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

Effects of Acupuncture at Baihui(GV 20) and Sishencong(EX-HN 1) on the Effects of the NF-κB signaling pathway in CI/RI model rats

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.