针刺对脑心综合征大鼠脑组织细胞凋亡、炎症因子及丝裂原活化蛋白激酶/核因子-κB/激活蛋白-1信号通路的影响

The effects of acupuncture on apoptosis of brain tissue cells,inflammatory factors,and mitogen activated protein kinase/nuclear factor-κB/activator protein-1 signaling pathway in rats with cerebral heart syndrome

目的探讨针刺对脑心综合征大鼠脑组织细胞凋亡、炎症因子及丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)/核因子-κB(nuclear factor-κB,NF-κB)/激活蛋白-1(activator protein-1,AP-1)信号通路的影响。方法将36只SD大鼠随机分为针刺组、模型组和对照组,各12只。采用胶原酶加肝素联合注射大鼠尾状核制备脑心综合征大鼠模型,对照组给予等剂量生理盐水向大鼠尾状核注入,手术操作过程同模型组。模型组和假手术组均不予针刺;针刺组选心俞穴、内关穴、风府穴和水沟穴,每日针刺1次,连续3天。采用原位末端凋亡(TdT-mediated dUTP-biotin nick end labeling,TUNEL)法测定脑组织细胞凋亡情况;神经功能采用Zea-Longa法评估;运用酶联免疫法测定白细胞介素(interleukin,IL)-1β、IL-6和肿瘤坏死因子(tumor necrosis factor,TNF)-α水平;采用Western Blot法测定MAPK、NF-κB、AP-1蛋白表达。结果模型组和针刺组大鼠脑组织细胞凋亡指数(apoptosis index,AI)高于假手术组(P<0.05);针刺组大鼠脑组织AI低于模型组(P<0.05)。模型组大鼠神经行为评分高于假手术组(P<0.05);针刺组大鼠神经行为评分低于模型组(P<0.05)。模型组大鼠IL-1β、IL-6和TNF-α水平高于假手术组(P<0.05);针刺组大鼠IL-1β、IL-6和TNF-α水平低于模型组(P<0.05)。模型组大鼠MAPK蛋白灰度值、NF-κB蛋白灰度值和AP-1蛋白灰度值高于假手术组(P<0.05);针刺组大鼠MAPK蛋白灰度值、NF-κB蛋白灰度值和AP-1蛋白灰度值低于模型组(P<0.05)。结论脑心综合征大鼠采用针刺可减轻大鼠脑组织细胞凋亡,减轻大鼠炎症因子,且可下调MAPK/NF-κB/AP-1信号通路表达。

Objective To explore the effects of acupuncture on brain tissue cell apoptosis,inflammatory factors,and mitogen activated protein kinase(MAPK)/nuclear factor-κB(NF-κB)/activator protein-1(AP-1)signaling pathways in rats with cerebral heart syndrome.Methods 36 SD rats were divided into the acupuncture group,the model group and the control group,with 12 rats in each group.A rat model of cerebral heart syndrome was prepared by combining collagenase and heparin caudate nucleus.The control group was given an equal dose of physiological saline injection into the caudate nucleus of rats,and the surgical procedure was the same as that of the model group.Both the model group and the sham surgery group did not receive acupuncture,only once a day during air grabbing;The acupuncture group selected Xinshu point,Neiguan point,Fengfu point,and Shuigou point,and acupuncture was performed once a day for three consecutive days.Measure the apoptosis of brain tissue cells using TdT-mediated dUTP-biotin nick end labeling(TUNEL)method;Neurological function was evaluated using the Zea Longa method;Using enzyme linked immunosorbent assay method to measure the levels of interleukin(IL)-1β,IL-6,and tumor necrosis factor-α(TNF)-α;Using Western Blot method to determine MAPK/NF-κB/AP-1 protein expression.Results The model group and the acupuncture group AI of brain tissue was higher than the sham surgery group(P<0.05);acupuncture group AI of brain tissue was lower than the model group(P<0.05).The model group and the sham surgery group neurobehavioral scores of rats were higher sham surgery group(P<0.05);acupuncture group neurobehavioral scores of rats was lower than the model group(P<0.05).The model group and sham surgery group levels of IL-1β,IL-6,and TNF-αwere higher than the sham surgery group(P<0.05);acupuncture group rats levels of IL-1β,IL-6,and TNF-αwere lower than the model group(P<0.05).The model group gray values of MAPK protein,NF-κB protein,and AP-1 protein were higher than the sham surgery group(P<0.05);the acupuncture group rats gray values of MAPK protein,NF-κB protein,and AP-1 protein were lower than the model group(P<0.05).Conclusion Acupuncture in rats with cerebral heart syndrome can reduce apoptosis of brain tissue cells,alleviate inflammatory factors,and downregulate the expression of MAPK/NF-κB/AP-1 signaling pathway.