电针预处理对心肌缺血再灌注无复流大鼠NF-kB p65蛋白及其磷酸化水平的影响
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摘要
目的观察电针预处理对大鼠心肌缺血再灌注无复流现象及核转录因子-?B(NF-?B) p65蛋白及其磷酸化水平的影响。方法 48只雄性SD大鼠随机分为假手术组、模型组、电针预处理组、阿托伐他汀预处理组。除假手术组外,其余各组大鼠均采用结扎左冠状动脉45 min后再灌注120 min的方法复制急性心肌缺血再灌注无复流的模型。观察各组大鼠心肌梗死、缺血及无复流的范围,酶联免疫吸附法(ELISA)测定血清炎性因子白细胞介素-10(IL-10)、白细胞介素-6(IL-6)和肿瘤坏死因子(TNF-?)的水平,蛋白质免疫印迹法(Western-blot)测定心肌组织中NF-?B p65及其磷酸化水平的表达。结果与模型组比较,电针预处理组和阿托伐他汀预处理组心肌梗死范围及无复流范围明显减小,电针预处理能显著升高心肌梗死大鼠血清IL-10水平,降低血清IL-6及TNF-?水平,下调心肌组织中NF-?B p65及phospho-NF-?B p65蛋白的表达,差异均有统计学意义(P<0.01,P<0.05)。结论电针预处理能够显著降低心肌组织中NF-?B p65及其磷酸化水平的表达,从而改善心肌缺血再灌注后的无复流现象。
Objective To observe the effect of electroacupuncture pretreatment on myocardial no-reflow after ischemia-reperfusion, NF-?B p65 and its phosphorylation levels in rats. Method Forty-eight male SD rats were randomized to sham operation, model, electroacupuncture pretreatment and atorvastatin pretreatment groups. A model of myocardial no-reflow after acute ischemia-reperfusion was made in all the groups except the sham operation group.The model was made by left coronary artery ligation for 45 min and subsequent reperfusion for 120 min. The extents of myocardial infarct, ischemia and no-reflow were assessed in every group of rats. Serum inflammatory factors interleukin-10(IL-10), interleukin-6(IL-6) and tumor necrosis factor(TNF-?) levels were measured by enzyme-linked immunosorbent assay(ELISA). Myocardial expressions of NF-?B p65 and phospho-NF-?B p65 were determined by Western blot. Result The extents of myocardial infarct and no-reflow were significantly reduced in the electroacupuncture pretreatment and atorvastatin pretr eatment groups compared with the model group.Electroacupuncture pretreatment could significantly raise serum IL-10 levels, reduce serum IL-6 and TNF-? levels and down-regulate myocardial expressions of NF-?B p65 and phospho-NF-?B p65 with statistically significant differences(P<0.01, P<0.05). Conclusion Electroacupuncture pretreatment can significantly decrease myocardial expressions of NF-?B p65 and phospho-NF-?B p65 to reduce myocardial no-reflow after ischemia-reperfusion.
Objective To observe the effect of electroacupuncture pretreatment on myocardial no-reflow after ischemia-reperfusion, NF-?B p65 and its phosphorylation levels in rats. Method Forty-eight male SD rats were randomized to sham operation, model, electroacupuncture pretreatment and atorvastatin pretreatment groups. A model of myocardial no-reflow after acute ischemia-reperfusion was made in all the groups except the sham operation group.The model was made by left coronary artery ligation for 45 min and subsequent reperfusion for 120 min. The extents of myocardial infarct, ischemia and no-reflow were assessed in every group of rats. Serum inflammatory factors interleukin-10(IL-10), interleukin-6(IL-6) and tumor necrosis factor(TNF-?) levels were measured by enzyme-linked immunosorbent assay(ELISA). Myocardial expressions of NF-?B p65 and phospho-NF-?B p65 were determined by Western blot. Result The extents of myocardial infarct and no-reflow were significantly reduced in the electroacupuncture pretreatment and atorvastatin pretr eatment groups compared with the model group.Electroacupuncture pretreatment could significantly raise serum IL-10 levels, reduce serum IL-6 and TNF-? levels and down-regulate myocardial expressions of NF-?B p65 and phospho-NF-?B p65 with statistically significant differences(P<0.01, P<0.05). Conclusion Electroacupuncture pretreatment can significantly decrease myocardial expressions of NF-?B p65 and phospho-NF-?B p65 to reduce myocardial no-reflow after ischemia-reperfusion.
引文
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[12]周全,黄锋.心肌缺血再灌注后无复流发生的炎症机制[J].海南医学,2013,24(17):2559-2562.
[13]王喜丰,汪敏,李刚,等.中性鞘磷脂酶-2(Neutral sphingomyelin-2 N-SMase2)对大鼠脑缺血再灌注损伤中IL-6、IL-1?、TNF-?炎性因子的影响[J].中风与神经疾病杂志,2018,35(4):310-313.
[14]李平,金哈布日,张青山,等.蒙药黄连-4汤对心肌缺血再灌注损伤SD大鼠炎症因子TNF-?、IL-6的影响[J].中西医结合心血管病电子杂志,2017,5(33):124-125.
[15]申冬冬,袁飞,侯江红.黄芪多糖对幼鼠肠缺血再灌注损伤肠组织TNF-?、ICAM-1、IL-6及免疫功能的影响[J].中华中医药学刊,2017,35(6):1528-1532.
[16]王保华.白藜芦醇对大鼠脊髓缺血再灌注损伤组织中TNF-?和IL-6水平的影响[J].中国实用神经疾病杂志,2016,19(12):100-101.
[17]李卉,马敏,咸淑悦,等.加兰他敏对心肌缺血再灌注损伤大鼠血清CK-MB活性及TNF-?和IL-6水平的影响[J].贵阳医学院学报,2016,41(3):280-283.
[18]黄烨,王宗仁,解娟,等.芪丹通脉片对心肌缺血/再灌注大鼠白细胞介素-10的影响[J].中国中医药信息杂志,2008,15(4):38-40.
[19]张洋,王明春,阿日棍,等.丹参(冻干)粉针对大鼠心肌缺血/再灌注损伤的IL-6和IL-10的影响[J].中医药学报,2016,44(3):30-32.
[20]Cheng YS,Dai DZ,Ji H,et al.Sildenafil and FDP-Sr attenuate diabetic cardiomyopathy by suppressing abnormal expression of myocardial CASQ2,FKBP12.6,and SERCA2a in rats[J].Acta Pharmacol Sin,2011,32(4):441-448.
[21]吕翠,王翠花,曾现伟.IL-10基因修饰的BMSCs对大鼠脑缺血再灌注损伤的保护作用[J].实用医学杂志,2016,32(21):3520-3523.
[22]赵红,聂志伟,朱丽娜,等.高压氧对缺血再灌注小鼠脑组织细胞因子IL-10及血脑屏障通透性的影响[J].中国康复医学杂志,2008,23(2):110-113.
[23]陈少宗.针灸双向良性调节作用规律的3种形式[J].针灸临床杂志,2010,26(12):62-65.
[24]李颖,石磊,张宁,等.针刺双向调节效应研究的文献计量分析[J].辽宁中医杂志,2017,44(11):2394-2397,2464.
[25]王旭,王萌萌,李镜,等.针灸“双向调节”作用机制探讨[J].四川中医,2017,35(10):40-42.
[26]张丽丽,刘荔娟,李瑛,等.略论针灸双向调节作用[J].辽宁中医杂志,2013,40(5):997-998.
[27]宋春燕,赵宇辉,马丹,等.针刺预处理对心肌缺血再灌注损伤大鼠血清中CK含量的影响[J].针灸临床杂志,2006,22(10):46-47.
[28]熊英,顾一煌,吴云川.针灸预处理的心肌保护研究进展及应用前景[J].中国运动医学杂志,2012,31(10):926-930,918.
[29]董振华,方剑乔,邵晓梅.针灸防治缺血再灌注损伤机制的研究进展[J].上海针灸杂志,2011,30(12):877-880.
[30]李万山,钟敏,杨进辉,等.电针“内关”穴预处理对缺血再灌注大鼠心肌阿片受体基因表达的影响[J].世界针灸杂志,2012,22(1):43-48.
[31]宋春华,王爽,杨龙.电针心俞穴预处理对心肌缺血再灌注实验大鼠血清NO、CAT的影响[J].上海针灸杂志,2012,31(4):277-279.
[32]王贺,周亚滨.穴位贴敷疗法治疗冠心病不稳定性心绞痛气虚血瘀型临床观察[J].辽宁中医药大学学报,2017,19(4):109-111.
[33]Huang F,Huang JP,Pan JY,et al.Intracellular heat shock protein 70:a possible therapeutic target for preventing postoperative atrial fibrillation[J].Pharmazie,2012,67(9):747-755.
[34]Ling H,Gray CB,Zambon AC,et al.Ca2﹢/Calmodulindependent protein kinaseⅡ?mediates myocardial ischemia/reperfusion injury through nuclear factor-?B[J].Circ Res,2013,112(6):935-944.
[35]丁丹,焦丽华,王雪臣,等.灯盏花素对大鼠心肌缺血再灌注损伤心肌细胞凋亡及NF-κB通路信号分子α7nAChR、p65、IkB-α的影响[J].中国循证心血管医学杂志,2018,10(12):1480-1483,1487.
[36]师文娟,赵咏梅,戚智锋,等.常压高浓度氧对脑缺血-再灌注大鼠缺血核心区核转录因子-κB的适度调节作用[J].首都医科大学学报,2018,39(3):349-354.
[37]朱怡,李毅,张楠,等.电针预处理对心肌缺血再灌注损伤大鼠“内关”穴区Toll样受体4、髓样分化因子88及核转录因子-κB基因表达的影响[J].针刺研究,2018,43(5):302-306,329.
[2]Harrison RW,Aggarwal A,Ou FS,et al.Incidence and outcomes of no-reflow phenomenon during percutaneous coronary intervention among patients with acute myocardial infarction[J].Am J Cardiol,2013,111(2):178-184.
[3]王超,张佳丽,刘昭,等.电针内关穴预处理对兔心肌缺血再灌注CK及心肌细胞活性的影响[J].上海针灸杂志,2015,34(1):67-69.
[4]谭成富,严洁,王超,等.针灸预处理对心肌缺血再灌注损伤兔不同时间热休克蛋白27、70、90表达的影响[J].针刺研究,2017,42(1):31-38.
[5]林海波,严洁,常小荣,等.针灸内关穴预处理对兔缺血再灌注损伤心肌细胞死亡受体通路Fas/FasL蛋白表达的影响[J].中华中医药杂志,2013,28(5):1286-1290.
[6]史晓萍,宗阿南,陶钧,等.《关于善待实验动物的指导性意见》的研究[J].中国医科大学学报,2007,36(4):493.
[7]林文注,王佩.实验针灸学[M].上海:上海科学技术出版社,1999:288.
[8]韦星,张冯迪,阳玉中,等.大鼠心肌缺血再灌注损伤后无复流模型的制备[J].中国动脉硬化杂志,2017,25(7):728-731.
[9]高俊甲.阿托伐他汀对大鼠急性心肌缺血-再灌注无复流现象的影响[J].临床急诊杂志,2018,19(1):30-32.
[10]张晓兰.曲美他嗪联合阿托伐他汀治疗冠心病疗效分析[J].江苏医药,2015,41(14):1723-1724.
[11]陈艺,颜红兵.心肌缺血再灌注后无复流发生的分子机制[J].中国介入心脏病学杂志,2011,19(3):168-171.
[12]周全,黄锋.心肌缺血再灌注后无复流发生的炎症机制[J].海南医学,2013,24(17):2559-2562.
[13]王喜丰,汪敏,李刚,等.中性鞘磷脂酶-2(Neutral sphingomyelin-2 N-SMase2)对大鼠脑缺血再灌注损伤中IL-6、IL-1?、TNF-?炎性因子的影响[J].中风与神经疾病杂志,2018,35(4):310-313.
[14]李平,金哈布日,张青山,等.蒙药黄连-4汤对心肌缺血再灌注损伤SD大鼠炎症因子TNF-?、IL-6的影响[J].中西医结合心血管病电子杂志,2017,5(33):124-125.
[15]申冬冬,袁飞,侯江红.黄芪多糖对幼鼠肠缺血再灌注损伤肠组织TNF-?、ICAM-1、IL-6及免疫功能的影响[J].中华中医药学刊,2017,35(6):1528-1532.
[16]王保华.白藜芦醇对大鼠脊髓缺血再灌注损伤组织中TNF-?和IL-6水平的影响[J].中国实用神经疾病杂志,2016,19(12):100-101.
[17]李卉,马敏,咸淑悦,等.加兰他敏对心肌缺血再灌注损伤大鼠血清CK-MB活性及TNF-?和IL-6水平的影响[J].贵阳医学院学报,2016,41(3):280-283.
[18]黄烨,王宗仁,解娟,等.芪丹通脉片对心肌缺血/再灌注大鼠白细胞介素-10的影响[J].中国中医药信息杂志,2008,15(4):38-40.
[19]张洋,王明春,阿日棍,等.丹参(冻干)粉针对大鼠心肌缺血/再灌注损伤的IL-6和IL-10的影响[J].中医药学报,2016,44(3):30-32.
[20]Cheng YS,Dai DZ,Ji H,et al.Sildenafil and FDP-Sr attenuate diabetic cardiomyopathy by suppressing abnormal expression of myocardial CASQ2,FKBP12.6,and SERCA2a in rats[J].Acta Pharmacol Sin,2011,32(4):441-448.
[21]吕翠,王翠花,曾现伟.IL-10基因修饰的BMSCs对大鼠脑缺血再灌注损伤的保护作用[J].实用医学杂志,2016,32(21):3520-3523.
[22]赵红,聂志伟,朱丽娜,等.高压氧对缺血再灌注小鼠脑组织细胞因子IL-10及血脑屏障通透性的影响[J].中国康复医学杂志,2008,23(2):110-113.
[23]陈少宗.针灸双向良性调节作用规律的3种形式[J].针灸临床杂志,2010,26(12):62-65.
[24]李颖,石磊,张宁,等.针刺双向调节效应研究的文献计量分析[J].辽宁中医杂志,2017,44(11):2394-2397,2464.
[25]王旭,王萌萌,李镜,等.针灸“双向调节”作用机制探讨[J].四川中医,2017,35(10):40-42.
[26]张丽丽,刘荔娟,李瑛,等.略论针灸双向调节作用[J].辽宁中医杂志,2013,40(5):997-998.
[27]宋春燕,赵宇辉,马丹,等.针刺预处理对心肌缺血再灌注损伤大鼠血清中CK含量的影响[J].针灸临床杂志,2006,22(10):46-47.
[28]熊英,顾一煌,吴云川.针灸预处理的心肌保护研究进展及应用前景[J].中国运动医学杂志,2012,31(10):926-930,918.
[29]董振华,方剑乔,邵晓梅.针灸防治缺血再灌注损伤机制的研究进展[J].上海针灸杂志,2011,30(12):877-880.
[30]李万山,钟敏,杨进辉,等.电针“内关”穴预处理对缺血再灌注大鼠心肌阿片受体基因表达的影响[J].世界针灸杂志,2012,22(1):43-48.
[31]宋春华,王爽,杨龙.电针心俞穴预处理对心肌缺血再灌注实验大鼠血清NO、CAT的影响[J].上海针灸杂志,2012,31(4):277-279.
[32]王贺,周亚滨.穴位贴敷疗法治疗冠心病不稳定性心绞痛气虚血瘀型临床观察[J].辽宁中医药大学学报,2017,19(4):109-111.
[33]Huang F,Huang JP,Pan JY,et al.Intracellular heat shock protein 70:a possible therapeutic target for preventing postoperative atrial fibrillation[J].Pharmazie,2012,67(9):747-755.
[34]Ling H,Gray CB,Zambon AC,et al.Ca2﹢/Calmodulindependent protein kinaseⅡ?mediates myocardial ischemia/reperfusion injury through nuclear factor-?B[J].Circ Res,2013,112(6):935-944.
[35]丁丹,焦丽华,王雪臣,等.灯盏花素对大鼠心肌缺血再灌注损伤心肌细胞凋亡及NF-κB通路信号分子α7nAChR、p65、IkB-α的影响[J].中国循证心血管医学杂志,2018,10(12):1480-1483,1487.
[36]师文娟,赵咏梅,戚智锋,等.常压高浓度氧对脑缺血-再灌注大鼠缺血核心区核转录因子-κB的适度调节作用[J].首都医科大学学报,2018,39(3):349-354.
[37]朱怡,李毅,张楠,等.电针预处理对心肌缺血再灌注损伤大鼠“内关”穴区Toll样受体4、髓样分化因子88及核转录因子-κB基因表达的影响[J].针刺研究,2018,43(5):302-306,329.