温针灸对后循环缺血眩晕大鼠神经递质和IKB/NF-κB信号通路的影响

Effects of Warm Acupuncture on Neurotransmitters and IKB/NF-κB Signaling Pathway in Rats with Posterior Circulation Ischemic Vertigo

目的:研究温针灸对后循环缺血眩晕大鼠神经递质和人核因子κB抑制蛋白(inhibitor of NF-κB,IKB)/核转录因子κB(nuclear factor-κB,NF-κB)信号通路的影响。方法:应用随机数字表法将70只SD大鼠随机分为温针灸组、假手术组、模型组、普通针刺组和药物组各14只。假手术组仅分离大鼠右侧颈总动脉(common carotid artery,CCA)及右侧锁骨下动脉(subclavian artery,SCA),不进行CCA及SCA结扎术,直接缝合,其余各组进行CCA及SCA结扎建立后循环缺血眩晕大鼠模型。假手术组及模型组不进行干预;药物组给予盐酸氯桂利嗪治疗;普通针刺组在药物组基础上于“风池“”率谷“”百会”穴进行普通针刺;温针灸组在药物组基础上于“风池“”率谷“”百会”穴进行温针灸。干预结束后,采用激光多普勒血流仪检测各组大鼠前庭神经核血流量下降率;应用TUNEL法检测各组大鼠前庭神经核细胞凋亡指数(apoptotic index,AI);蛋白免疫印迹(western blot,WB)法检测各组大鼠前庭神经核B淋巴细胞因子相关X蛋白(Bcl-2-associated X protein,Bax)、B淋巴细胞瘤2(B cell lymphama,Bcl-2)和含半胱氨酸的天冬氨酸蛋白水解酶3(cysteinyl aspartate specific proteinase 3,caspase-3)蛋白表达;检测各组大鼠脑组织IKB mRNA、NF-κB mRNA及IKB、NF-κB p65蛋白表达;酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测大鼠神经递质变化情况。结果:与模型组比较,药物组、温针灸组和普通针刺组大鼠治疗后右侧前庭神经血流下降率、前庭神经核AI均降低(P<0.05),温针灸组低于普通针刺组和药物组(P<0.05);与模型组比较,药物组、温针灸组和普通针刺组大鼠前庭神经核Bax和Caspase-3蛋白表达降低,Bcl-2蛋白表达升高(P<0.05),温针灸组Bax和Caspase-3蛋白表达低于药物组和普通针刺组(P<0.05),Bcl-2蛋白表达高于药物组和普通针刺组(P<0.05);与模型组比较,药物组、温针灸组和普通针刺组大鼠海马区与大脑皮质中乙酰胆碱酶、谷氨酸浓度降低,去甲肾上腺素、5-羟色胺、γ-氨基丁酸浓度升高(P<0.05),温针灸组乙酰胆碱酶、谷氨酸浓度低于普通针刺组和药物组(P<0.05),5-羟色胺、γ-氨基丁酸高于药物组和普通针刺组(P<0.05);与模型组比较,药物组、温针灸组和普通针刺组大鼠脑组织中IKB、NF-κBp65蛋白及mRNA表达降低(P<0.05),温针灸组低于普通针刺组和药物组(P<0.05)。结论:温针灸能够改善后循环缺血眩晕大鼠眩晕症状,降低神经细胞凋亡,调节神经递质紊乱,其作用机制可能与调控IKB/NF-κB信号通路有关。

Objective:To study the effects of warm acupuncture on neurotransmitters and IKB/NF-κB signaling pathway in rats with posterior circulation ischemic vertigo.Methods:Seventy SD rats were randomly divided into 14 each in the warm acupuncture group,sham operation group,the model group,general acupuncture group and medicine group by applying the random number table method.In the sham-operated group,only SCA and CCA were isolated and directly sutured without ligation surgery,while the rest of the groups were ligated to establish a rat model of ischemic vertigo in the posterior circulation.Sham operation group and the model group were unhandled,the medicine group was given chlorpromazine hydrochloride,the general acupuncture group was treated with general acupuncture at the points of Fengchi(GB20),Shuaigu(GB8)and Baihui(GV20)on the basis of the therapy the drug group accepted,and the warm acupuncture group accepted warm acupuncture at the points of Fengchi,Shuaigu and Baihui on the basis of the medicine group.To detect blood flow in the vestibular nucleus of rats in each group by the end of intervention;to measure apoptotic index(AI)of the vestibular nucleus of rats in each group using TUNEL method;to test the expressions of Bax,Bcl-2 and cysteine aspartate protease-3(Caspase-3)protein in the vestibular nucleus in each group;to detect the expressions of IKB mRNA,NF-κBmRNA and IKB,NFκBp65 protein expression in the brain tissues of rats in each group;and to observe the changes of neurotransmitter in rats.Results:Compared with the model group,the rate of decrease in blood flow in the right vestibular nerve and AI in the vestibular nerve nucleus were decreased in the rats in the drug group,the warm acupuncture group and the common acupuncture group after treatment(P<0.05),and the warm acupuncture group was lower than the common acupuncture group and the drug group(P<0.05);compared with the model group,the expression of Bax and Caspase-3 proteins in the vestibular nerve nucleus of the rats in the drug group,the warm acupuncture group and the common acupuncture group was decreased and the expression of Bcl-2 protein was elevated(P<0.05),and Bax and Caspase-3 protein expression in the warm acupuncture group was lower than that in the drug group and the common acupuncture group(P<0.05),and Bcl-2 protein expression was higher than that in the drug group and the common acupuncture group(P<0.05);compared with the model group,the concentrations of norepinephrine and 5-hydroxytryptamine decreased while acetylcholinesterase,glutamate,γ-aminobutyric acid increased in the hippocampal region and cerebral cortex of rats in the drug group,the warm acupuncture group and the common acupuncture group(P<0.05);norepinephrine and 5-hydroxytryptamine concentration in the warm acupuncture group was lower than that in the common acupuncture group and the drug group(P<0.05),and acetylcholinesterase,glutamate,andγ-aminobutyric acid were higher than that in the drug group and the common acupuncture group(P<0.05);in comparison to the model group,the expressions of IKB,NF-κBp65 protein and its mRNA reduced in the brain tissue of rats in the drug group,warm acupuncture group and common acupuncture group(P<0.05),and the warm acupuncture group was lower than the common acupuncture and drug groups(P<0.05).Conclusion:Warm acupuncture can improve the symptom of the rat models,reduce neuronal apoptosis,and regulate neurotransmitter disorders in rats with posterior circulation ischemic vertigo,and its mechanism of action may be related to the regulation of IKB/NF-κB signaling pathway.