:mechanisms via ROS generation,iNOS and Hsp70[J].Brit J Pharmacol.2002,137(6):771-781.
[45]Brittsan AG,Kranias EG.Phospholamban and cardiac contractile function[J].J.Mol.Cell.Cardiol.2000,32(12):2131-2139.
[46]Mehmet H.Apoptosis:Caspases find a new place to hide[J].Nature,2000,403(6765):29-30.
[47]Borutaite V,Brown GC.Mitochondria in apoptosis of ischemic heart[J].FEBS Lett.2003,541(1-3):1-5.
[48]Turner NA,Xia F,Azhar G,Zhang X,Liu L,Wei JY.Oxidative stress induces DNA fragmentation and caspase activation via the c-Jun NH2-terminal kinase pathway in H9c2 cardiac muscle cells[J].J Mol Cell Cardiol.1998,30(9):1789-1801.
[49]Antonsson B,Montessuit S,Lauper S,Eskes R,Martinou JC.Bax oligomerization is required for channel-forming activity in liposomes and to trigger cytochrome crelease from mitochondria[J].Biochem J.2000,345(2):271-278.
[50]常青,王晓良.细胞色素C、线粒体与凋亡[J].中国药理学通报,2003,19(3):241-244.
[51]Li P,Nijhawan D,Budihardjo I,et al.Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade[J].Cell,1997,91(4):479-489
[52]Chang Q,Wang XL.Cytochrome C,mitochondria and apoptosis[J].Chin Pharm acol Bull.2003,19(3):241-244.
[53]Zou H,Li Y,Liu X,Wang X.An APAF21,cytochrome C multime2 ric complex is a functional apoptosome that activates procaspase[J].J Biol Chem.1999,274(17):11549-11556.
[54]Hu Y,BenedictM A,Ding L,Nunez G.Role of cytochrome C and dATP/ATP hydrolysis in Apaf212 mediated Caspase29 activation and apoptosis[J].EMBO J,1999,18(13):3586-3595.
[55]Saleh A,Srinivasula S M,Acharya S,et al.Cytochrome C and dATP mediated oligomerization of Apaf21 is a prerequisite for p ro2caspase29 activation[J].J B iol Chem.1999,274(25):17941-17945.
[56]Enari M,Sakahira H,Yokoyama H,et al.A Caspase2activated DNase that degrades DNA during apop tosis,and its inhibitor ICAD[J].Nature,1998,391(6662):43-50.
[57]Gottlieb RA,Engler RL.Apoptosis in myocardial isehemia-reperfusion[J].Ann N Y Acad Sci.1999,30(874):412-426.
[58]吴兆苏.心血管系统疾病流行病学及防治[M].北京:人民生出版社,2002.
[59]余志斌.心肌细胞凋亡在心力衰竭形成中的作用[J].生理科学进展,2000,31(3):265-268.
[60]Narula J,Haider N,Virmani R,et al.Apoptosis in myocytes in end-stage heart failure[J].N Engl J Med.1996,335(16):1182-1189.
[61]Olivetti G,Abbi R,Ouaini F,et al.Apoptosis in the failing human heart[J].N Engl J Med.1997,336(16):1131-1141.
[62]Olivetti G,Quaini F,Sala R,et al.Acute myocardial infarction in humans is associated with activation of programmed myocyte cell death in the surviving portion of the heart[J].J Mol Cell Cardiol.1996,28(16):2005-2016.
[63]Pries AR,Habazettl H,Ambrosio G.A review of methods for assessment of coronary microvascular disease in both clinical and experimental settings[J].Cardiovasc Res.2008,80(2):165-174.
[64]Park Channy,So Hong-Seob,Shin Chang-Ho,Baek Seung-Hwa,Moon Byung-Soon,Shin Sun-Ho.Quercetin protects the hydrogen peroxide-induced apoptosis via inhibition of mitochondrial dysfuntion in H9c2 cardiomyoblast cells.Biochem Pharmacol[J].2003,66(7):1287-1295.
[65]Mackay K,Mochly-Rosen D.Arachidonic acid protects neonatal rat cardiac myocytes from ischaemic injury through epsilon protein kinase C[J].Cardiovasc Res.2001,50(1):65-74.
[66]Kang N,Alexander G,Park JK,et al.Differential expression of protein kinase C isoforms in streptozotocin-induced diabetic rats[J].Kidney Int.1999,56(5):1737-1750.
[67]Dart RC,Sanders AB.Oxygen free radicals'and myocardial reperfusion injury[J].Ann Emerg Med.1988,17(1):53-58.
[68]Flitter WD.Free radicals and myocardial reperfusion injury[J].Br Med Bull.1993,49(3):545-555.
[69]McLeod LL,Sevanian A.Lipid peroxidation and modification of lipid composition in an endothelial cell model of ischemia and reperfusion[J].Free Radic Biol Med.1997,23(4):680-694.
[70]Vaux DL.Toward an understanding of the molecular mechanisms of physiological cell death[J].Proc Natl Acad Sci USA.1993,90(3):786-789.
[71]Qin F,Shite J,Mao W,et al.Selegiline atenuates cardiac oxidative stress and apoptosis in heart failure:association with improvement of cardiac function[J].Eur J Pharmacol.2003,461(3):149-158.
[72]Hofstaetter B,Taimor GS,Inserte J,et al.Inhibition of apoptotic responses after ischemic stress in isolated hearts and cardiomyocytes[J].Basic Res Cardiol.2002,97(6):479-488.
[73]Liu P,Xu B,Cavalieri TA,HoCPK CE.Age-related difference in myocardial function and inflammation in a rat model of myocardial ischemia-reperfusion[J].Cardiovasc Res.2002,56(3):443-453.
[74]Hochhauser E,Kivity S,Often D,et al.Bax ablation protects against myocardial ischemia-reperfusion injury in transgenic mice[J].Am J Physiol Heart Circ Physiol.2003,284(6):H2351-2359.
[75]Chang TH,Liu XY,Zhang XH,et al.Effects of dl-praeruptorin on interleukin-6level and Fas,bax,bcl-2 protein expression in ischemia—reperfusion myocardium[J].Acta Pharmacol Sin.2002,(9):769-774.
[76]Brocheriou V,Hagege AA,Oubenaissa A,et al.Cardiac functional improvement by a human Bcl-2 transgene in a mouse model of ischemia/reperfusion injury[J].J Gene Med.2000,2(5):326-333.
[77]Van P,De J.Myocyte hypertrophy and apoptosis:a balancing act[J].Cardiovasc Res.2004,63(3):487-499.
[78]Cesselli D,Jakoniuk I,Barlucchi L,et al.Oxidative stress—mediated cardiac cell death is a maior determinant of ventricular dysfunction and failure in dog dilated cardiomyopathy[J].Circ Res.2001,89(3):279-286.
[79]Hirotani S,Otsu K,Nishida K,et al.Involvement of nuclear factor,kappa B and apoptosis signal regulating kinase 1 in G-protein-coupled receptor agonist-induced cardiomyocyte hypertrophy[J].Circulation,2002,105(4):509-515.
[80]Swynghedauw B.Molecular mechanisms of myocardial remodeling[J].Physiol Rev.1999,79(1):215-262.
[81]Hunter JJ,Chien KR.Signaling pathways for cardiac hypertrophy and failure[J].N Engl J Med.1999,341(17):1276-1283.
[82]Ogata Y,Takahashi M,Uenos S,et al.Antiapoptotic effect of endothelin-1 in rat cardiomyocytes in vitro[J].Hypertension,2003,41(5):1156-1163.
[83]Bohn H,Martorana PA,Schonafinger K.Cardiovascular effects of the new nitric oxidedonor,pirsidomine.Hemodynamic profile and tolerance studies in anesthetized and consicious dogs[J].Fir J Pharmacol.1992,220(1):71-78.
[84]陈奇.中药药理研究方法学[M].北京:人民卫生出版社.1992:525-536.
[85]Grossamn W,Barry WH.Diastolic Pressure-volume relations in the diseased heart[J].Fed Proc.1980,39(2):148-155
[1] Zhu YZ, Zhu YC, Gohlke P, Unger T. ACE Inhibition and Target Organ Protection. In Hall & Unger, editors. Oxford, Euromed Communications Ltd., 1998, 1-20.
[2] Zhang X, Peng GP. Development of chemical constituents for Leonurus. Nat Prod Res Dev, 2003, 15(2): 162-164 (Chinese, English abstract).
[3] Zhu YZ, Zhu YC. Rediscovering remedies. Science Online highlighted in the regular issue of Science. 2002, 297(5585): 1231. http://intl-nextwave.sciencemag.Org/cgi/content/full/2002/08/21 /10
[4] Dan B, Andrew G. Herbs that invigorate the blood. In: Chinese Herbal Medicine, Materia Medica. Seattle: Eastland Press, 1993, 273-274.
[5] Zhu YP. Chinese Materia Medica: Chemistry, Pharmacology and Applications. London: Harwood Academic Publishers, 1998, 466-468.
[6] Chang IM. Anti-aging and health-promoting constituents derived from traditional oriental herbal remedies: information retrieval using the TradiMed 2000 DB. Ann NY Acad Sci, 2001, 928: 281-286.
[7] Ruan JL, Du JR, Zeng QZ, Qian ZM. Advance in studies on chemical components,pharmacological effect and clinical application of Leonurus japonicus. Chin Trad Herb Drugs, 2003, 34(11): 15-19.
[8] Yin J, Wang HL. Effect on blood rheology and thrombosis in rats myocardial ischemia by motherwort herb. Chin J Thromb Hemos, 2001, 7(1): 13-15.
[9] Zhang CF, Zhu XM, Gong B. Study on mechanism of anti-platelet aggregation by motherwort herb. Chin J Integ Tradit West Med, 1986, 1: 39-40.
[10] Zou QJ, Bi RA, Li JM, Feng JB, Yu AM, Chan HP, Zhen MX. Effect of motherwort on blood hyperviscosity. Shanxi Med J, 1989, 1: 13-14.
[11] Sun J, Huang SH, Zhu YC, Whiteman M, Wang MJ, Tan BK, Zhu YZ. Anti-oxidative stress effects of Herba leonuri on ischemic rat hearts. Life Sci, 2005,76: 3043-3056.
[12] Wang XG, Zhu ZY. Studies on the essential constituents of three plants from Leonurus spp. J Chin Med Mater, 1991, 14(11): 35-36.
[13] Bing RJ. Some aspects of biochemistry of myocardial infarction. Cell Mol Life Sci, 2001, 58: 351-355
[14] Huang JY, Dong RS, Yan JZ. Therapeutic mechanisms of Leonurus Hererpophyllus treatment for coronary heart disease from changes of microcirculation hemorheology. Chin J Hemorh, 1995, 5(3): 16-19.
[15] Dong RS, Chen SR, Huang JY, Yan JZ, Hong SH, Cai JS. Clinical observation of silent myocardial ischemia treatment with Leonurus Hererpophyllus injection. J Shantou Univ Med Coll, 1998,11(1): 6-7.
[16] Li SY, Jiang SY, Wei HC. Pharmacodynamic research on alkaloid and chromocor of Leonurus Heteropyllus Sweet in isoproterenol-induced myocardial injury in rats. Acta Univ Tradit Med Sin Pharmacol Shanghai, 2006, 20 (1): 61-63.
[17] Wang YS. The Application and Pharmacology of Chinese Medicine. 1st ed. Beijing: People's Health Publisher, 1983, 916-922.
[18] Wang ZS, Li DW, Xia WJ, Qiu HQ, Zhu LY. The therapeutic effect of herba Leonuri in the treatment of coronary myocardial ischemia. J Tradit Chin Med, 1988, 8(2): 103-106.
[19] Zhang CF, Jia YS, Wei HC, Zhu XM, Hui YM, Zhang CY, Mo QZ, Gong B. Studies on actions of extract of motherwort. J Tradit Chin Med, 1982, 2(4): 267-270.
[20] Jiang SY, Huang PX, Wei HC, Zhang C, Wang JH, Wu LQ. Effects of alkaloid of Leonurus Heteropyllus Sweet on heart function after myocardial infarction in rats. Shanghai J Tradit Chin Med, 2006,40(10): 53-56.
[21] Chen SR, Chen Sui, Zheng HG, Chen HQ. Comparison the therapeutical effects of Leonuru Heterophyllus injection and composite Salvia Miltiorrhiza injection on myocardial ischemiareperfusion injury in rabbits and on the patients with coronary heart disease. J Shantou Univ Med Coll, 2003; 16 (1): 9-11.
[22] Zheng HA, Chen Sui, Chen SR. Comparison protection effect of Leonuru Heterophyllus and Chinese Angelica injection on rabbit myocardial ischemia-reperfusion injury. Chin J Lab Anim Sci, 2002,12 (6): 384-384.
[23] Chen SR, Chen Sui, Li QY. The therapeutic effects of herba leonuri injection on arrhythmia in patients with coronary heart diseases and in rats with myocardial ischemia reperfusion injury. J Clin Cardiol, 2002,18(10): 490-491.
[24] Chen CX, Kwan CY. Endothelium-independent vasorelaxation by leonurine, a plant alkaloid purified from Chinese motherwort. Life Sci 2001, 68: 953-960.
[25] Wattanapitayakul SK, Bauer JA. Oxidative pathways in cardiovascular disease: roles, mechanisms, and therapeutic implications. Pharmacol Ther 2001, 89: 187-206.
[26] Ji XY, Tan BKH, Huang SH, Whiteman M, Zhu YC, Wu JY, Wei D, Zhu YZ. Effects of Salvia miltiorrhiza in ischemic myocardium in experimental rats. In:Perspective: Novel Compounds from Natural Products in New Millenniums. Tan B, Bay BH, Zhu YZ. eds. London: World Scientific Press, 2004,152-162.
[27] Zhu YZ, Huang SH, Tan BKH, Sun J, Whiteman M, Zhu YC. Antioxidants in Chinese herbal medicines: a biochemical perspective. Nat Prod Rep 2004, 21:478-489.
[28] Dhalla NS, Temsah RM, Netticadan T. Role of oxidative stress in cardiovascular diseases. J Hypertens, 2000, 18: 655-673.
[29] Yasmin W, Strynadka KD, Schulz R. Generation of peroxynitrite contributes to ischemia-reperfusion injury in isolated rat hearts. Cardiovasc Res, 1997, 33: 422-432.
[30] Rimm EB, Stampfer MJ. Antioxidants for vascular disease. Med Clin North Am, 2000, 84: 239-249.
[31] Zheng HA, Chen SR, Yin Jun. Effect of Leonurus Heterophyllus on oxygen free radicals during myocardial ischemia reperfusion injury in rabbits. J Shantou Univ Med Coll, 1997, 10(2): 10-12.
[32] Best PJ, Hasdai D, Sangiorgi G, Schwartz RS, Holmes DR Jr, Simari RD, Lerman A. Apoptosis-basic concepts and implications in coronary artery disease.Arterioscler Thromb Vase Biol, 1999,19: 14-22.
[33] Ferrari R, Agnoletti L, Comini L, Gaia G, Bachetti T, Cargnoni A, Ceconi C,Curello S, Visioli O. Oxidative stress during myocardial ischemia and heart failure. Eur Heart J, 1998,19:B2-B11.
[34] Turner NA, Xia F, Azhar G, Zhang XM, Liu LX. Oxidative stress induces DNA fragmentation and caspase activation via the JNK pathway in H9c2 cardiac muscle cells. J Mol Cell Cardiol, 1998, 30: 1789-1801.
[35] Hu X, Mao JC, Huang SH, Sun J, Wu YJ, Wei D, On T, Zhu YZ. Protective effects of Herba Leonuri in ischemic models. In: Natural Products: A Must for Human Survive. Zhu YZ, Tan B, Bay BH, Liu CH. eds. London: World Scientific Press, 2006, 361-379.
[36] Wang ZS, Li DW, Xia WJ, Qiu HQ, Zhu LY. The therapeutic effect of Herba Leonuri in the treatment of coronary myocardial ischemia. J Tradit Chin Med, 1988,8(2): 103-106.
[37] Chen HJ, Li DY, Saldeen T, Romeo F, Mehta JL. Mixed tocopherol preparation is superior to a-tocopherol alone against hypoxia-reoxygenation injury. Biochem Biophy Res Com, 2002, 291: 349-353.
[38] Loh KP, Low LS, Wong WH, Chou WH, Zhu YZ. Molecular mechanisms of cerebral protection from Ginkgo biloba extract on stroked rats. 2006; 398(1-2): 28-33.
[39] Hudlicka O, Brown M, Egginton S. Angiogenesis in skeletal and cardiac muscle. Physiol Rev, 1992, 72: 369-417.
[40] Schaper W. New paradigms for collateral vessel growth. Basic Res Cardiol, 1993, 88: 193-198.
[41] Waltenberger J. Modulation of growth factor action: implications for the treatment of cardiovascular diseases. Circulation, 1997, 96: 4083-4094.
[42] Li DW, Wang ZS, Qiu HQ. Herba Leonuri treatment for 80 patients with coronary heart diseases. Shanxi J Tradit Chin Med, 1984, 5 (10): 11-12.
[43] Zheng GW, Chen SR. Clinical observation of Herba Leonuri pill treatment for patients with myocardial ischemia. Chin J Inf Tradit Chin Med, 2000, 7(2): 19.
[44] Yan JZ, Chen SR. Curative effects of Leonuru Hererophyllus on coronary heart diseases. J Shantou Univ Med Coll, 2000, 13(1): 20-22.
[2]Hu S.A contribution to our knowledge of Leonurus L,i-mu-ts'ao,the Chinese motherwort[J].Am.J.Chin.Med.1976,4(3):219-237.
[3]Wang ZS,Li DW,Xia WJ,Qiu HQ,Zhu LY.The therapeutic effect of Herba leonuri in the treatment of coronary myocardial ischemia[J].J.Tradit.Chin.Med.1988,8(2):103-106.
[4]Chang CF,Li CZ.Experimental studies on the mechanism of anti-platelet aggregation action of Motherwort[J].Zhong Xi Yi Jie He Za Zhi.1986,6(1):39-40.
[5]Kuang PG,Zhou XF,Zhang FY,Lang SY.Motherwort and cerebral ischemia[J].J.Tradit.Chin.Med.1988,8(1):37-40.
[6]Zhu YZ,Huang SH,Tan BKH et al.Antioxidants in Chinese herbal medicine:a biochemical perspective[J].Nat.Prod.Rep.2004,21(4):478-489.
[7]Hu X,Mao JC,Huang SH,Sun J,Wu Y J,Wei D,On T,Zhu YZ.Protective effects of Herba Leonuri in ischemic models[A].In:Natural Products:A Must for Human Survive[C].Zhu YZ,Tan B,Bay BH,Liu CH.eds.London:World Scientific Press,2006:361-379.
[8]Sun J,Huang SH,Zhu YC et al.Anti-oxidative stress effects of Herba leonuri on ischemic rat hearts[J].Life.Sci.2005,76(26):3043-3056.
[9]Liu XH,Xin H,Zhu YZ.More than a mother-benefitingq herb:cardioprotective effect ofHerba leonuri[J].Acta Physiologica Sinica,2007,59(5):578-584.
[10]Chen CX,Kwan CY.Endothelium-independent vasorelaxation by leonurine,a plant alkaloid purified from Chinese motherwort[J].Life.Sci.2001,68(8):953-960.
[11]徐勇,智光,郑思新.益母草的心血管药理作用及临床应用[J].世界危重病杂志,2006,3(3):1312-1314.
[12]李素云,庞惠芳,卫洪昌,姜水印,朱晓梅.益母草生物碱和黄酮成分抗大鼠心肌缺血药效学研究[J].山东中医杂志,2006,2(25):114-116.
[13]Pang Shanling,Tsuchiyal Shizuko,Horiel Syunji,Uchida Masayuki,Murayamal Toshihiko,Kazuo Watanabe.Enhancement of phenylephrine-induced contraction in the isolated rat aorta with endothelium by H_2O-extract from an oriental medicinal plant Leonuri herba[J].Jpn.J.Pharmacol,2001,86(2):215-222.
[14]丛悦,王金辉,郭洪仁,等.益母草化学成分的分离与鉴定Ⅱ[J].中国药 物化学杂志,2003,13(6):349-352.
[15]张娴,鹏国平.益母草属化学成分研究进展[J].天然产物研究与开发,2003,15(2):162-166.
[16]丁伯平,熊莺,徐朝阳.益母草碱对急性血瘀症大鼠血流变学的影响[J].中国中医药科技,2004,11(1):36-37.
[17]周远鹏,刘文化,邵国贤.益母草碱、丁香酰胍醇及丁香酸氨基醇酯类化合物抗血小板聚集活性及其与结构的关系[J].中国药学杂志,1996,31(5):271-274.
[18]李万,蔡亚玲.益母草总生物碱的药理实验研究[J].华中科技大学学报,2002,31(2):168-170.
[19]Zhu RF,Ni SD,Wang XS.Study on new synthetic process for leonurine[J].Anhui university.Natural.Science.Edition.2005,29(4):84-86.
[20]阮金兰,杜俊蓉,曾庆忠,钱忠明.益母草的化学、药理和临床研究进展[J].中草药,2003,34(11):15-19.
[21]Bittira B,Kuang JQ,AI-Khaldi A,et al.In vitro preprogramming of marrow stromal cells for myocardial regeneration[J].Ann Thoral Surg.2002,74(4):1154-1160.
[22]Jeroudi MO,Hartley C J,Bolli R.Myocardial reperfusion injury:role of oxygen radicals and potential therapy with antioxidants[J].Am.J.Cardiol.1994,73(6):2-7.
[23]Zweier JL.Measurement of superoxide-derived free radicals in the reperfused heart:evidence for a free radical mechanism of reperfusion injury[J].J Biol Chem.1988,263(3):1353-1357.
[24]Peter MKang,Armin Haunstetter,Hiroki Aoki,Anny Usheva,Seigo Izumo.Morphological and molecular characterization of adult cardiomyocyte apoptosis during hypoxia and reoxygenation[J].Circ Res.2000,87(2):118-125.
[25]Rakhit RD,Edwards R J,Mockridge JW et al.Nitric oxide-induced cardioprotection in cultured rat ventricular myocytes[J].Am.J.Physiol.Heart.Circ.Physiol.2000,278(4):1211-1217.
[26]Dumay A,Rincheval V,Trotot P,Mignotte B,Vayssiere JL.The superoxide dismutase inhibitor diethyldithiocarbamate has antagonistic effects on apoptosis by triggering both cytochrome c release and caspase inhibition[J].Free Radic Biol Med.2006,40(8):1377-1390.
[27]MacLellan WR,Schneider MD.Death by design programmed cell death in cardiovascular biology and disease[J].Circ Res,1997,81(2):137-144.
[28]Kinugasa Y,Ogino K,Furuse Y,Shiomi T,Tsutsui H,Yamamoto T,Igawa O,Hisatome I,Shigemasa C.Allopurinol improves cardiac dysfunction after ischemia-reperfusion via reduction of oxidative stress in isolated perfused rat hearts [J].Circ J.2003,67(9):781-787.
[29]郑鸿翱,陈少如,尹俊。益母草对兔心肌缺血再灌注损伤时氧自由基的影响[J]。汕头大学医学院学报,1997,10(2):10-12.
[30]Netticadan T,Temsah R,Osada M,et al.Status of Ca~(2+)/calmodulin protein kinase phosphorylation of cardiac SR proteins in ischemia-reperfusion[J].Am.J.Physiol.1999,277:384-391.
[31]Thomas Netticadan,Rana M.Temsah,Kenichi Kawabata,et al.Ca~(2+)-overload inhibits the cardiac SR Ca~(2+)-calmodulin protein kinase activity[J].Biochemical and Biophysical Research Communications,2002,293(2):727-732.
[32]Shige matsu S,Arita M.Anoxia depresses sodium-calcium exchange currents in guinea-pig ventricular myocytes[J].J Mol Cell Cardiol,1999:31(4):895-906.
[33]Wattanapitayakul SK,Bauer JA.Oxidative pathways in cardiovascular disease:roles,mechanisms,and therapeutic implications[J].Pharmacol Ther.2001,89(2):187-206.
[34]Hinata M,Matsuoka I,lwamotoT,et al.Mechanism of Na~+/Ca~(2+) Exchanger Activation by Hydrogen Peroxide in Guinea-Pig Ventricular Myocytes[J].J Pharmacol Sci.2007,103(3):283-292.
[35]Sharikabad MN,Ostbye KM,Brors O.Effect of hydrogen peroxide on reoxygenation-induced Ca~(2+) accumulation in rat cardiomyocytes[J].Free Radic Biol Med.2004,37(4):531-538.
[36]Gao Z,Huang K,Xu H.Protective effects of flavonoids in the roots of Scutellaria baicalensis Georgi against hydrogen peroxide-induced oxidative stress in HS-SY5Y cells[J].Pharmacol Res.2001,43(2):173-178.
[37]Wang ZY,Cui M,Sun Lj.Angiopoietin-1 protects H9c2 cells from H2O2-induced apoptosis through AKT signaling.Biochemical and Biophysical Research Communications,2007,359(4):685-690.
[38]Ju YK,Saint DA,Gage PW.Hypoxia increase persistent sodium current rat ventricular myocytes[J].J Physiol.1996,497(2):337-347.
[39]Morin D,et al.Mitochondria as target for anti ischemic drugs[J].Adv Drug Deliv Rev.2001,49(12):151-174.
[40]Mark P.Mattson and Guido Kroemer.Mitochondrial in cell death:novel targets for neuroprotection and cardioprotection[J].Trends molecular Medicine,2003,9(5):196-204.
[41]余卫平,钱之玉,绪广林,等.西红华酸对心肌细胞氧化应激性损伤的作用[J].中国药科大学学报,2003,34(5):452-455.
[42]Long X,Goldenthal MJ,Wu GM,Marin-Garcia J.Mitochondrial Ca~(2+) flux and respiratory enzyme activity decline are early events in cardiomyocyte response to H_2O_2[J].J Mol Cell Cardiol.2004,37(1):63-70.
[43]Akao M,O'Rourke B,TeshimaY,Seharaseyon J,Marban E.Mechanistically distinct steps in the mitochondrial death pathway triggered by oxidative stress in cardiac myocytes[J].Circ Res.2003,92(2):186-194.
[44]Chen HW,Chien CT,Yu SL,Lee YT,Chen WJ.Cyclosporine A regulate oxidative stress-induced apoptosis in cardiomyocytes:mechanisms via ROS generation,iNOS and Hsp70[J].Brit J Pharmacol.2002,137(6):771-781.
[45]Brittsan AG,Kranias EG.Phospholamban and cardiac contractile function[J].J.Mol.Cell.Cardiol.2000,32(12):2131-2139.
[46]Mehmet H.Apoptosis:Caspases find a new place to hide[J].Nature,2000,403(6765):29-30.
[47]Borutaite V,Brown GC.Mitochondria in apoptosis of ischemic heart[J].FEBS Lett.2003,541(1-3):1-5.
[48]Turner NA,Xia F,Azhar G,Zhang X,Liu L,Wei JY.Oxidative stress induces DNA fragmentation and caspase activation via the c-Jun NH2-terminal kinase pathway in H9c2 cardiac muscle cells[J].J Mol Cell Cardiol.1998,30(9):1789-1801.
[49]Antonsson B,Montessuit S,Lauper S,Eskes R,Martinou JC.Bax oligomerization is required for channel-forming activity in liposomes and to trigger cytochrome crelease from mitochondria[J].Biochem J.2000,345(2):271-278.
[50]常青,王晓良.细胞色素C、线粒体与凋亡[J].中国药理学通报,2003,19(3):241-244.
[51]Li P,Nijhawan D,Budihardjo I,et al.Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade[J].Cell,1997,91(4):479-489
[52]Chang Q,Wang XL.Cytochrome C,mitochondria and apoptosis[J].Chin Pharm acol Bull.2003,19(3):241-244.
[53]Zou H,Li Y,Liu X,Wang X.An APAF21,cytochrome C multime2 ric complex is a functional apoptosome that activates procaspase[J].J Biol Chem.1999,274(17):11549-11556.
[54]Hu Y,BenedictM A,Ding L,Nunez G.Role of cytochrome C and dATP/ATP hydrolysis in Apaf212 mediated Caspase29 activation and apoptosis[J].EMBO J,1999,18(13):3586-3595.
[55]Saleh A,Srinivasula S M,Acharya S,et al.Cytochrome C and dATP mediated oligomerization of Apaf21 is a prerequisite for p ro2caspase29 activation[J].J B iol Chem.1999,274(25):17941-17945.
[56]Enari M,Sakahira H,Yokoyama H,et al.A Caspase2activated DNase that degrades DNA during apop tosis,and its inhibitor ICAD[J].Nature,1998,391(6662):43-50.
[57]Gottlieb RA,Engler RL.Apoptosis in myocardial isehemia-reperfusion[J].Ann N Y Acad Sci.1999,30(874):412-426.
[58]吴兆苏.心血管系统疾病流行病学及防治[M].北京:人民生出版社,2002.
[59]余志斌.心肌细胞凋亡在心力衰竭形成中的作用[J].生理科学进展,2000,31(3):265-268.
[60]Narula J,Haider N,Virmani R,et al.Apoptosis in myocytes in end-stage heart failure[J].N Engl J Med.1996,335(16):1182-1189.
[61]Olivetti G,Abbi R,Ouaini F,et al.Apoptosis in the failing human heart[J].N Engl J Med.1997,336(16):1131-1141.
[62]Olivetti G,Quaini F,Sala R,et al.Acute myocardial infarction in humans is associated with activation of programmed myocyte cell death in the surviving portion of the heart[J].J Mol Cell Cardiol.1996,28(16):2005-2016.
[63]Pries AR,Habazettl H,Ambrosio G.A review of methods for assessment of coronary microvascular disease in both clinical and experimental settings[J].Cardiovasc Res.2008,80(2):165-174.
[64]Park Channy,So Hong-Seob,Shin Chang-Ho,Baek Seung-Hwa,Moon Byung-Soon,Shin Sun-Ho.Quercetin protects the hydrogen peroxide-induced apoptosis via inhibition of mitochondrial dysfuntion in H9c2 cardiomyoblast cells.Biochem Pharmacol[J].2003,66(7):1287-1295.
[65]Mackay K,Mochly-Rosen D.Arachidonic acid protects neonatal rat cardiac myocytes from ischaemic injury through epsilon protein kinase C[J].Cardiovasc Res.2001,50(1):65-74.
[66]Kang N,Alexander G,Park JK,et al.Differential expression of protein kinase C isoforms in streptozotocin-induced diabetic rats[J].Kidney Int.1999,56(5):1737-1750.
[67]Dart RC,Sanders AB.Oxygen free radicals'and myocardial reperfusion injury[J].Ann Emerg Med.1988,17(1):53-58.
[68]Flitter WD.Free radicals and myocardial reperfusion injury[J].Br Med Bull.1993,49(3):545-555.
[69]McLeod LL,Sevanian A.Lipid peroxidation and modification of lipid composition in an endothelial cell model of ischemia and reperfusion[J].Free Radic Biol Med.1997,23(4):680-694.
[70]Vaux DL.Toward an understanding of the molecular mechanisms of physiological cell death[J].Proc Natl Acad Sci USA.1993,90(3):786-789.
[71]Qin F,Shite J,Mao W,et al.Selegiline atenuates cardiac oxidative stress and apoptosis in heart failure:association with improvement of cardiac function[J].Eur J Pharmacol.2003,461(3):149-158.
[72]Hofstaetter B,Taimor GS,Inserte J,et al.Inhibition of apoptotic responses after ischemic stress in isolated hearts and cardiomyocytes[J].Basic Res Cardiol.2002,97(6):479-488.
[73]Liu P,Xu B,Cavalieri TA,HoCPK CE.Age-related difference in myocardial function and inflammation in a rat model of myocardial ischemia-reperfusion[J].Cardiovasc Res.2002,56(3):443-453.
[74]Hochhauser E,Kivity S,Often D,et al.Bax ablation protects against myocardial ischemia-reperfusion injury in transgenic mice[J].Am J Physiol Heart Circ Physiol.2003,284(6):H2351-2359.
[75]Chang TH,Liu XY,Zhang XH,et al.Effects of dl-praeruptorin on interleukin-6level and Fas,bax,bcl-2 protein expression in ischemia—reperfusion myocardium[J].Acta Pharmacol Sin.2002,(9):769-774.
[76]Brocheriou V,Hagege AA,Oubenaissa A,et al.Cardiac functional improvement by a human Bcl-2 transgene in a mouse model of ischemia/reperfusion injury[J].J Gene Med.2000,2(5):326-333.
[77]Van P,De J.Myocyte hypertrophy and apoptosis:a balancing act[J].Cardiovasc Res.2004,63(3):487-499.
[78]Cesselli D,Jakoniuk I,Barlucchi L,et al.Oxidative stress—mediated cardiac cell death is a maior determinant of ventricular dysfunction and failure in dog dilated cardiomyopathy[J].Circ Res.2001,89(3):279-286.
[79]Hirotani S,Otsu K,Nishida K,et al.Involvement of nuclear factor,kappa B and apoptosis signal regulating kinase 1 in G-protein-coupled receptor agonist-induced cardiomyocyte hypertrophy[J].Circulation,2002,105(4):509-515.
[80]Swynghedauw B.Molecular mechanisms of myocardial remodeling[J].Physiol Rev.1999,79(1):215-262.
[81]Hunter JJ,Chien KR.Signaling pathways for cardiac hypertrophy and failure[J].N Engl J Med.1999,341(17):1276-1283.
[82]Ogata Y,Takahashi M,Uenos S,et al.Antiapoptotic effect of endothelin-1 in rat cardiomyocytes in vitro[J].Hypertension,2003,41(5):1156-1163.
[83]Bohn H,Martorana PA,Schonafinger K.Cardiovascular effects of the new nitric oxidedonor,pirsidomine.Hemodynamic profile and tolerance studies in anesthetized and consicious dogs[J].Fir J Pharmacol.1992,220(1):71-78.
[84]陈奇.中药药理研究方法学[M].北京:人民卫生出版社.1992:525-536.
[85]Grossamn W,Barry WH.Diastolic Pressure-volume relations in the diseased heart[J].Fed Proc.1980,39(2):148-155
[1] Zhu YZ, Zhu YC, Gohlke P, Unger T. ACE Inhibition and Target Organ Protection. In Hall & Unger, editors. Oxford, Euromed Communications Ltd., 1998, 1-20.
[2] Zhang X, Peng GP. Development of chemical constituents for Leonurus. Nat Prod Res Dev, 2003, 15(2): 162-164 (Chinese, English abstract).
[3] Zhu YZ, Zhu YC. Rediscovering remedies. Science Online highlighted in the regular issue of Science. 2002, 297(5585): 1231. http://intl-nextwave.sciencemag.Org/cgi/content/full/2002/08/21 /10
[4] Dan B, Andrew G. Herbs that invigorate the blood. In: Chinese Herbal Medicine, Materia Medica. Seattle: Eastland Press, 1993, 273-274.
[5] Zhu YP. Chinese Materia Medica: Chemistry, Pharmacology and Applications. London: Harwood Academic Publishers, 1998, 466-468.
[6] Chang IM. Anti-aging and health-promoting constituents derived from traditional oriental herbal remedies: information retrieval using the TradiMed 2000 DB. Ann NY Acad Sci, 2001, 928: 281-286.
[7] Ruan JL, Du JR, Zeng QZ, Qian ZM. Advance in studies on chemical components,pharmacological effect and clinical application of Leonurus japonicus. Chin Trad Herb Drugs, 2003, 34(11): 15-19.
[8] Yin J, Wang HL. Effect on blood rheology and thrombosis in rats myocardial ischemia by motherwort herb. Chin J Thromb Hemos, 2001, 7(1): 13-15.
[9] Zhang CF, Zhu XM, Gong B. Study on mechanism of anti-platelet aggregation by motherwort herb. Chin J Integ Tradit West Med, 1986, 1: 39-40.
[10] Zou QJ, Bi RA, Li JM, Feng JB, Yu AM, Chan HP, Zhen MX. Effect of motherwort on blood hyperviscosity. Shanxi Med J, 1989, 1: 13-14.
[11] Sun J, Huang SH, Zhu YC, Whiteman M, Wang MJ, Tan BK, Zhu YZ. Anti-oxidative stress effects of Herba leonuri on ischemic rat hearts. Life Sci, 2005,76: 3043-3056.
[12] Wang XG, Zhu ZY. Studies on the essential constituents of three plants from Leonurus spp. J Chin Med Mater, 1991, 14(11): 35-36.
[13] Bing RJ. Some aspects of biochemistry of myocardial infarction. Cell Mol Life Sci, 2001, 58: 351-355
[14] Huang JY, Dong RS, Yan JZ. Therapeutic mechanisms of Leonurus Hererpophyllus treatment for coronary heart disease from changes of microcirculation hemorheology. Chin J Hemorh, 1995, 5(3): 16-19.
[15] Dong RS, Chen SR, Huang JY, Yan JZ, Hong SH, Cai JS. Clinical observation of silent myocardial ischemia treatment with Leonurus Hererpophyllus injection. J Shantou Univ Med Coll, 1998,11(1): 6-7.
[16] Li SY, Jiang SY, Wei HC. Pharmacodynamic research on alkaloid and chromocor of Leonurus Heteropyllus Sweet in isoproterenol-induced myocardial injury in rats. Acta Univ Tradit Med Sin Pharmacol Shanghai, 2006, 20 (1): 61-63.
[17] Wang YS. The Application and Pharmacology of Chinese Medicine. 1st ed. Beijing: People's Health Publisher, 1983, 916-922.
[18] Wang ZS, Li DW, Xia WJ, Qiu HQ, Zhu LY. The therapeutic effect of herba Leonuri in the treatment of coronary myocardial ischemia. J Tradit Chin Med, 1988, 8(2): 103-106.
[19] Zhang CF, Jia YS, Wei HC, Zhu XM, Hui YM, Zhang CY, Mo QZ, Gong B. Studies on actions of extract of motherwort. J Tradit Chin Med, 1982, 2(4): 267-270.
[20] Jiang SY, Huang PX, Wei HC, Zhang C, Wang JH, Wu LQ. Effects of alkaloid of Leonurus Heteropyllus Sweet on heart function after myocardial infarction in rats. Shanghai J Tradit Chin Med, 2006,40(10): 53-56.
[21] Chen SR, Chen Sui, Zheng HG, Chen HQ. Comparison the therapeutical effects of Leonuru Heterophyllus injection and composite Salvia Miltiorrhiza injection on myocardial ischemiareperfusion injury in rabbits and on the patients with coronary heart disease. J Shantou Univ Med Coll, 2003; 16 (1): 9-11.
[22] Zheng HA, Chen Sui, Chen SR. Comparison protection effect of Leonuru Heterophyllus and Chinese Angelica injection on rabbit myocardial ischemia-reperfusion injury. Chin J Lab Anim Sci, 2002,12 (6): 384-384.
[23] Chen SR, Chen Sui, Li QY. The therapeutic effects of herba leonuri injection on arrhythmia in patients with coronary heart diseases and in rats with myocardial ischemia reperfusion injury. J Clin Cardiol, 2002,18(10): 490-491.
[24] Chen CX, Kwan CY. Endothelium-independent vasorelaxation by leonurine, a plant alkaloid purified from Chinese motherwort. Life Sci 2001, 68: 953-960.
[25] Wattanapitayakul SK, Bauer JA. Oxidative pathways in cardiovascular disease: roles, mechanisms, and therapeutic implications. Pharmacol Ther 2001, 89: 187-206.
[26] Ji XY, Tan BKH, Huang SH, Whiteman M, Zhu YC, Wu JY, Wei D, Zhu YZ. Effects of Salvia miltiorrhiza in ischemic myocardium in experimental rats. In:Perspective: Novel Compounds from Natural Products in New Millenniums. Tan B, Bay BH, Zhu YZ. eds. London: World Scientific Press, 2004,152-162.
[27] Zhu YZ, Huang SH, Tan BKH, Sun J, Whiteman M, Zhu YC. Antioxidants in Chinese herbal medicines: a biochemical perspective. Nat Prod Rep 2004, 21:478-489.
[28] Dhalla NS, Temsah RM, Netticadan T. Role of oxidative stress in cardiovascular diseases. J Hypertens, 2000, 18: 655-673.
[29] Yasmin W, Strynadka KD, Schulz R. Generation of peroxynitrite contributes to ischemia-reperfusion injury in isolated rat hearts. Cardiovasc Res, 1997, 33: 422-432.
[30] Rimm EB, Stampfer MJ. Antioxidants for vascular disease. Med Clin North Am, 2000, 84: 239-249.
[31] Zheng HA, Chen SR, Yin Jun. Effect of Leonurus Heterophyllus on oxygen free radicals during myocardial ischemia reperfusion injury in rabbits. J Shantou Univ Med Coll, 1997, 10(2): 10-12.
[32] Best PJ, Hasdai D, Sangiorgi G, Schwartz RS, Holmes DR Jr, Simari RD, Lerman A. Apoptosis-basic concepts and implications in coronary artery disease.Arterioscler Thromb Vase Biol, 1999,19: 14-22.
[33] Ferrari R, Agnoletti L, Comini L, Gaia G, Bachetti T, Cargnoni A, Ceconi C,Curello S, Visioli O. Oxidative stress during myocardial ischemia and heart failure. Eur Heart J, 1998,19:B2-B11.
[34] Turner NA, Xia F, Azhar G, Zhang XM, Liu LX. Oxidative stress induces DNA fragmentation and caspase activation via the JNK pathway in H9c2 cardiac muscle cells. J Mol Cell Cardiol, 1998, 30: 1789-1801.
[35] Hu X, Mao JC, Huang SH, Sun J, Wu YJ, Wei D, On T, Zhu YZ. Protective effects of Herba Leonuri in ischemic models. In: Natural Products: A Must for Human Survive. Zhu YZ, Tan B, Bay BH, Liu CH. eds. London: World Scientific Press, 2006, 361-379.
[36] Wang ZS, Li DW, Xia WJ, Qiu HQ, Zhu LY. The therapeutic effect of Herba Leonuri in the treatment of coronary myocardial ischemia. J Tradit Chin Med, 1988,8(2): 103-106.
[37] Chen HJ, Li DY, Saldeen T, Romeo F, Mehta JL. Mixed tocopherol preparation is superior to a-tocopherol alone against hypoxia-reoxygenation injury. Biochem Biophy Res Com, 2002, 291: 349-353.
[38] Loh KP, Low LS, Wong WH, Chou WH, Zhu YZ. Molecular mechanisms of cerebral protection from Ginkgo biloba extract on stroked rats. 2006; 398(1-2): 28-33.
[39] Hudlicka O, Brown M, Egginton S. Angiogenesis in skeletal and cardiac muscle. Physiol Rev, 1992, 72: 369-417.
[40] Schaper W. New paradigms for collateral vessel growth. Basic Res Cardiol, 1993, 88: 193-198.
[41] Waltenberger J. Modulation of growth factor action: implications for the treatment of cardiovascular diseases. Circulation, 1997, 96: 4083-4094.
[42] Li DW, Wang ZS, Qiu HQ. Herba Leonuri treatment for 80 patients with coronary heart diseases. Shanxi J Tradit Chin Med, 1984, 5 (10): 11-12.
[43] Zheng GW, Chen SR. Clinical observation of Herba Leonuri pill treatment for patients with myocardial ischemia. Chin J Inf Tradit Chin Med, 2000, 7(2): 19.
[44] Yan JZ, Chen SR. Curative effects of Leonuru Hererophyllus on coronary heart diseases. J Shantou Univ Med Coll, 2000, 13(1): 20-22.