“标本配穴”电针预处理抗脑缺血再灌注损伤的效应研究
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摘要
目的从腧穴配伍角度,比较"标本配穴"电针与常规配穴电针预处理对脑缺血再灌注损伤模型大鼠的防治效应,观察"标本配穴"电针预处理抗脑缺血再灌注损伤的可能优势作用,为临床腧穴配伍提供参考依据。方法 SPF级雄性SD大鼠100只,随机分为假手术组、模型组、标本配穴组、常规配穴组,每组各25只。模型组采用改良的MCAO线栓法缺血2h后行再灌注,制备大鼠右侧局灶性脑缺血再灌注损伤模型。两电针组造模前取相应腧穴接HANS-200电针仪预处理,疏密波,频率2/100Hz,强度1mA,每15min为1个刺激单元(通电10min,留针5min),连续重复4次,共计1h。脑缺血再灌后3h对各组进行神经行为学评分,用TTC法检测梗死百分比,检测海马组织丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSHPx)的活性,HE染色观察神经元形态变化。结果电针预处理后两电针组行为学评分、梗死百分比和MDA含量均较模型组降低(P<0.01),SOD活性值和GSH-Px活性值均较模型组升高(P<0.01),海马神经元损伤较模型组减轻;两电针组之间比较,标本配穴组行为学评分、梗死百分比和MDA含量低于常规配穴组(P<0.05),SOD活性值及GSH-Px活性值均高于常规配穴组(P<0.01),海马神经元损伤程度较常规配穴组轻。结论 "标本配穴"电针预处理可能是通过激活大量抗自由基酶,减轻大量自由基对神经元的氧化应激损伤抗脑缺血再灌注损伤,且这种抗损伤效应优于常规配穴电针预处理。
Objective To compare the effect of pretreatment between "combined biao and ben acupoints" and routine Electro-acupuncture(EA) preconditioning on rats of cerebral ischemia-reperfusion injury from the aspect of acupoints compatibility,and provide evidence for acupoints compatibility in clinic. Methods A total of 80 SPF SD male rats were randomly divided into sham group,model group, EA1 group(GV20, SP6, BL23), EA2 group(GV20, GV14,GV26). The model was achieved by the method of improved MCAO with ischemia for 2 h. Before having been established model, both EA1 group and EA2 group received related acupoints stimulation by instrument of HANS-200 type with density wave, 2/100 Hz and 1 mA for 10 min, then rest for 5 min, which of this pattern was recycled 4 times. After perfusion for 3 h, the Neurobehavioral score was performed, and then the infarct volume of the brain by TTC, and SOD, MDA, GSH-Px of hippocampus was tested by spectrometer, the changes of neurons morphology in hippocampus of rats were observed. Results The Neurobehavioral score, infarct volume, MDA in both EA1 group and EA2 group were obviously lower than model group(P<0.01), and the level of SOD and GSH-Px were obviously higher than model group(P<0.01),and the degree of neurons damage in both EA1 group and EA2 group were obviously lower than model group. The Neurobehavioral score, infarct volume,MDA in EA1 group were all obviously lower than EA2 group(P<0.05), and the Neurobehavioral score, SOD and GSH-Px were all obviously higher than EA2 group(P<0.05), and the degree of neurons damage in EA1 group was obviously lower than EA2 group. Conclusion The preconditioning with EA1 resists harm of cerebrall ischemia-reperfusion injury through the pathway of activating SOD and GSH-Px to reduce oxyradical,and this ability could be better than EA2.
Objective To compare the effect of pretreatment between "combined biao and ben acupoints" and routine Electro-acupuncture(EA) preconditioning on rats of cerebral ischemia-reperfusion injury from the aspect of acupoints compatibility,and provide evidence for acupoints compatibility in clinic. Methods A total of 80 SPF SD male rats were randomly divided into sham group,model group, EA1 group(GV20, SP6, BL23), EA2 group(GV20, GV14,GV26). The model was achieved by the method of improved MCAO with ischemia for 2 h. Before having been established model, both EA1 group and EA2 group received related acupoints stimulation by instrument of HANS-200 type with density wave, 2/100 Hz and 1 mA for 10 min, then rest for 5 min, which of this pattern was recycled 4 times. After perfusion for 3 h, the Neurobehavioral score was performed, and then the infarct volume of the brain by TTC, and SOD, MDA, GSH-Px of hippocampus was tested by spectrometer, the changes of neurons morphology in hippocampus of rats were observed. Results The Neurobehavioral score, infarct volume, MDA in both EA1 group and EA2 group were obviously lower than model group(P<0.01), and the level of SOD and GSH-Px were obviously higher than model group(P<0.01),and the degree of neurons damage in both EA1 group and EA2 group were obviously lower than model group. The Neurobehavioral score, infarct volume,MDA in EA1 group were all obviously lower than EA2 group(P<0.05), and the Neurobehavioral score, SOD and GSH-Px were all obviously higher than EA2 group(P<0.05), and the degree of neurons damage in EA1 group was obviously lower than EA2 group. Conclusion The preconditioning with EA1 resists harm of cerebrall ischemia-reperfusion injury through the pathway of activating SOD and GSH-Px to reduce oxyradical,and this ability could be better than EA2.
引文
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[12]李强,徐芬,梁凤霞,等.“标本配穴”法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[J].中国针灸,2014,34(6):578-582.
[13]陈丽,梁凤霞,陈瑞,等.“标本配穴”针灸结合甲钴胺治疗糖尿病周围神经病变的临床对照研究[J].中国中西医结合消化杂志,2014,22(4):204-207.
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[16]宋艳,赵国桢,赵百孝,等.不同时机介入电针对模拟失重大鼠肝脏HSP 70、MDA、SOD和GSH-Px的影响[J].针刺研究,2015,40(5):383-387.
[17]郭健,刘义,李延平,等.氧自由基与心肌缺血再灌注损伤[J].中国心血管杂志,2008,13(5):384-387.
[18]沈梅红,李成,李忠仁.电针对脑缺血再灌注模型大鼠的GSH含量、GSH-Px及GR活性的影响[J].南京中医药大学学报,2011,27(2):137-139.
[19]王富明,张亚敏,孙华,等.针刺对大鼠脑缺血再灌注损伤后不同时间点血清SOD和MDA表达的影响[J].针灸临床杂志,2015,31(2):62-69.
[20]韩丽,赵百孝,刘铜华,等.不同浓度艾烟干预对大鼠肺脏及血清SOD、MDA表达的影响[J].中国针灸,2015,30(7):687-690.
[21]林浴坤,林如辉,陈斌,等.电针对局灶性脑缺血再灌注模型大鼠学习记忆作用的氧化应激机制[J].中国康复医学杂志,2015,30(8):755-760.
[22]张继苹,曲姗姗,吴春晓,等.穴位配伍的脑功能成像研究现状[J].针灸临床杂志,2013,29(1):73-76.
[23]张晓琳,刘智艳,刘娟,等.电针对老年性痴呆大鼠记忆功能减退的影响[J].中华中医药杂志,2012,27(3):706-709.
[24]宋艳,嵇波,汪德生,等.不同时机针刺对模拟失重大鼠肾功能和肾脏氧自由基代谢的影响[J].中国针灸,2014,34(11):1106-1110.
[2]王占奎,倪光夏,刘坤,等.脑缺血再灌注大鼠白细胞介素-1受体与肿瘤坏死因子受体变化及针刺干预的时效性研究[J].中国针灸,2012,32(11):1012-1018.
[3]谢亚宁,王枫,王强,等.电针对脑缺血再灌注大鼠脑红蛋白的影响[J].针刺研究,2012,37(5):380-384.
[4]谢俊,陈泽斌,吴松,等.不同腧穴配伍电针防治大鼠心肌缺血损伤作用的比较[J].针刺研究,2017,42(2):131-135.
[5]王华,梁凤霞.腧穴配伍研究思路和展望[J].中国针灸,2012,32(4):359-362.
[6] Longa EZ,Weinstein PR,Carlson S,et al. Reversible middlecerebral arteryocclusion without craniectomy in rats[J]. Stro-ke,1989,20(l):84-91.
[7]华兴邦,李辞蓉,周浩良,等.大鼠穴位图谱的研制[J].实验动物与动物实验,1991,(1):1-5.
[8] Kato H,Kogure K. Biochemical and molecular characteris-tics of the brain withdeveloping cerebral infarction[J]. CellMol Neurobilo,1999,19(1):93-108.
[9]陈怀龙,齐慧,刘孝洁,等.电针预处理对全脑缺血/再灌注损伤大鼠海马葡萄糖调节蛋白78表达的影响[J].中国针灸,2014,34(9):889-893.
[10]林咸明,陈丽萍,姚旭.不同时程电针预处理对脑缺血再灌注大鼠血脑屏障基质金属蛋白酶-9、血管内皮生长因子的影响[J].针刺研究,2015,40(1):40-44.
[11]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[J].中医杂志,2014,55(4):328-332.
[12]李强,徐芬,梁凤霞,等.“标本配穴”法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[J].中国针灸,2014,34(6):578-582.
[13]陈丽,梁凤霞,陈瑞,等.“标本配穴”针灸结合甲钴胺治疗糖尿病周围神经病变的临床对照研究[J].中国中西医结合消化杂志,2014,22(4):204-207.
[14]郝青,吴松,刘建民,等.“标本配穴”法电针对老年阳虚模型大鼠垂体-靶腺轴的影响[J].中国针灸,2014,34(10):993-997.
[15] Kikuchi K,Kawahara K,Tanchareon S,et al. The free rad-ical scavenger rescuesrats from cerebral infarction by at-tenuating the release of high-mobility groupbox-l in neur-onal cells[J]. J pharmacol Exp Ther,2009,329(3):865-874.
[16]宋艳,赵国桢,赵百孝,等.不同时机介入电针对模拟失重大鼠肝脏HSP 70、MDA、SOD和GSH-Px的影响[J].针刺研究,2015,40(5):383-387.
[17]郭健,刘义,李延平,等.氧自由基与心肌缺血再灌注损伤[J].中国心血管杂志,2008,13(5):384-387.
[18]沈梅红,李成,李忠仁.电针对脑缺血再灌注模型大鼠的GSH含量、GSH-Px及GR活性的影响[J].南京中医药大学学报,2011,27(2):137-139.
[19]王富明,张亚敏,孙华,等.针刺对大鼠脑缺血再灌注损伤后不同时间点血清SOD和MDA表达的影响[J].针灸临床杂志,2015,31(2):62-69.
[20]韩丽,赵百孝,刘铜华,等.不同浓度艾烟干预对大鼠肺脏及血清SOD、MDA表达的影响[J].中国针灸,2015,30(7):687-690.
[21]林浴坤,林如辉,陈斌,等.电针对局灶性脑缺血再灌注模型大鼠学习记忆作用的氧化应激机制[J].中国康复医学杂志,2015,30(8):755-760.
[22]张继苹,曲姗姗,吴春晓,等.穴位配伍的脑功能成像研究现状[J].针灸临床杂志,2013,29(1):73-76.
[23]张晓琳,刘智艳,刘娟,等.电针对老年性痴呆大鼠记忆功能减退的影响[J].中华中医药杂志,2012,27(3):706-709.
[24]宋艳,嵇波,汪德生,等.不同时机针刺对模拟失重大鼠肾功能和肾脏氧自由基代谢的影响[J].中国针灸,2014,34(11):1106-1110.