针刺百会、四神聪对脑缺血再灌注损伤模型大鼠缺血半暗带组织NF-κB信号通路的影响
|
摘要
目的采用针刺百会、四神聪穴位的方法,干预脑缺血再灌注损伤模型大鼠,通过对各组大鼠脑缺血半暗带组织中NF-κB、IκB蛋白表达水平的检测,探讨针刺百会、四神聪治疗缺血性脑病的部分作用机制。方法SPF级SD大鼠45只,将大鼠随机分为2组:假手术组(15只);造模组(30只)。然后采用改良线栓法对造模组30只大鼠进行CIRI模型复制,评价模型成功后,再将成功的动物模型随机分为2组:非穴区对照组和针刺组。最终实验分为3组,即:假手术组、非穴区对照组和针刺组。假手术组大鼠采用相同术式但不予线栓干预。模型复制成功后,针刺组大鼠给予百会、四神聪穴针刺干预,间隔8 h 1次,至再灌注72 h结束;非穴区对照组大鼠分别于百会、四神聪穴区周边5 mm处进行针刺皮肤干预,间隔8 h 1次,至再灌注72 h结束。末次针刺后1 h,过量麻醉法处死大鼠,取脑组织,去除缺血液化区组织后,沿缺血灶走行,剥离缺血灶周边2 mm区域内半暗带组织,用于NF-κB、IκB蛋白表达水平的检测。结果假手术组神经功能缺损评分为0,TTC染色未见梗死灶。非穴区对照组和针刺组神经功能缺损评分和梗死百分比均具有显著的统计学差异。与假手术组比较,非穴区对照组和针刺组大鼠缺血半暗带组织中NF-κB表达水平显著上调(P<0.01),IκB蛋白表达水平显著下降(P<0.01),有统计学差异;针刺干预后,与非穴区对照组比较,针刺组大鼠缺血半暗带组织中NF-κB表达水平显著下调(P<0.01),IκB蛋白表达水平显著上调(P<0.01),有统计学差异。结论针刺百会、四神聪穴可明显降低脑缺血再灌注损伤模型大鼠神经功能缺损评分,减小梗死百分比。其作用机制可能是通过调节NF-κB信号通路实现的。
Objective The method of acupuncture at Baihui(GV 20) and Sishencong(EX-HN 1) was used to interfere with the cerebral ischemia-reperfusion injury model rats.The expression level of NF-κB and IκB protein in the ischemic penumbra of rats in each group was detected.To explore the mechanism of acupuncture Baihui(GV 20) and Sishencong(EX-HN 1) in the treatment of ischemic encephalopathy.Methods 45 of the SPF SD rats were randomly divided into sham operation group(n=15) and molding group(n=30).Then,the CIRI model was used to replicate the CIRI model of 30 rats in the molding group.After the success of the evaluation model,the successful animal models were randomly divided into non-acupoint control group and the acupuncture group.The final experiment was divided into the sham operation group,the non-acupoint control group and the acupuncture group.The rats in the sham group underwent the same operation but did not intervene.After the success of the model replication,the rats in the acupuncture group were given Baihui(GV 20) and Sishencong(EX-HN 1) acupuncture points,and the interval was 8 hours,and the time was completed by 72 hours.The rats in the control non-acupoint group were treated with acupuncture for 5 mm around Baihui(GV 20) and Sishencong(EX-HN 1),once every 8 hours,until the end of 72 hours of reperfusion.One hour after the last acupuncture,the rats were killed by excessive anesthesia,the brain tissue was removedwalked along the ischemic zone,and the penumbra tissue around the ischemic lesion was stripped out for the detection of the expression of NF-κB and IκB protein.Results The nerve function defect of the sham operation group was 0,no infarct by TTC staining.The scores of nerve function defects and the percentage of infarction were statistically significant in the control group and the acupuncture group.Compared with the sham operation group,the expression levels of NF-κB protein were significantly increased in non-acupoint control group and acupuncture group(P<0.01),and IκB protein were significantly reduced(P<0.01) with statistical difference.Compared with the control group,the expression level of NF-κB protein in the ischemic semi-dark bands of acupuncture group was significantly reduced(P<0.01),and IκB was increased(P<0.01),with statistical difference.Conclusion The acupuncture Baihui(GV 20) and Sishencong(EX-HN 1) acupoints can significantly reduce the neural function defect score of cerebral ischemia reperfusion model and decrease the percentage of infarction.The mechanism may be realized by regulating the NF-κB signaling pathway.
Objective The method of acupuncture at Baihui(GV 20) and Sishencong(EX-HN 1) was used to interfere with the cerebral ischemia-reperfusion injury model rats.The expression level of NF-κB and IκB protein in the ischemic penumbra of rats in each group was detected.To explore the mechanism of acupuncture Baihui(GV 20) and Sishencong(EX-HN 1) in the treatment of ischemic encephalopathy.Methods 45 of the SPF SD rats were randomly divided into sham operation group(n=15) and molding group(n=30).Then,the CIRI model was used to replicate the CIRI model of 30 rats in the molding group.After the success of the evaluation model,the successful animal models were randomly divided into non-acupoint control group and the acupuncture group.The final experiment was divided into the sham operation group,the non-acupoint control group and the acupuncture group.The rats in the sham group underwent the same operation but did not intervene.After the success of the model replication,the rats in the acupuncture group were given Baihui(GV 20) and Sishencong(EX-HN 1) acupuncture points,and the interval was 8 hours,and the time was completed by 72 hours.The rats in the control non-acupoint group were treated with acupuncture for 5 mm around Baihui(GV 20) and Sishencong(EX-HN 1),once every 8 hours,until the end of 72 hours of reperfusion.One hour after the last acupuncture,the rats were killed by excessive anesthesia,the brain tissue was removedwalked along the ischemic zone,and the penumbra tissue around the ischemic lesion was stripped out for the detection of the expression of NF-κB and IκB protein.Results The nerve function defect of the sham operation group was 0,no infarct by TTC staining.The scores of nerve function defects and the percentage of infarction were statistically significant in the control group and the acupuncture group.Compared with the sham operation group,the expression levels of NF-κB protein were significantly increased in non-acupoint control group and acupuncture group(P<0.01),and IκB protein were significantly reduced(P<0.01) with statistical difference.Compared with the control group,the expression level of NF-κB protein in the ischemic semi-dark bands of acupuncture group was significantly reduced(P<0.01),and IκB was increased(P<0.01),with statistical difference.Conclusion The acupuncture Baihui(GV 20) and Sishencong(EX-HN 1) acupoints can significantly reduce the neural function defect score of cerebral ischemia reperfusion model and decrease the percentage of infarction.The mechanism may be realized by regulating the NF-κB signaling pathway.
引文
[1]马贤德,侯殿东,张威.眼针对MCAO/R大鼠大脑缺血区半暗带脑组织中BDNF和TrkB表达的影响及其脑保护作用机制[J].吉林大学学报(医学版),2014,40(3):549-553.
[2]马贤德,王守岩,张威.眼针对脑缺血再灌注模型大鼠脑组织BDNF、p75NTR表达影响及其脑保护作用机制[J].北京中医药大学学报,2014,37(1):48-52.
[3]王玉,杨洁红,何昱,等.养阴通脑颗粒对大鼠脑缺血/再灌注损伤的保护作用及其抑制NF-κB信号通路的作用机制[J].中华中医药杂志,2017,32(8):3713-3717.
[4]张鑫,耿诗涵,王萍,等.苦碟子注射液对大鼠脑缺血再灌注损伤炎症及TLR-4/NF-κB信号的影响[J].药物评价研究,2017,40(7):892-896.
[5]陈文明,钱民章.绞股蓝总苷对大鼠脑缺血再灌注损伤及NF-κB p65信号通路的影响[J].遵义医学院学报,2017,40(3):273-277.
[6]张莹,秦文熠,罗勇.电针对局灶脑缺血/再灌注大鼠大脑缺血皮质区白介素-4/信号传导及转录激活因子6与核因子-κB的影响[J].针刺研究,2017,42(3):217-222+258.
[7]葛建彬,卢红建,宋新建,等.枸杞多糖对小鼠脑缺血再灌注损伤的保护作用及其抑制NF-κB,TNF-α,IL-6和IL-1β表达的机制[J].中国中药杂志,2017,42(2):326-331.
[8]宋书昌,卢智,陈华,等.梅花针叩刺百会、四神聪治疗急性期脑梗死合并认知功能障碍的临床研究[J].针灸临床杂志,2014,30(1):26-28.
[9]黄伟,李佳,朱广为.针刺百会、人中穴对急性脑缺血大鼠模型NF-κB/IκB-α的影响[J].中华中医药杂志,2017,32(1):298-302.
[10]马贤德,孙宏伟,柴纪严,等.线栓法制备大鼠脑缺血再灌注模型的方法研究[J].中华中医药学刊,2009,27(6):1200-1201.
[11]BEDERSON J B,PITTSL H,TSUJI M,et al.Rat middle cerebral artery occlusion evaluation of the model and development of a neurologic evaluation[J].Stroke,1986,17(3):472-476.
[12]华兴邦.大白鼠穴位图谱的研制[J].实验动物与动物实验,1991(1):1-6.
[13]于船.中国兽医针灸学,北京:农业出版社,1984.
[14]徐叔云,卞如濂.药理实验方法学[M].第3版.北京:人民卫生出版社,2002.
[2]马贤德,王守岩,张威.眼针对脑缺血再灌注模型大鼠脑组织BDNF、p75NTR表达影响及其脑保护作用机制[J].北京中医药大学学报,2014,37(1):48-52.
[3]王玉,杨洁红,何昱,等.养阴通脑颗粒对大鼠脑缺血/再灌注损伤的保护作用及其抑制NF-κB信号通路的作用机制[J].中华中医药杂志,2017,32(8):3713-3717.
[4]张鑫,耿诗涵,王萍,等.苦碟子注射液对大鼠脑缺血再灌注损伤炎症及TLR-4/NF-κB信号的影响[J].药物评价研究,2017,40(7):892-896.
[5]陈文明,钱民章.绞股蓝总苷对大鼠脑缺血再灌注损伤及NF-κB p65信号通路的影响[J].遵义医学院学报,2017,40(3):273-277.
[6]张莹,秦文熠,罗勇.电针对局灶脑缺血/再灌注大鼠大脑缺血皮质区白介素-4/信号传导及转录激活因子6与核因子-κB的影响[J].针刺研究,2017,42(3):217-222+258.
[7]葛建彬,卢红建,宋新建,等.枸杞多糖对小鼠脑缺血再灌注损伤的保护作用及其抑制NF-κB,TNF-α,IL-6和IL-1β表达的机制[J].中国中药杂志,2017,42(2):326-331.
[8]宋书昌,卢智,陈华,等.梅花针叩刺百会、四神聪治疗急性期脑梗死合并认知功能障碍的临床研究[J].针灸临床杂志,2014,30(1):26-28.
[9]黄伟,李佳,朱广为.针刺百会、人中穴对急性脑缺血大鼠模型NF-κB/IκB-α的影响[J].中华中医药杂志,2017,32(1):298-302.
[10]马贤德,孙宏伟,柴纪严,等.线栓法制备大鼠脑缺血再灌注模型的方法研究[J].中华中医药学刊,2009,27(6):1200-1201.
[11]BEDERSON J B,PITTSL H,TSUJI M,et al.Rat middle cerebral artery occlusion evaluation of the model and development of a neurologic evaluation[J].Stroke,1986,17(3):472-476.
[12]华兴邦.大白鼠穴位图谱的研制[J].实验动物与动物实验,1991(1):1-6.
[13]于船.中国兽医针灸学,北京:农业出版社,1984.
[14]徐叔云,卞如濂.药理实验方法学[M].第3版.北京:人民卫生出版社,2002.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |