甘氨酸对心肌缺血—再灌注损伤的防治作用及机制研究
|
摘要
目的：甘氨酸(glycine，GLY)是一种最简单的氨基酸。在我们先前的研究中发现，GLY不仅可以直接拮抗内毒素的多种生物学活性，而且还具有保护心脏的作用，能够有效地对抗内毒素所致的心肌损伤及垂体后叶素(pituitrin,Pit)诱导的心肌缺血性损伤。本实验是在上述研究的基础上，进一步探讨GLY对心肌缺血-再灌注损伤(myocardial ischemia-reperfusion,MI/R)的防治作用及机制，为防治缺血性心脏病提供新的思路与方法。
方法：小鼠随机分为5组，以不同药物定时定量灌胃(0.25mL／10g，2次／d)处理。1周后，腹腔注射Pit30U／kg，间隔30min后再腹腔注射硝酸甘油(nitroglycerin,Nit)10mg/kg复制急性MI／R模型。同时记录不同时点的肢体Ⅱ导联心电图，观察小鼠心电图J点及心率的变化。4h后，取出小鼠心脏，分别用比色法和硝酸还原酶法，检测各组小鼠心肌组织中一氧化氮合酶(nitri coxide synthase,NOS)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的活性及一氧化氮(nitric oxide,NO)的含量，观察GLY对这些活性物质的影响；用逆转录-多聚酶链式反应(RT-PCR)测定各组小鼠心肌组织Bcl-2 mRNA的表达，观察GLY对其表达量的影响。
结果：心电图结果显示，Pit诱导心肌缺血的动物，心电图J点明显降低，心率明显减慢。MI／R+Gly组在不同时点的J点及心率变化幅度较小，显著小于MI／R组(P＜0.05，P＜0.01)；MI／R+Gly组心肌组织总NOS活性及NO含量升高，而iNOS活性下降，与MI／R组相比，有显著性差异(P＜0.01)；MUR+Gly组Bcl-2扩增产物也较MI／R组明显增多(P＜0.01)。
结论：预防性口服适量GLY有利于保护心脏，减轻MI／R引发的损伤，维持良好的心功能状态。表现在心电图上，GLY降低急性心肌缺血动物心电图J点及心率的变化幅度。GLY的这一作用可能与其增加缺血再灌注心肌组织中NOS的活性及NO的生成，抑制iNOS的活性；促进缺血再灌注心肌Bcl-2 mRNA的表达有关。
Objective: Being the simplest amino acid, glycine not only acts as an anti-endotoxin agent, but also has great effects on cytoprotection. It has been demonstrated that glycine protected against myocardial injury induced by endotoxin and acute ischemia in our laboratory works. For contributing to the treatment for ischemic heart disease, the aim of this study was to investigate the protection of glycine against myocardial ischemia-reperfusion and its mechanism.
Methods: Mice were randomly divided into five groups and fed with different solution (0.25mL/10g, twice per day). On the 8th day, the mice were injected with pituitrin (30U/kg) and nitroglycerin (10mg/kg, 30 min later) intraperitoneally to remodel acute myocardial ischemia-reperfusion (MI/R). ECG and heart rate were recorded at Omin, 5min, lOmin, 20min, 30min, 40min, 50min. 4h later, the activities of nitric oxide synthase, inducible nitric oxide synthase, and concentrations of nitric oxide in myocardium were examined; The expression of Bcl-2 mRNA in myocardium was also determined by RT-PCR.
Results: J point of ECG and heart rate decreased obviously in MI/R-injured mice. Compared to Group MI/R, J point of ECG and heart
rate decreased smoothly in Group MI/R+GLY (P<0.05, P<0.01). The activities of NOS and concentrations of NO in Group MI/R+GLY were higher than those in Group MI/R (P<0.01), but the activities of iNOS was lower inversely (P<0.01). In addition, the expression of Bcl-2 mRNA in Group MI/R+GLY was higher than that in Group MI/R (P<0.01).
Conclusions: The results suggested that pretreatment of glycine attenuated myocardial ischemia-reperfusion injury induced by pituitrin and nitroglycerin. The mechanism may be related to NOS activation, the increase of NO content, the expression of Bcl-2 mRNA, and the reduction of iNOS in I/R-injured myocardium.
方法：小鼠随机分为5组，以不同药物定时定量灌胃(0.25mL／10g，2次／d)处理。1周后，腹腔注射Pit30U／kg，间隔30min后再腹腔注射硝酸甘油(nitroglycerin,Nit)10mg/kg复制急性MI／R模型。同时记录不同时点的肢体Ⅱ导联心电图，观察小鼠心电图J点及心率的变化。4h后，取出小鼠心脏，分别用比色法和硝酸还原酶法，检测各组小鼠心肌组织中一氧化氮合酶(nitri coxide synthase,NOS)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)的活性及一氧化氮(nitric oxide,NO)的含量，观察GLY对这些活性物质的影响；用逆转录-多聚酶链式反应(RT-PCR)测定各组小鼠心肌组织Bcl-2 mRNA的表达，观察GLY对其表达量的影响。
结果：心电图结果显示，Pit诱导心肌缺血的动物，心电图J点明显降低，心率明显减慢。MI／R+Gly组在不同时点的J点及心率变化幅度较小，显著小于MI／R组(P＜0.05，P＜0.01)；MI／R+Gly组心肌组织总NOS活性及NO含量升高，而iNOS活性下降，与MI／R组相比，有显著性差异(P＜0.01)；MUR+Gly组Bcl-2扩增产物也较MI／R组明显增多(P＜0.01)。
结论：预防性口服适量GLY有利于保护心脏，减轻MI／R引发的损伤，维持良好的心功能状态。表现在心电图上，GLY降低急性心肌缺血动物心电图J点及心率的变化幅度。GLY的这一作用可能与其增加缺血再灌注心肌组织中NOS的活性及NO的生成，抑制iNOS的活性；促进缺血再灌注心肌Bcl-2 mRNA的表达有关。
Objective: Being the simplest amino acid, glycine not only acts as an anti-endotoxin agent, but also has great effects on cytoprotection. It has been demonstrated that glycine protected against myocardial injury induced by endotoxin and acute ischemia in our laboratory works. For contributing to the treatment for ischemic heart disease, the aim of this study was to investigate the protection of glycine against myocardial ischemia-reperfusion and its mechanism.
Methods: Mice were randomly divided into five groups and fed with different solution (0.25mL/10g, twice per day). On the 8th day, the mice were injected with pituitrin (30U/kg) and nitroglycerin (10mg/kg, 30 min later) intraperitoneally to remodel acute myocardial ischemia-reperfusion (MI/R). ECG and heart rate were recorded at Omin, 5min, lOmin, 20min, 30min, 40min, 50min. 4h later, the activities of nitric oxide synthase, inducible nitric oxide synthase, and concentrations of nitric oxide in myocardium were examined; The expression of Bcl-2 mRNA in myocardium was also determined by RT-PCR.
Results: J point of ECG and heart rate decreased obviously in MI/R-injured mice. Compared to Group MI/R, J point of ECG and heart
rate decreased smoothly in Group MI/R+GLY (P<0.05, P<0.01). The activities of NOS and concentrations of NO in Group MI/R+GLY were higher than those in Group MI/R (P<0.01), but the activities of iNOS was lower inversely (P<0.01). In addition, the expression of Bcl-2 mRNA in Group MI/R+GLY was higher than that in Group MI/R (P<0.01).
Conclusions: The results suggested that pretreatment of glycine attenuated myocardial ischemia-reperfusion injury induced by pituitrin and nitroglycerin. The mechanism may be related to NOS activation, the increase of NO content, the expression of Bcl-2 mRNA, and the reduction of iNOS in I/R-injured myocardium.
引文
[1] 阮存珍，唐进，王子林．心肌缺血—再灌注损伤与药物预防．山西医药杂志 2001，30(6)：507-509
[2] 刘皋林，安登魁，张正行．一氧化氮与心血管疾病．药学进展1999，23(3)：136-142
[3] Jean-Luc Balligand, Paul J. Nitric Oxide Synthases and Cardiac Muscle. Arteriosclerosis, Thrombosis, and Vascular Biology 1997, 17. 1846-18
[4] Uwe Mehlhorn, Wilhelm Bloch, Andreas Krahwinkel, Karl LaRose, Hans-Joachim Geissler, Khosro Hekmat, Klaus Addicks, E. Rainer de Vivie. Activation of Myocardial constitutive nitric oxide synthase during coronary artery surgery. European Journal of Cardio-thoracic Surgery 2000, 17: 305-311
[5] Wenjie Wang, Grzegorz Sawicki, Richard Schulz. Peroxynitrite-induced myocardial injury is mediated through matrixmetalloproteinase-2. Cardiovascular Research, 2002, 53: 165-174
[6] 崔世涛，朱洪生．一氧化氮及其合酶在心肌缺血再灌注损伤中作用的研究．中华胸心血管外科杂志 1999，15 (6)：364-366
[7] Kanno S, Lee PC, Zhang Y, Hoc, Griffith BP, Shears LL, Billiar TR. Attenuation of myocardial ischemia / reperfusion injury by superinduction of inducible nitric oxide synthase. Circulation, 2000, 101 (23): 2742-2748
[8] 黄展勤，石刚刚．细胞凋亡与缺血性心脏病．汕头大学医学院学报．2001，14(2)：148-150
[9] Fliss H. Apoptosis in ischemia and reperfusioned rat myocardium. Circulation Research. 1996, 79: 949-956
[10] Zhi-Qing Zhao, Masanori Nakamura, Ning-Ping Wang, Josiah N. Wilcox, Steven Shearer, Russell S. Ronson, Robert A. Guyton, Jakob Vinten-Johansen. Reperfusion induces myocardial apoptotic cell death. Cardiovascular Research, 2000, 45: 651-660
[11] Peitan Liu, Baohuan Xu, Thomas A Cavalieri, Carl E. Hock. Age-related difference in myocardial function and inflammation in a rat model of myocardial ischemiareperfusion. Cardiovascular Research, 2002, 56:443-453
[12] Ulrike Weiland, Judith Haendeler, Christian Ihling. Inhibition of endogenous nitric oxide synthase potentiates ischemia-reperfusion-induced myocardial apoptosis via a caspase-3 dependent pathway. Cardiovascular Research, 2000, 45: 671-678
[13] Andrea Castro, Maria Cecilia Johnson, Mabel Anido, Armando Cortinez, Fernando Gabler, Margarita Vega. Role of nitric oxide and bcl-2 family genes in the regulation of human endometrial apoptosis. Fertility and Sterility, 2002, 78(3): 587-595
[14] Gao F, Gao E, Yue TL, Ohlstein EH, Lopez BL, Christopher TA, Ma XL. Nitric oxide mediates the antiapoptotic effect of insulin in myocardial ischemia-reperfusion: the roles of PI3-kinase, Akt, and endothelial nitric oxide synthase phosphorylation. Circulation, 2002, 105 (12): 1497-502.
[15] 陆大祥，李楚杰，付咏梅，王华东．甘氨酸对内毒素致热性的拮抗作用研究．中国病理生理杂志，1996，12(3)：235-237
[16] 孙葳，陆大祥，丁勇，李楚杰．不同pH及封闭氨基端对甘氨酸拮抗内毒素作用的影响．中国病理生理杂志，1999，15(2)：110-112
[17] 孙葳，陆大祥，丁勇，李楚杰．甘氨酸对内毒素与单核细胞结合率及内毒素构型的影响。中国病理生理杂志，1998，14(4)：359-361
[18] 丁勇，陆大祥，孙葳，张穗梅，李楚杰．甘氨酸对内毒素诱导单核细胞产生IL-1和TNF的影响．中国药理学通报，1999，15(2)：184-185
[19] 戚仁斌、陆大祥、王华东．甘氨酸对内毒素所致心肌损伤的影响．中国病理生理杂志，2001，17(8)：803-803．
[20] 周琼，陆大祥，付永梅，王华东．甘氨酸对小鼠心肌缺血性损伤的防治作用研究[J]．中国病理生理杂志，2002，18(4)：360-362
[21] Wheeler M, Stachlewitz RF, Yamashina S, Ikejima K, Morrow AL, Thurman RG. Glycine-gated chloride channels in neutrophils attenuate calcium influx and
superoxide production. FASEB journal, 2000, 14 (3): 476-484
[22] Yamashina S, Konno A, Wheeler MD, Rusyn I, Rusyn EV, Cox AD, Thurman RG. Endothelial cells contain a glycine-gated chloride channel. Nutrition and cancer, 2001, 40 (2): 197-204
[23] Li X, Bradford BU, Wheeler MD, Stimpson SA, Pink HM, Brodie TA, Schwab JH, Thurman RG. Dietary glycine prevents peptidoglycan polysaccharide-induced reactive arthritis in the rat: role for glycine-gated chloride channel. Infection and immunity, 2001, 69 (9): 5883-5891
[24] Edelstein CL, Ling H, Gengaro PE, Nemenoff RA, Bahr BA, Schrier RW. Effect of glycine on prelethal and postlethal increases in calpain activity in rat renal proximal tubules. Kidney international, 1997, 52 (5): 1271-1278
[25] Nichols JC, Bronk SF, Mellgren RL, Gores GJ. Inhibition of nonlysosomal calcium-dependent proteolysis by glycine during anoxic injury of rat hepatocytes. Gastroenterology, 1994, 106 (1): 168-176
[26] Wheeler MD, Thurman RG. Production of superoxide and TNF-alpha from alveolar macrophages is blunted by glycine. American journal of physiology, 1999, 277(5): L952-959
[27] Wheeler MD, Rose ML, Yamashima S, Enomoto N, Seabra V, Madren J, Thurman RG. Dietary glycine blunts lung inflammatory cell influx following acute endotoxin. Lung cellular and molecular physiology, 2000, 279 (2): L390-398
[28] Stachlewitz RF, Seabra V, Bradford B, Bradham CA, Rusyn I, Germolec D, Thurman RG. Glycine and uridine prevent D-galactosamine hepatotoxicitin the rat: role of Kupffer cells. Hepatology, 1999, 29 (3): 737-745
[29] Zhang Y, Ikejima K, Honda H, Kitamura T, Takei Y, Sato N. Glycine prevents apoptosis of rat sinusoidal endothelial cells caused by deprivation of vascular endothelial growth factor. Hepatology, 2000, 32 (3): 542-546
[30] M. D. Wheeler, K. Ikejema, N. Enomoto, R. F. Stacklewitz, V. Seabra, Z. Zhong, M. Yin, P. Schemmer, M. L. Rose, I. Rusyn, B. Bradford and R. G. Thurman.
Glycine: a new anti-inflammatory immuno-nutrient. Cellular and Molecular Life Sciences, 1999, 56:843-856
[31] Omasa M, Fukuse T, Toyokuni S, Mizutani Y, Yoshiha H, Ikeyama K, Hasegawa S, Wada H. Glycine ameliorates lung reperfusion injury after cold preservation in an ex vivo rat lung model. Transplantation, 2003, 75 (5): 591-598
[32] Yin M, Zhong Z, Connor HD, Bunzendahl H, Finn WF, Rusyn I, Li X, Raleigh JA, Mason RP, Thurman RG. Protective effect of glycine on renal injury induced by ischemia-reperfusion in vivo. Renal Physiology, 2002, 282(3): F417-423
[33] Lee MA, McCauley RD, Kong SE, Hall JC. Influence of glycine on intestinal ischemia-reperfusion injury. Journal of parenteral and enteral nutrition. 2002, 26(2): 130-135
[34] Ascher E, Hanson JN, Cheng W, Hingorani A, Scheinman M. Glycine preserves function and decreases necrosis in skeletal muscle undergoing ischemia and reperfu-sion injury. Surgery, 2001, 129(2): 231-235
[35] 汪长生，杨解人，桂常青，丁伯平，宋建国．丹参注射液对大鼠急性心肌缺血及血液流变学的影响．中国临床药理学与治疗学，2002，7(1)：30-32
[36] 张玲梅，夏洪生．抗心肌缺血再灌注损伤研究新进展．深圳中西医结合杂志，2001，11(1)：56-59
[37] 黄辉，吴伟康，陈建萍．新型缺血／再灌注模式诱导小鼠心肌细胞凋亡．中国心血管杂志．1999，4(3)：159-161
[38] 吴伟康，侯灿，吴金浪．垂体后叶素心肌缺血模型再探．中国病理生理杂志，1993，9(2)：124-126
[39] Watanabe T, Harumi K, Akustu Y, Yamanaka H, Okazaki O, Michihata T, Katagiri T. Significance of downsloping ST-segment depression induced by low-level exercise in severe coronary artery disease. Assessment with myocardial ischemia and collateral perfusion. Japanese Heart Journal, 1997, 38(2): 207-218
[40] 陈奇，陈长勋．心肌缺血和流量实验方法，中药药理实验方法．上海：上海科学技术出版社，1991，110-129
[41] 叶春玲，吕艳青，杨延秀，吕俊华，钟玲，任先达，刘声波．护心灵对垂体后叶素所致大鼠急性心肌缺血的保护作用．中成药，2001，23(11)：812-814
[42] 朱洪生，黄忠耀，崔世涛，张有荣，卢蓉，吴学军，张谷兰．实验性缺血再灌注心肌顿抑与一氧化氮的关系及东莨菪碱对其影响．中华心血管病杂志，1998，26(1)：65-67
[43] Tae-Jin Youn, Hyo-Soo Kim, Hyun-Jae Kang, Udo Albus, Wolfgang Scholz, Hartmut Ruetten, Andreas M. Zeiher, Stefanie Dimmeler. Inhibition of nitric oxide synthesis increases apoptotic cardiomyocyte death and myocardial angiotensin-converting enzyme gene expression in ischemia/reperfusion-injured myocardium ofrats. Heart Vessels 2001,16:12-19
[44] Masatora Iwashina, Masayoshi Shichiri, Fumiaki Marumo, Yukio Hirata. Transfection of inducible nitric oxide synthase gene causes apoptosis in vascular smooth muscle cells. Circulation, 1998, 98:1212-1218
[45] Friedrich Brunner, Robert Maier, Penelope Andrew, et al. Attenuation of myocardial ischemia/ reperfusion injury in mice with myocyte-specific overexpression of endothelial nitric oxide synthase. Cardiovascular Research, 2003, 57:55-62
[46] Stephen M Wildhirt, Susanne Weismueller, Costas Schulze, Nicole Conrad, Arno Kornberg, Bruno Reichart. Inducible nitric oxide synthase activation after ischemia/ reperfusioncontributes to myocardial dysfunction and extent of infarct size in rabbits:evidence for a late phase of nitric oxide-mediated reperfusion injury. Cardiovascular Research, 1999, 43:698-711
[47] Wei Song, Xiangru Lu, Qingping Feng. Tumor necrosis factor-a induces apoptosis via inducible nitric oxide synthase in neonatal mouse cardiomyocytes. Cardiovascular Research, 2000, 45:595-602
[48] Margaret Arstall, Douglas Sawyer, Ryuji Fukazawa, Ralph A Kelly. Cytokine-mediated apoptosis in cardiac myocytesthe role of inducible nitric oxide synthase induction and peroxynitrite generation. Circulation Research, 1999, 85:829-840
[49] Verena Stangl, Gert Baumann, Karl Stangl, Stephan B. Felix. Negative inotropic mediators released from the heart after myocardial ischaemia-reperfusion. Cardiovascular Research, 2002, 53: 12-30
[50] 徐明，曾庆云，丁成萦．大鼠急性心肌缺血再灌注的心肌细胞凋亡研究．解剖学报，2002，33(2)：180-184
[51] Read JC. Bcl-2 and the regulation of programmed cell death. J Cell Biol, 1994, 124(1-2): 1-6
[52] Roberta A, Roberta G, Engl ER. Apoptosis in myocardial ischemia reperfusion. Annal New York Academic of Sciences, 1998,874: 412-426
[53] Yang J, Lix X, Bhalla K. Prevention of apoptosis by Bcl-2 release of cytochrome C from mitochondria block. Science, 1997, 275(5303): 1129-1132
[54] Maulik N, Engelman RM, Rousou JA, Flack JE, Deaton D, Das DK. Ischemic preconditioning reduces apoptosis by upregulating anti-death gene Bcl-2. Circulation, 1999, 100 (19): Ⅱ369-375
[55] Yang L, Zhang B, Toku K, Maeda N, Sakanaka M, Tanaka J. Improvement of the viability of cultured rat neurons by the non-essential amino acids L-serine and glycine that upregulates expression of the anti-apoptotic gene product Bcl-w. Neuroscience letters, 2000, 295 (3): 97-100
[2] 刘皋林，安登魁，张正行．一氧化氮与心血管疾病．药学进展1999，23(3)：136-142
[3] Jean-Luc Balligand, Paul J. Nitric Oxide Synthases and Cardiac Muscle. Arteriosclerosis, Thrombosis, and Vascular Biology 1997, 17. 1846-18
[4] Uwe Mehlhorn, Wilhelm Bloch, Andreas Krahwinkel, Karl LaRose, Hans-Joachim Geissler, Khosro Hekmat, Klaus Addicks, E. Rainer de Vivie. Activation of Myocardial constitutive nitric oxide synthase during coronary artery surgery. European Journal of Cardio-thoracic Surgery 2000, 17: 305-311
[5] Wenjie Wang, Grzegorz Sawicki, Richard Schulz. Peroxynitrite-induced myocardial injury is mediated through matrixmetalloproteinase-2. Cardiovascular Research, 2002, 53: 165-174
[6] 崔世涛，朱洪生．一氧化氮及其合酶在心肌缺血再灌注损伤中作用的研究．中华胸心血管外科杂志 1999，15 (6)：364-366
[7] Kanno S, Lee PC, Zhang Y, Hoc, Griffith BP, Shears LL, Billiar TR. Attenuation of myocardial ischemia / reperfusion injury by superinduction of inducible nitric oxide synthase. Circulation, 2000, 101 (23): 2742-2748
[8] 黄展勤，石刚刚．细胞凋亡与缺血性心脏病．汕头大学医学院学报．2001，14(2)：148-150
[9] Fliss H. Apoptosis in ischemia and reperfusioned rat myocardium. Circulation Research. 1996, 79: 949-956
[10] Zhi-Qing Zhao, Masanori Nakamura, Ning-Ping Wang, Josiah N. Wilcox, Steven Shearer, Russell S. Ronson, Robert A. Guyton, Jakob Vinten-Johansen. Reperfusion induces myocardial apoptotic cell death. Cardiovascular Research, 2000, 45: 651-660
[11] Peitan Liu, Baohuan Xu, Thomas A Cavalieri, Carl E. Hock. Age-related difference in myocardial function and inflammation in a rat model of myocardial ischemiareperfusion. Cardiovascular Research, 2002, 56:443-453
[12] Ulrike Weiland, Judith Haendeler, Christian Ihling. Inhibition of endogenous nitric oxide synthase potentiates ischemia-reperfusion-induced myocardial apoptosis via a caspase-3 dependent pathway. Cardiovascular Research, 2000, 45: 671-678
[13] Andrea Castro, Maria Cecilia Johnson, Mabel Anido, Armando Cortinez, Fernando Gabler, Margarita Vega. Role of nitric oxide and bcl-2 family genes in the regulation of human endometrial apoptosis. Fertility and Sterility, 2002, 78(3): 587-595
[14] Gao F, Gao E, Yue TL, Ohlstein EH, Lopez BL, Christopher TA, Ma XL. Nitric oxide mediates the antiapoptotic effect of insulin in myocardial ischemia-reperfusion: the roles of PI3-kinase, Akt, and endothelial nitric oxide synthase phosphorylation. Circulation, 2002, 105 (12): 1497-502.
[15] 陆大祥，李楚杰，付咏梅，王华东．甘氨酸对内毒素致热性的拮抗作用研究．中国病理生理杂志，1996，12(3)：235-237
[16] 孙葳，陆大祥，丁勇，李楚杰．不同pH及封闭氨基端对甘氨酸拮抗内毒素作用的影响．中国病理生理杂志，1999，15(2)：110-112
[17] 孙葳，陆大祥，丁勇，李楚杰．甘氨酸对内毒素与单核细胞结合率及内毒素构型的影响。中国病理生理杂志，1998，14(4)：359-361
[18] 丁勇，陆大祥，孙葳，张穗梅，李楚杰．甘氨酸对内毒素诱导单核细胞产生IL-1和TNF的影响．中国药理学通报，1999，15(2)：184-185
[19] 戚仁斌、陆大祥、王华东．甘氨酸对内毒素所致心肌损伤的影响．中国病理生理杂志，2001，17(8)：803-803．
[20] 周琼，陆大祥，付永梅，王华东．甘氨酸对小鼠心肌缺血性损伤的防治作用研究[J]．中国病理生理杂志，2002，18(4)：360-362
[21] Wheeler M, Stachlewitz RF, Yamashina S, Ikejima K, Morrow AL, Thurman RG. Glycine-gated chloride channels in neutrophils attenuate calcium influx and
superoxide production. FASEB journal, 2000, 14 (3): 476-484
[22] Yamashina S, Konno A, Wheeler MD, Rusyn I, Rusyn EV, Cox AD, Thurman RG. Endothelial cells contain a glycine-gated chloride channel. Nutrition and cancer, 2001, 40 (2): 197-204
[23] Li X, Bradford BU, Wheeler MD, Stimpson SA, Pink HM, Brodie TA, Schwab JH, Thurman RG. Dietary glycine prevents peptidoglycan polysaccharide-induced reactive arthritis in the rat: role for glycine-gated chloride channel. Infection and immunity, 2001, 69 (9): 5883-5891
[24] Edelstein CL, Ling H, Gengaro PE, Nemenoff RA, Bahr BA, Schrier RW. Effect of glycine on prelethal and postlethal increases in calpain activity in rat renal proximal tubules. Kidney international, 1997, 52 (5): 1271-1278
[25] Nichols JC, Bronk SF, Mellgren RL, Gores GJ. Inhibition of nonlysosomal calcium-dependent proteolysis by glycine during anoxic injury of rat hepatocytes. Gastroenterology, 1994, 106 (1): 168-176
[26] Wheeler MD, Thurman RG. Production of superoxide and TNF-alpha from alveolar macrophages is blunted by glycine. American journal of physiology, 1999, 277(5): L952-959
[27] Wheeler MD, Rose ML, Yamashima S, Enomoto N, Seabra V, Madren J, Thurman RG. Dietary glycine blunts lung inflammatory cell influx following acute endotoxin. Lung cellular and molecular physiology, 2000, 279 (2): L390-398
[28] Stachlewitz RF, Seabra V, Bradford B, Bradham CA, Rusyn I, Germolec D, Thurman RG. Glycine and uridine prevent D-galactosamine hepatotoxicitin the rat: role of Kupffer cells. Hepatology, 1999, 29 (3): 737-745
[29] Zhang Y, Ikejima K, Honda H, Kitamura T, Takei Y, Sato N. Glycine prevents apoptosis of rat sinusoidal endothelial cells caused by deprivation of vascular endothelial growth factor. Hepatology, 2000, 32 (3): 542-546
[30] M. D. Wheeler, K. Ikejema, N. Enomoto, R. F. Stacklewitz, V. Seabra, Z. Zhong, M. Yin, P. Schemmer, M. L. Rose, I. Rusyn, B. Bradford and R. G. Thurman.
Glycine: a new anti-inflammatory immuno-nutrient. Cellular and Molecular Life Sciences, 1999, 56:843-856
[31] Omasa M, Fukuse T, Toyokuni S, Mizutani Y, Yoshiha H, Ikeyama K, Hasegawa S, Wada H. Glycine ameliorates lung reperfusion injury after cold preservation in an ex vivo rat lung model. Transplantation, 2003, 75 (5): 591-598
[32] Yin M, Zhong Z, Connor HD, Bunzendahl H, Finn WF, Rusyn I, Li X, Raleigh JA, Mason RP, Thurman RG. Protective effect of glycine on renal injury induced by ischemia-reperfusion in vivo. Renal Physiology, 2002, 282(3): F417-423
[33] Lee MA, McCauley RD, Kong SE, Hall JC. Influence of glycine on intestinal ischemia-reperfusion injury. Journal of parenteral and enteral nutrition. 2002, 26(2): 130-135
[34] Ascher E, Hanson JN, Cheng W, Hingorani A, Scheinman M. Glycine preserves function and decreases necrosis in skeletal muscle undergoing ischemia and reperfu-sion injury. Surgery, 2001, 129(2): 231-235
[35] 汪长生，杨解人，桂常青，丁伯平，宋建国．丹参注射液对大鼠急性心肌缺血及血液流变学的影响．中国临床药理学与治疗学，2002，7(1)：30-32
[36] 张玲梅，夏洪生．抗心肌缺血再灌注损伤研究新进展．深圳中西医结合杂志，2001，11(1)：56-59
[37] 黄辉，吴伟康，陈建萍．新型缺血／再灌注模式诱导小鼠心肌细胞凋亡．中国心血管杂志．1999，4(3)：159-161
[38] 吴伟康，侯灿，吴金浪．垂体后叶素心肌缺血模型再探．中国病理生理杂志，1993，9(2)：124-126
[39] Watanabe T, Harumi K, Akustu Y, Yamanaka H, Okazaki O, Michihata T, Katagiri T. Significance of downsloping ST-segment depression induced by low-level exercise in severe coronary artery disease. Assessment with myocardial ischemia and collateral perfusion. Japanese Heart Journal, 1997, 38(2): 207-218
[40] 陈奇，陈长勋．心肌缺血和流量实验方法，中药药理实验方法．上海：上海科学技术出版社，1991，110-129
[41] 叶春玲，吕艳青，杨延秀，吕俊华，钟玲，任先达，刘声波．护心灵对垂体后叶素所致大鼠急性心肌缺血的保护作用．中成药，2001，23(11)：812-814
[42] 朱洪生，黄忠耀，崔世涛，张有荣，卢蓉，吴学军，张谷兰．实验性缺血再灌注心肌顿抑与一氧化氮的关系及东莨菪碱对其影响．中华心血管病杂志，1998，26(1)：65-67
[43] Tae-Jin Youn, Hyo-Soo Kim, Hyun-Jae Kang, Udo Albus, Wolfgang Scholz, Hartmut Ruetten, Andreas M. Zeiher, Stefanie Dimmeler. Inhibition of nitric oxide synthesis increases apoptotic cardiomyocyte death and myocardial angiotensin-converting enzyme gene expression in ischemia/reperfusion-injured myocardium ofrats. Heart Vessels 2001,16:12-19
[44] Masatora Iwashina, Masayoshi Shichiri, Fumiaki Marumo, Yukio Hirata. Transfection of inducible nitric oxide synthase gene causes apoptosis in vascular smooth muscle cells. Circulation, 1998, 98:1212-1218
[45] Friedrich Brunner, Robert Maier, Penelope Andrew, et al. Attenuation of myocardial ischemia/ reperfusion injury in mice with myocyte-specific overexpression of endothelial nitric oxide synthase. Cardiovascular Research, 2003, 57:55-62
[46] Stephen M Wildhirt, Susanne Weismueller, Costas Schulze, Nicole Conrad, Arno Kornberg, Bruno Reichart. Inducible nitric oxide synthase activation after ischemia/ reperfusioncontributes to myocardial dysfunction and extent of infarct size in rabbits:evidence for a late phase of nitric oxide-mediated reperfusion injury. Cardiovascular Research, 1999, 43:698-711
[47] Wei Song, Xiangru Lu, Qingping Feng. Tumor necrosis factor-a induces apoptosis via inducible nitric oxide synthase in neonatal mouse cardiomyocytes. Cardiovascular Research, 2000, 45:595-602
[48] Margaret Arstall, Douglas Sawyer, Ryuji Fukazawa, Ralph A Kelly. Cytokine-mediated apoptosis in cardiac myocytesthe role of inducible nitric oxide synthase induction and peroxynitrite generation. Circulation Research, 1999, 85:829-840
[49] Verena Stangl, Gert Baumann, Karl Stangl, Stephan B. Felix. Negative inotropic mediators released from the heart after myocardial ischaemia-reperfusion. Cardiovascular Research, 2002, 53: 12-30
[50] 徐明，曾庆云，丁成萦．大鼠急性心肌缺血再灌注的心肌细胞凋亡研究．解剖学报，2002，33(2)：180-184
[51] Read JC. Bcl-2 and the regulation of programmed cell death. J Cell Biol, 1994, 124(1-2): 1-6
[52] Roberta A, Roberta G, Engl ER. Apoptosis in myocardial ischemia reperfusion. Annal New York Academic of Sciences, 1998,874: 412-426
[53] Yang J, Lix X, Bhalla K. Prevention of apoptosis by Bcl-2 release of cytochrome C from mitochondria block. Science, 1997, 275(5303): 1129-1132
[54] Maulik N, Engelman RM, Rousou JA, Flack JE, Deaton D, Das DK. Ischemic preconditioning reduces apoptosis by upregulating anti-death gene Bcl-2. Circulation, 1999, 100 (19): Ⅱ369-375
[55] Yang L, Zhang B, Toku K, Maeda N, Sakanaka M, Tanaka J. Improvement of the viability of cultured rat neurons by the non-essential amino acids L-serine and glycine that upregulates expression of the anti-apoptotic gene product Bcl-w. Neuroscience letters, 2000, 295 (3): 97-100