标本配穴电针干预对慢性心肌缺血模型大鼠凋亡相关蛋白表达与线粒体超微结构的影响
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摘要
目的:观察标本配穴电针治疗对慢性心肌缺血模型大鼠凋亡相关蛋白表达及线粒体超微结构的效应及机制。方法:将70只SPF级Wistar雄性大鼠根据随机数字表法分为7组。采用盐酸异丙肾上腺素(ISO)注射制作慢性心肌缺血模型,LY294002组和IGF-1组分别加注LY294002和IGF-1溶液,所有针刺组在造模前进行电针针刺预处理。最后检测凋亡相关蛋白Caspase-3、Caspase-9、Bax、Bcl-2的表达及心肌线粒体超微结构的变化。结果:各组大鼠心肌组织内凋亡相关蛋白表达相比正常组均有不同程度升高(P<0.01),且心肌超微结构有不同程度损伤。与模型组比较,标本配穴组Caspase-3、Caspase-9、Bax均有明显下降(P<0.01),Bcl-2蛋白表达升高(P<0.01)。经电针治疗后,标本配穴组与IGF-1组心肌线粒体损伤程度一致明显改善,且优于内关组。结论:"标本配穴"电针治疗可调节凋亡相关蛋白的表达,减轻心肌线粒体结构损伤,其调节过程及机制有可能通过PI3K/Akt信号通路来实现其保护心肌细胞的腧穴协同作用。
Objective: To observe the effects and explore the mechanism of the combined Biao and Ben acupoints treatment on apoptosis-related protein expression and mitochondrial ultrastructure in rats with chronic myocardial ischemia.Methods: According to the random number table method, 70 normal Wistar male rats were divided into 7 groups. The chronic myocardial ischemia model was made by ISO injection, and the LY294002 group and IGF-1 group were respectively added to give LY294002 solution and IGF-1 solution injection. All acupuncture groups adopt electroacupuncture pretreatment before modeling.Finally, the expressions of apoptosis-related proteins Caspase-3, Caspase-9, Bax and Bcl-2 were tested and the changes of myocardial mitochondrial ultrastructure were observed. Results: The expression of apoptosis-related proteins in myocardial tissue of each group was increased in different degrees compared with the normal group(P<0.01), and the myocardial ultrastructure had different degrees of damage. Compared with the model group, the Caspase-3, Caspase-9 and Bax in the combined Biao and Ben acupoints treatment group were significantly decreased(P<0.01), and the Bcl-2 protein expression was increased(P<0.01).After electroacupuncture treatment, the degree of myocardial mitochondrial damage was significantly improved in the combined Biao and Ben acupoints treatment group and the IGF-1 group, and it was better than the Neiguan(PC6) group. Conclusion: The combined Biao and Ben acupoints treatment can regulate the expression of apoptosis-related proteins and alleviate myocardial mitochondrial structural damage, and the regulation process and mechanism may be through PI3 K/Akt signaling pathway to achieve the synergy of acupoints to protect cardiomyocytes.
Objective: To observe the effects and explore the mechanism of the combined Biao and Ben acupoints treatment on apoptosis-related protein expression and mitochondrial ultrastructure in rats with chronic myocardial ischemia.Methods: According to the random number table method, 70 normal Wistar male rats were divided into 7 groups. The chronic myocardial ischemia model was made by ISO injection, and the LY294002 group and IGF-1 group were respectively added to give LY294002 solution and IGF-1 solution injection. All acupuncture groups adopt electroacupuncture pretreatment before modeling.Finally, the expressions of apoptosis-related proteins Caspase-3, Caspase-9, Bax and Bcl-2 were tested and the changes of myocardial mitochondrial ultrastructure were observed. Results: The expression of apoptosis-related proteins in myocardial tissue of each group was increased in different degrees compared with the normal group(P<0.01), and the myocardial ultrastructure had different degrees of damage. Compared with the model group, the Caspase-3, Caspase-9 and Bax in the combined Biao and Ben acupoints treatment group were significantly decreased(P<0.01), and the Bcl-2 protein expression was increased(P<0.01).After electroacupuncture treatment, the degree of myocardial mitochondrial damage was significantly improved in the combined Biao and Ben acupoints treatment group and the IGF-1 group, and it was better than the Neiguan(PC6) group. Conclusion: The combined Biao and Ben acupoints treatment can regulate the expression of apoptosis-related proteins and alleviate myocardial mitochondrial structural damage, and the regulation process and mechanism may be through PI3 K/Akt signaling pathway to achieve the synergy of acupoints to protect cardiomyocytes.
引文
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[11]唐民科,张均田.半胱氨酸-天冬氨酸蛋白酶(Caspase)及其在细胞凋亡中的作用.医学研究杂志,2000,29(11):9-13
[12]Zhao Z Q,Velez D A,Wang N P,et al.Progressively developedmyocardial apoptotic cell death during late phase of reperfusion.Apoptosis,2001,6(4):279-290
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[15]李辉,石涵,朱天民,等.红豆杉多糖对犬心肌缺血再灌注损伤模型心肌组织损伤及超微结构的影响.中华中医药杂志,2012,27(1):219-223
[16]王晶波,吕铭洋,崔新明等.急性心肌缺血犬心肌超微结构的变化及龙牙葱木皂苷的保护作用.中国老年学杂志,2012,8(32):3460-3461
[17]田岳凤,王荣,李雷勇,等.针刺手厥阴经穴对心肌缺血/再灌注损伤线粒体超微结构的影响.针刺研究,2006,10(3):268-271
[18]周晓莉,刘品莉,姜青松,等.“双固一通”针法治疗绝经后骨质疏松症的临床研究.中国骨质疏松杂志,2008,14(3):204-207
[19]鲍圣涌,王华,王彦春,等.“双固一通”针法对绝经后骨质疏松症模型大鼠骨组织骨保护蛋白mRNA表达水平的影响.针刺研究,2006,31(3):119-152,178
[20]徐芬,梁凤霞,陈瑞,等.标本配穴电针对胰岛素抵抗大鼠胰岛素敏感性及骨骼肌SIRT1蛋白表达的影响.中医杂志,2014,4:328-332
[21]周焕娇,王静芝,刘建民,等.“标本配穴”电针对胰岛素抵抗大鼠下丘脑磷酸化糖原合成酶激酶3β表达的影响.湖北中医药大学学报,2014,16(1):20-23
[22]李强,徐芬,梁凤霞,等.“标本配穴”电针对胰岛素抵抗大鼠股四头肌线粒体超微结构和生物合成功能的影响.中国针灸,2014,34(6):578-582
[23]梁凤霞,徐芬,陈瑞,等.“标本配穴”针刺对糖尿病大鼠下丘脑瘦素受体及其mRNA表达的影响.中华中医药学刊,2012,30(12):2773-2775
[24]洪亚群,王华.“双固一通”针法对甲亢性心肌病大鼠AngⅡ、ET及CGRP的影响.中国康复,2008,23(3):150-151
[25]洪亚群,王华.“双固一通”针法对甲亢性心肌病大鼠肥厚心肌病理组织学影响的研究.世界中西医结合杂志,2008,3(6):319-321
[26]王彦春,马骏,王华.“双固一通”针法对帕金森病模型大鼠神经干细胞增殖及分化的影响.中国针灸,2006,26(4):277-279
[2]A K Jonassen,B K Brar,O D Mjos,et al.Insulin administeredat reoxygenation exerts a cardioprotective effect in myocytesby a Possible anti-apoptotic mechanism.J Mol Cell Cardiol,2000,32(5):757-764
[3]张清友,魏冰,宫丽敏,等.果糖二磷酸钠对大鼠心肌缺血的防治作用及其机制.中国药理学通报,2002,18(6):631-634
[4]林文注,王佩.实验针灸学.上海:上海科学技术出版社,1999:288
[5]王艳杰,邓雯,张鹏飞.细胞色素C与细胞凋亡研究进展.动物医学进展,2012,33(7):89-92
[6]朱汉华,李浪,汪熠,等.大鼠冠状动脉微栓塞致心肌细胞凋亡与天冬氨酸特异性半胱氨酸蛋白酶活化的动态变化及关系.中华老年医学杂志,2010,29(9):755-760
[7]S Faubel,C L Edelstein.Caspases as drug targets in ischemicorgan injury.Curr Drug Targets Immune Endocr MetabolDisord,2005,5(3):269-287
[8]Lockshin R A.Programmed cell death:history and future of aConcept.J Soc Biol,2005,199(3):169-173
[9]D Garcia-Dorado,P Théroux,J M Duran,et al.Selective inhibition ofthe contractile apparatus:A new approach to modification of infarctsize,infarct composition,and infarct geometry during coronary arteryocclusion and reperfusion.Circulation,1992,85(3):1160-1174
[10]Kai Sun,Zhi-Su Liu,Quan Sun,et al.Role of mitochondria in cellapoptosis during hepatic ischemia-reperfusion injury and protectiveeffect of ischemic postconditioning.World J Gastroenterol,2004,10(13):1934-1938
[11]唐民科,张均田.半胱氨酸-天冬氨酸蛋白酶(Caspase)及其在细胞凋亡中的作用.医学研究杂志,2000,29(11):9-13
[12]Zhao Z Q,Velez D A,Wang N P,et al.Progressively developedmyocardial apoptotic cell death during late phase of reperfusion.Apoptosis,2001,6(4):279-290
[13]R Aikawa,I Komuro,T Yamazaki,et al.Oxidative stressactivatesextracellular signal-regulated kinasesthroughSrcandRasincultured cardiac myocytesof neonatalrats.J Clin Inves,1997,100(7):1813-1821
[14]Olivettl G,Abbi R,Quaini F,et al.Apoptosis in the failing humanheart.N Engl J Med,1997,336(16):1311-1341
[15]李辉,石涵,朱天民,等.红豆杉多糖对犬心肌缺血再灌注损伤模型心肌组织损伤及超微结构的影响.中华中医药杂志,2012,27(1):219-223
[16]王晶波,吕铭洋,崔新明等.急性心肌缺血犬心肌超微结构的变化及龙牙葱木皂苷的保护作用.中国老年学杂志,2012,8(32):3460-3461
[17]田岳凤,王荣,李雷勇,等.针刺手厥阴经穴对心肌缺血/再灌注损伤线粒体超微结构的影响.针刺研究,2006,10(3):268-271
[18]周晓莉,刘品莉,姜青松,等.“双固一通”针法治疗绝经后骨质疏松症的临床研究.中国骨质疏松杂志,2008,14(3):204-207
[19]鲍圣涌,王华,王彦春,等.“双固一通”针法对绝经后骨质疏松症模型大鼠骨组织骨保护蛋白mRNA表达水平的影响.针刺研究,2006,31(3):119-152,178
[20]徐芬,梁凤霞,陈瑞,等.标本配穴电针对胰岛素抵抗大鼠胰岛素敏感性及骨骼肌SIRT1蛋白表达的影响.中医杂志,2014,4:328-332
[21]周焕娇,王静芝,刘建民,等.“标本配穴”电针对胰岛素抵抗大鼠下丘脑磷酸化糖原合成酶激酶3β表达的影响.湖北中医药大学学报,2014,16(1):20-23
[22]李强,徐芬,梁凤霞,等.“标本配穴”电针对胰岛素抵抗大鼠股四头肌线粒体超微结构和生物合成功能的影响.中国针灸,2014,34(6):578-582
[23]梁凤霞,徐芬,陈瑞,等.“标本配穴”针刺对糖尿病大鼠下丘脑瘦素受体及其mRNA表达的影响.中华中医药学刊,2012,30(12):2773-2775
[24]洪亚群,王华.“双固一通”针法对甲亢性心肌病大鼠AngⅡ、ET及CGRP的影响.中国康复,2008,23(3):150-151
[25]洪亚群,王华.“双固一通”针法对甲亢性心肌病大鼠肥厚心肌病理组织学影响的研究.世界中西医结合杂志,2008,3(6):319-321
[26]王彦春,马骏,王华.“双固一通”针法对帕金森病模型大鼠神经干细胞增殖及分化的影响.中国针灸,2006,26(4):277-279
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