艾炷灸循经取穴预处理对大鼠心肌缺血再灌注损伤心肌细胞凋亡基因的影响
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
实验目的:研究艾炷灸循经取穴预处理对大鼠心肌缺血再灌注损伤心肌细胞凋亡基因的影响。
     实验方法:通过艾炷灸循经取穴预处理2周,然后结扎大鼠左冠状动脉前降支(LAD),造成心肌缺血再灌注损伤模型,取心脏进行心肌病理学观察,免疫组化法检测Caspase-3;Bcl-2;Bax;Fas;TNF-α及NF-κB的表达。
     实验结果:(1)心肌缺血再灌注损伤后心肌细胞数量减少,细胞直径变小,细胞体积变小。艾炷灸(内关、郄门)预处理可以使心肌细胞数量增多,细胞直径变大,细胞体积变大。(2)凋亡标志因子Caspase-3在心肌缺血再灌注损伤后表达增多,艾炷灸(内关、郄门)预处理可降低其表达。(3)抑凋亡基因Bcl-2在心肌缺血再灌注损伤后表达减少,艾炷灸(内关、郄门)预处理可使Bcl-2表达增多。(4)促凋亡基因Bax,TNF-α,NF-κB与Fas在心肌缺血再灌注损伤后表达增高,艾炷灸(内关、郄门)预处理可使Bax,Fas,TNF-α与NF-κB表达降低。
     实验结论:艾炷灸循经取穴(内关、郄门)预处理对大鼠心肌缺血再灌注损伤心肌细胞凋亡起抑制作用。其作用可能是通过降低Caspase-3表达,增高Bcl-2表达,降低Bax,Fas,NF-κB与TNF-α表达来表现。
Objective:To study the effect of moxbustion on PC4 and PC6 on mycardial apoptosis induced by ischemia-reperfusion(I/R).
     Methods:The left anterior descene(LAD)coronary was ligate toestablish the ischemia-reperfusion(I/R)heart model in rats.Immunohistochemical staining was to detect the changes of apoptosis and protein expression of Caspase-3,Bcl-2,Bax,Fas, TNF-αand NF-κB.Histopathological changes in myocardium was also observed. Results:(1)The number and size of cardiomycyte cell reduced after ischemia-reperfusion and increased by moxbustion.(2)The expression of caspase-3 gene increased by ischemia-reperfusion and decreased by moxbustion.(3)The expression of Bcl-2 gene decreased by ischemia-reperfusion and increased by moxbustion.(4) The expression of Bax,Fas,TNF-αand NF-κB gene inregulating after ischemia-reperfusion and downregulating after moxbustion.
     Conclusions:Moxbustion on PC4 and PC6 had obviously preventive and therapeutic effects on reperfusion injury and could inhibit cardiomycyte apoptosis.The mechanism might be that Bcl-2 gene expression was increased,Caspase-3 gene,Bax gene,Fas gene,TNF-αgene and NF-κB gene were decreased following ischemia and reperfusion.
引文
1.国家“九五”科技攻关课题协作组.我国中年人群心血管病主要危险因素流行现状及从80年代初至90年代末的变化趋势.中华心血管病杂志,2001,29(2):74
    2.孔灵芝,慢性非传染性疾病的流行现状、发展趋势及防治策略,中国慢性病预防与控制,2002,10(1):1-2,19
    3.刘胜中,杨双强,心肌缺血/再灌注损伤机制研究进展,实用医院临床志,2007,4(1):88-90
    4.马国强,李跃荣,心肌缺血/再灌注损伤与细胞凋亡的机制及其防治研究进展,滨州医学院学报2007,30(1):50-52
    5.Yaoita H,Ogawa K,et al.Attenuation of ischemia reperfusion injury in rats by a caspase inhibitor.Circulation,1998,97(3):276)
    6.周仲英主编.中医内科学.北京.中国中医药出版社,2003,11
    7 黄春林,心血管科专病中医临床诊治,第2版,北京:人民卫生出版社,2005:76
    8 史载祥,后再灌注时代难题的中西医结合治疗思考,中西医结合心脑血管病杂志,2005,3(1):2
    9.Schulze CJ,Wang W,Kunari R,et al.Im balance between tissue inhibitor of metalloproteinase-4 and metalloproteinases during acute correction of myocardialischemia-reperfusioninjury[J]Circulation,2003,107(19):2487-2492
    10.李凯,郑世营,心肌缺血再灌注损伤与心肌细胞凋亡的研究进展,医学综述,2008,14(1):6-8
    11.Nepomniashchisk LM.Ultrastrutural mechanisms of myocardial atrophy in white rats during starvation.Biul Eksp BiolMed,1989,107:477-81
    12.Gottlieb RA et al.Reperfusion injury induces apoptosis in rabbits cardiomyocytes.J Clin Invest,1994:94(4):1621-83
    13.Kajstura J,Cheng W,Reiss K,etal.Apoptotic and necrotic myocyte cell death are independent contributing variables of infarct size in rats.Lab Invest,1996,74(1):86
    14.Tanaka M,Ito H,Adachi S,etal.Hypoxia induces apoptosis with enhanced expression of Fas antigen messenger RNA in culturedneonata rat cardiomyocyte.Circ Res,1994,75(3):426
    15.Chakrabarti S,HoqueAN,Karmazyn M.A rapid ischemia-induced apoptosis in isolated rat hearts and its attenuation by the sodium-hydrogen exchange inhibitor HOE 642(cariporide).J Mol cell Cardiol,1997,29(11):3169
    16.Gottlieb RA,Burleson KO,Kloner RA,etal.Reperfusion injury induces apoptosis in rabbit cardiomyocytes.J Clin Invest,1994,94(3):1621
    17.Maulik-N,Yoshida T,Engelman RM,etal.Ischemic preconditioning attenuates apoptotic cell death associated with ischemia/reperfusion.Mol Cell Biochem,1998,186(1-2):139
    18 FlissH,Gattinger D.Apoptosis in ischemia and reperfused rat myocardium.Circ Res,1996,79(5):949
    19.张峰,曹云新,罗晓星,等.缺氧诱导的心肌细胞凋亡及一氧化氮的保护作用.细胞与分子免疫学杂志,2000,16(3):225
    20.常志文,刘琦.缺血再灌注心肌细胞凋亡及药物干预.中国新药杂志,2000,9(1):28
    21.Fliss H et al.Apoptosis in ischemic and reperfused rat myocardium.Circ Res,1996;79:949-56
    22.SarasteA,etal.Apoptsis in human acute myocardial infarction,Circulation,1997,95:320
    23.Kajstura J,Cheng W,Reiss K,etal.Apoptotic and necrotic myocyte cell death are independent contributing variables of infarct size in rats.Lab Invest,1996,74(1):86
    24.Nicholson D W.Caspase structure,proteolytic substrates and function during apoptotic cell death[J].Cell Death Differ,1996,6(11):1028-1042
    25.Fan T J,Han LH,Cong RS,etal,Caspase family protease and apoptosis[J].Acta Biochim Biophys sin(Shanghai),2005,37(1):719-727
    26.RuiXing Y,Al-Ghaza li R,Wenwul,etal.Pretreatmennt with prbucol attenuates,cardiomyocyte apoptosis in a rabbit model of ischemia/reperfusion[J].Scand J clin Lab Invest,2006,66(7):549-558
    27.Vohra HA,Galinanes M.Effect of the degree of ischaemic injury and reoxyenation time on the type of myocardial cell deatf in man:role of caspase[J].BMC Physiol,2005,5(3):14
    28.徐强,司良毅,张红等,大鼠心肌再灌注不同时相caspase-3激活与心功能变化的关系,第三军医大学学报,2005,27(23):2338-2340
    29.姜明,曹士奇,陈锁成等,Caspase-3抑制剂对兔体外循环心肌细胞凋亡的影响,江苏大学学报(医学版),2007,17(3):205-208
    30.周艳玲,徐明,张玉芹等,大鼠急性心肌缺血再灌注心肌细胞凋亡与Bcl-2/Bax蛋白的表达,武警医学,2003,12(3):131-135
    31.徐军美,谭嵘,胡冬煦等,缺血预处理对兔缺血再灌注心肌bc-2,bax,p53基因表达的影响,湖南医科大学学报,2003,28(2):111-113
    32.郭靖,介曙光,曹向戎,心肌缺血再灌注损伤过程中细胞凋亡的实验研究,华北煤炭医学院学报,2006,8(6):758-760
    33.Jeremias J,Kupptt C,Martin-Villalba A,etal.Involvement of CD95/Apol/Fas in cell death after myocardial ischmia.Circulation,2000,102:915-920
    34.Zhao M,Chen Y,Li Y,Changes of apoptosis and Fas gene expression in cardiomycytes of rats with preconditioning.Zhonghua Nei Ke Za Zhi,1995,38(11):753-756
    35.Ding YF,Zhang MM.He RR.Ischemic preconditioning reduces cardiomyytic apoptosis in rabbit heart in vivo,Sheng Li Xue Bao,2000,52(3):220-224
    36.徐强,司良毅,张红,大鼠心肌再灌注不同时相Fas表达和半胱胺酸蛋白酶-3活性变化规律,中国临床康复,2004,8(27):5828-5830
    37.徐少东,马礼坤,屈朝法等,犬急性心肌梗死晚期再灌注对心肌Fas/FasL系统的影响及其可能的氧化应激机制,中国病理生理杂志,2007,23(2):307-310
    38.Sen R,Baltimore D.Multiple nuclear factors interact with the immunoglobulinenhancer sequences[J].Cell,1986,46(5):705-71
    39.Li C,Browder W,Kao RL.Early activation of transcription factorNF-κ B during ischemia in perfused rat heart[J].Am J Physiol,1999,276(2):543-552.
    40.Boyle EM,Canty TG,Morgan EN,et al.Treating myocardial Ischemia-reperfusion injury by targeting endothelial cell transcription[J].Ann Thorac Surg,1999,68(5):1949-1953.
    41.Canty TG,Boyle EM,Farr A,et al.Oxidative Stress induces NF-kappaB nuclear translocation without degradation of IkappaBalpha[J].Circulation,1999,100(Suppl 19):361-364.
    42.Maulik N,Goswami S,Galang N,et al.Differential regulation of Bcl-2,AP-1 and NF-kappaB on cardiomyocyte apoptosis during myocardial ischemic stress adaptation[J].FEBS Lett,1999,443(3):331-336.
    43.Frangogiannis NG,Lindsey ML,Michael LH.Resident Cardiac mast cells degranulate and release performed TNF-alpha,initing the cytokine cascade in experimental Canine myocadial ischemia/reperfusion[J].Circulation,1998;98(7):699-710.
    44.Dainel R,Meldrum.Tumor necrosis factor in the heart[J].AM J Physiol 1998,274(3pt2):R577-595.
    45.Haunstetter A,Izumo S,Apoptosis:dasic mechanisms and implications for cardiovascular disease[R].Circ Res,1998,82(11):1111
    46.唐旭尔,姜建青,刘宝玉等,心肌缺血再灌注中肿瘤坏死因子-α表达及多层次的心肌保护,四川医学,2004,25(2):129-132
    47.丁文惠,吴付轩,李大元等.肿瘤坏死因子在心肌缺血再灌注中的变化和意义[J].中国介入心脏病学杂志,1998,6(2):51-53
    48.Meldrum DR,Partrick DA,Cleveland JC,et al.On-pump coronary artery bypass surgery activates human myocardial NF-kappaB and increases TNF-alpha in the heart [3].J Surg Res,2003,112(2):175-179
    49刁利红,针刺内关穴为主治疗冠心病心绞痛临床观察,辽宁中医杂志,2003,30(8):667-668
    50.张朝晖,王强.针刺内关神门对冠心病患者血小板活性的影响[J].中国针灸,2000,20(2):119-120.
    51 于礼,王天佑.针刺内关治疗冠心病98例[J].山东中医杂志,1999,18(7):213.
    52.丛华,贾青顺,杨卫东.针刺内关穴对高压力负荷冠心病心绞痛患者心脏动力学的影响[J].山东中医杂志,2003,22(7):415-416.
    53.范爱新,朱华芝,裴广祥.针刺吸氧疗法在急性心肌梗死救治中的应用[J].中国针灸,1999,19(4):213.
    54.田岳凤,吴富东,乔海法等,针刺手厥阴心包经穴对心肌缺血再灌注损伤细胞凋亡的影响,中国针灸,2003,23(11):685-688
    55.田岳凤,吴富东,乔海法等,针刺手厥阴心包经穴对心肌缺血再灌注损伤凋亡调控基因表达的影响,中国中医基础医学杂志,2003,9(9):697-700
    56.田岳凤,针刺手厥阴心包经穴对心肌缺血再灌注损伤NO及NOS的影响.山东中医药大学学报,2003,27(2):150-151
    57.田岳凤,吴富东,乔海法等,针刺手厥阴心包经穴对心肌缺血再灌注损伤肌钙蛋白T 的影响,中国中西医结合急救杂志,2003,10(1):3-5
    58.林亚平,易受乡,严洁等,针刺内关对心肌缺血-再灌注损伤家兔CGRP和PGE2的影响,中国中医药信息杂志,2003,10(8):26-28
    59 高俊虹,付卫星,晋志高等.电针预治疗保护心肌缺血再灌注损伤-β肾上腺素受体的耐受机制,针刺研究,2006,31(1):22-26
    60.李春磊,孙忠人,宋春燕等,针刺预处理对心肌缺血大鼠血清S O D、M D A水平的影响,针灸临床杂志,2006,22(6):46-47
    61.严洁,田岳凤,刘健华等,电针内关对心肌缺血再灌注损伤N O及N O S的影响[J].中国中医基础学杂志,2002,8(7):68-70
    62.田岳凤,容受乡,严洁等,电针内关穴对心肌缺血-再灌注损伤预防保护作用的实验研究(Ⅰ)-对CK、ET、MD、NO、NOS等的影响[J].中国中西医结合急救杂志,2002,1(9):12-14
    63.方传龙.针刺”内关”穴区对I S P所致的心肌损伤的形态学研究[J].针灸临床杂志,2001,17(7):46-48
    64.王友京,董晓彤,王双昆等.电针对急性心肌缺血大鼠血浆中ET、NO和C G R P含量的影响[J].中国中医基础医学杂志,2002,8(6):74-75,80
    65.Yamasaki,Sato T,Sugimotok,et al.Effects of diltiazem on sympthetic hyperactivity in patients with vasospasic angina as assessed by spectral analysis of arterial pressure and heart rate variability[J]. Am J Cardiol, 1998, 81:137
    
    66. FuKazawa M, Adachi Y, Akima M, et al. Effects of CP-060S,a novel cardioprotective drug, in the methacholine-induced ECG change model in rats[J]. Fundam clin pharmacol. 2001,15:135
    
    67. Deinirag K, Askar FZ, Uyar M, et al. The protective effects of high does ascorbic acid diltiazem on myocardial ischaemia reperfusion injury[J]Middle East J Anesthesiol, 2001,16:67
    1.刁利红,针刺内关穴为主治疗冠心病心绞痛临床观察,辽宁中医杂志,2003,30(8):667-668
    3.王锐.辨证针刺治疗冠心病心绞痂42例[J].中国针灸,2003,23(5):280
    4 李保良,庞通.针刺背俞穴治疗冠心病心绞痛临床疗效观察[J].中国针灸,1999,(7):405-406.
    5 孟竞璧.针刺治疗冠心病的疗效及其作用机理研究[J].中医杂志,2002,43(8):587-589.
    6 张朝晖,王强.针刺内关神门对冠心病患者血小板活性的影响[J].中国针灸,2000,20(2):119-120.
    7 于礼,王天佑.针刺内关治疗冠心病98例[J].山东中医杂志,1999,18(7):213.
    8 李红霞,刘世伟,艾灸背俞穴治疗冠心病30例临床观察,职业与健康,2000,16(3):74
    9.齐惠涛,宋春霞,背俞穴隔姜灸治疗胸痹,针灸临床杂志,1998,14(1);35
    10 刘振义,刘勇,悬灸治疗冠心病62例临床研究,中医外治杂志,1996,3:6
    11 吴华慧,王晓丽,张霁.针药结合治疗冠心病心绞痛的临床观察[J].针灸临床杂志,2005,21(12):21
    12 赵艳玲,唐晨光,黄洁,刘剑勇,王群.针药并用治疗冠心病心绞痛临床疗效观察及对血浆ET、cGRP影响研究[J].中国针灸,2001,21(2):70-72
    13 袁中君,于淼.针药结合治疗冠心病心绞痛临床观察[J].辽宁中医学院学报,2002,4(4):306.
    14 李慧,庄玲玲,陶萍,等针刺结合丹参注射液治疗心绞痛36例[J].吉林中医药,2003,23(1):37.
    15.粱春雨,盛丽娟.针药结合治疗冠心病心绞痛46例临床观察[J].针刺研究,1998,(4):303-304.
    16 曹建萍,高峰,罗侃,等.针药结合治疗不稳定型心绞痛临床疗效及动态心电图观察[J].中国针灸,2002,22(6):363-365.
    17 韩艾.针刺在治疗冠心病患者过程中的抗氧自由基作用[J].中国针灸,1999,19(12):745.
    18 王兴,张友宝,姜波.针刺治疗不稳定型心绞痛的临床疗效观察[J].针灸临床杂志,2000,16(3):15.
    19 田岳凤,吴富东,乔海法,等针刺手厥阴心包经穴对心肌缺血再灌注损伤细胞凋亡的影响,中国针灸,2003,23(11):685
    20 田岳凤,吴富东,乔海法,等针刺手厥阴心包经穴对心肌缺血雨灌注损伤凋亡调控基因表达的影响,中国中医基础医学杂志,2003,9(9)57-58
    21 田岳凤,针刺手厥阴心包经穴对心肌缺血再灌注损伤NO及NOS的影响.山东中医药大学学报,2003,27(2):150-151
    22 田岳凤,吴富东,乔海法等,针刺手厥阴心包经穴对心肌缺血再灌注损伤肌钙蛋白T的影响,中国中西医结合急救杂志,2003,10(1):3-5
    23.林亚平,易受乡,严洁等,针刺内关对心肌缺血-再灌注损伤家兔CGRP和PGE2的影响,中国中医药信息杂志,2003,10(8):26-28
    24 高俊虹,付卫星,晋志高等.电针预治疗保护心肌缺血再灌注损伤-β肾上腺素受体的耐受 机制,针刺研究,2006,31(1):22-26
    25.李春磊,孙忠人,宋春燕等,针刺预处理对心肌缺血大鼠血清S O D、M D A水平的影响,针灸临床杂志,2006,22(6):46-47