针刺风池、外关、阳陵泉对偏头痛模型大鼠NF-κB P65、IKKβ、IκBα的影响

Effect of Acupuncture at“Fengchi”(GB20),“Waiguan”(SJ5)and“Yanglingquan”(GB34)on NF-κB P65,IKKβ,IκBαin Migraine Rats

目的观察针刺风池、外关、阳陵泉对硝酸甘油(NTG)诱导的偏头痛(ME)大鼠核因子-κB(NF-κB)P65、IκB激酶(IKK)β、NF-κB抑制蛋白(IκB)α的影响,探讨针刺治疗ME可能的作用机制。方法将40只SD大鼠随机分为空白组、模型组、针刺组、西药组,每组10只。空白组不做任何处理;模型组采用皮下注射NTG造模;针刺组造模后给予针刺风池、外关、阳陵泉干预治疗8 d;西药组造模后给予盐酸氟桂利嗪药液灌胃干预治疗8 d。观察并记录各组大鼠造模前、造模后、治疗后的行为学变化以及50%机械缩足阈值(PWT);采用蛋白免疫印迹(WB)法检测各组大鼠脑膜NF-κB P65、IKKβ、IκBα的蛋白表达;反转录-聚合酶链反应(RT-PCT)法检测各组大鼠三叉神经脊束核NF-κB P65、IKKβ、IκBα的mRNA表达。结果(1)造模后,除空白组外其余3组大鼠均出现耳红、咬尾、爬笼等行为,行为学评分增加,50%PWT降低(P<0.01),表明造模成功。(2)造模后,与空白组比较,模型组大鼠行为学的评分均明显升高(P<0.01);治疗后,与模型组比较,针刺组和西药组行为学评分明显降低(P<0.01);与西药组比较,针刺组差异不明显(P>0.05)。(3)造模后,与空白组比较,其余3组大鼠50%PWT均显著降低(P<0.01);治疗后,与模型组比较,针刺组和西药组50%PWT明显升高(P<0.01);与西药组比较,针刺组差异不明显(P>0.05)。(4)与空白组比较,模型组NF-κB P65、IKKβ、IκBα的蛋白含量明显升高(P<0.01);与模型组比较,针刺组与西药组NF-κB P65、IKKβ、IκBα的蛋白含量明显降低(P<0.01),其中针刺组与西药组差异无统计学意义(P>0.05)。(5)与空白组比较,模型组NF-κB P65、IKKβ、IκBα的mRNA表达明显升高(P<0.01);与模型组比较,针刺组与西药组NF-κB P65、IKKβ、IκBα的mRNA表达明显降低(P<0.01),其中针刺组与西药组差异无统计学意义(P>0.05)。结论针刺风池、外关、阳陵泉能够减轻ME大鼠的疼痛程度,有效下调脑膜及三叉神经脊束核中NF-κB P65、IKKβ、IκBα的蛋白含量与基因表达,这可能是针刺风池、外关、阳陵泉治疗ME的机制之一。

Objective To observe the effects of acupuncture at“Fengchi”(GB20),“Waiguan”(SJ5)and“Yanglingquan”(GB34)on NF-κB p65,IKKβand IκBαin migraine(ME)rats induced by nitroglycerin(NTG),and to explore the possible mechanism of acupuncture in treating ME.Methods 40 SD rats were randomly divided into blank group,model group,acupuncture group and western medicine group,with 10 rats in each group.The blank group was not given any treatment;the model group was modeled by subcutaneous injection of NTG,the acupuncture group was intervened treatment by acupuncture at“Fengchi”(GB20),“Waiguan”(SJ5)and“Yanglingquan”(GB34)for 8 days after modeling,the western medicine group was intervened treatment by intragastric administration of flunarizine hydrochloride for 8 days after modeling.Behavioral changes and 50%mechanical shrinkage threshold(PWT)were observed and recorded before modeling,after modeling and after treatment.The protein expression levels of NF-κB P65,IKKβand IκBαin meninges were detected by Western blot(WB).The mRNA content of NF-κB P65,IKKβand IκBαin spinal trigeminal nucleus was detected by reverse transcription-polymerase chain reaction(RT-PCR).Results(1)After modeling,except for the blank group,the other three groups all had red ears,tail biting and cage climbing behaviors,the behavioral score increased and 50%PWT decreased(P<0.01),indicating the success of modeling.(2)After modeling,the behavioral scores of rats in model group were significantly higher than those in the blank group(P<0.01);after treatment,compared with the model group,the behavioral scores of the acupuncture group and the western medicine group were significantly lower(P<0.01);compared with the western medicine group,there was no significant difference in the acupuncture group(P>0.05).(3)After modeling,except for the blank group,the 50%PWT of rats in the other three groups was significantly lower than that in the blank group(P<0.01);after treatment,compared with the model group,the 50%PWT in the acupuncture group and the western medicine group was significantly increased(P<0.01);compared with the western medicine group,there was no significant difference in the acupuncture group(P>0.05).(4)The content of NF-κB P65,IKKβand IκBαprotein in the model group was significantly higher than those in the blank group(P<0.01);the protein content of NF-κB P65,IKKβand IκBαin acupuncture group and western medicine group were significantly decreased(P<0.01),and there was no significant difference between acupuncture group and western medicine group(P>0.05).(5)Compared with the blank group,the mRNA levels of NF-κB P65,IKKβand IκBαin the model group were significantly higher(P<0.01);compared with model group,the mRNA levels of NF-κB P65,IKKβand IκBαin the acupuncture group and western medicine group were significantly decreased(P<0.01),and there was no significant difference between acupuncture group and western medicine group(P>0.05).Conclusion Acupuncture at“Fengchi”(GB20),“Waiguan”(SJ5)and“Yanglingquan”(GB34)can relieve the pain of ME rats,effectively reduce the protein and gene expression of NF-κB P65,IKKβand IκBαin the meninges and trigeminal spinal tract nucleus,which may be one of the mechanisms of acupuncture at“Fengchi”(GB20),“Waiguan”(SJ5)and“Yanglingquan”(GB34)in the treatment of ME.