摘要
目的:观察电头针对缺血性脑卒中大鼠缺血大脑皮层区过氧化物酶体增殖物激活受体γ(PPARγ)、磷酸化核因子-κB p65(p-NF-κB p65)、促炎及抑炎因子白细胞介素(IL)-1β、IL-4等及小胶质细胞标志物的影响,探讨电头针减轻缺血性脑卒中炎性损伤的机制。方法:SD大鼠随机选取12只作为假手术组,其余采用线栓法复制大脑中动脉栓塞(MCAO)大鼠模型。造模成功的大鼠随机分为模型组、电头针组、电头针+抑制剂组,每组12只。电头针组电针双侧“顶颞前斜线”,每次30 min;电头针+抑制剂组腹腔注射PPARγ抑制剂GW9662溶液(0.1 mg/mL)后,予以电头针干预。各组大鼠干预均1次/d,连续7 d。干预前后对各组大鼠进行Zea-Longa评分、神经功能缺损评分(mNSS)及神经行为学评分。TTC染色法测定大鼠脑梗死体积百分比;免疫组织化学法检测大鼠缺血大脑皮层区IL-1β、IL-6、肿瘤坏死因子-α(TNF-α)、IL-4、IL-10、转化生长因子-β(TGF-β)的表达;免疫荧光双标法检测大鼠缺血大脑皮层区小胶质细胞标志物CD16、CD206的表达;Western blot法检测大鼠缺血大脑皮层区PPARγ、p-NF-κB p65、诱导型一氧化氮合酶(iNOS)和精氨酸酶1(ARG1)的蛋白表达水平。结果:与假手术组比较,模型组大鼠Zea-Longa评分、mNSS评分和神经行为学评分升高(P<0.01),脑梗死体积增大(P<0.01),缺血大脑皮层区p-NF-κB p65/NF-κB p65比值、iNOS、CD16、IL-1β、IL-6及TNF-α表达升高(P<0.01),PPARγ、ARG1、CD206、IL-4、IL-10及TGF-β表达降低(P<0.01)。与模型组比较,电头针组大鼠Zea-Longa评分、mNSS评分和神经行为学评分降低(P<0.01),脑梗死体积缩小(P<0.01),缺血大脑皮层区p-NF-κB p65/NF-κB p65比值、iNOS、CD16、IL-1β、IL-6及TNF-α表达降低(P<0.01),PPARγ、ARG1、CD206、IL-4、IL-10及TGF-β表达升高(P<0.01);与电头针组比较,电头针+抑制剂组大鼠Zea-Longa评分、mNSS评分和神经行为学评分升高(P<0.01),脑梗死体积增大(P<0.01),p-NF-κB p65/NF-κB p65比值、iNOS、CD16、IL-1β、IL-6及TNF-α表达升高(P<0.01),PPARγ、ARG1、CD206、IL-4、IL-10及TGF-β表达降低(P<0.01)。结论:电头针可减轻MCAO大鼠炎性损伤,改善神经功能缺损程度,缩小脑梗死体积,其机制可能与其调控PPARγ/NF-κB信号通路诱导小胶质细胞向M2表型转化有关。
Objective To observe the effect of electroacupuncture(EA)of scalp acupoints on peroxisome proliferator-activated receptorγ(PPARγ),phosphorylated nuclear factor kappa-B p65(p-NF-κB p65),interleukin(IL)-1βand IL-4 as well as CD16 and CD206 in the ischemic cortical tissues of rats with ischemic stroke(IS),so as to explore its mechanisms underlying alleviation of inflammatory damage in ischemic stroke.Methods Male SD rats were randomly assigned to shamsurgery,model,EA,and EA+inhibitor groups,with 12 rats in each group.The IS model was established by occlusion of the middle cerebral artery with suture-embolus in reference with the modified Zea-Longa method.EA(2 Hz/100 Hz,1 mA)was applied to bilateral“Dingnie Qianxiexian”(MS6)for 30 min,once a day for 7 consecutive days.Rats of the EA+inhibitor group received intraperitoneal injection of GW9662 solution(10 mg/mL,an inhibitor of PPARγ).The neurological deficit severity(0-3 points)was assessed using Zea-Longa score,modified neurological severity score(mNSS)and neurobehavioral score before and after the intervention.The cerebral infarction volume was determined using 2,3,5-triphenyltetrazoliumchloride(TTC)staining,and the expression of IL-1β,IL-6,tumor necrosis factor-α(TNF-α),IL-4,IL-10,and transforming growth factor-β(TGF-β)in the ischemic cortex was detected using immunohistochemistry.The immunofluorescence double labeling of microglial markers CD16 and CD206in the ischemic cortex was observed.Western blot was used to detect the expression of PPARγ,p-NF-κB p65,inducible nitric oxide synthase(iNOS),and arginase1(ARG1)proteins in the ischemic cortex.Results Compared with the shamsurgery group,the Zea-Longa score,mNSS score and neurobehavioral score,cerebral infarction volume,and the expression levels of p-NF-κB p65,iNOS,the number of CD16/IBA1 double-labelled cells,and the immunoactivity of IL-1β,IL-6 and TNF-αwere significantly increased(P<0.01),while the expression of PPARγand ARG1 proteins,number of CD206/IBA1 double-labelled cells,and the immunoactivity of IL-4,IL-10 and TGF-βwere significantly decreased(P<0.01)in the model group.After the intervention,in comparison with the model group,the Zea-Longa score,mNSS score and neurobehavioral score,the cerebral infarction volume,number of CD16/IBA1double-labelled cells,and the immunoactivity of IL-1β,IL-6 and TNF-α,and the expression levels of p-NF-κB p65,iNOS,number of CD16/IBA1 double-labelled cells,and the immunoactivity of IL-1β,IL-6 and TNF-αwere considerably decreased(P<0.01),whereas the expression levels of PPARγand ARG1 proteins,the number of CD206/IBA1 double-labelled cells,and the immunoactivity of IL-4,IL-10 and TGF-βwere strikingly increased(P<0.01)in the EA group.After administration of GW9662,the effects of EA were eliminated in reducing the Zea-Longa score,mNSS score and neurobehavioral score,in downregulating the expression of iNOS,p-NF-κB p65,IL-1βand TNF-α(P<0.01),and in upregulating the expression of ARG-1,PPARy,and IL-4,and the number of CD206/IBA1 double-labelled cells(P<0.01)in the EA+inhibitor group.Conclusion EA of MS6 can alleviate the degree of neurological deficit and reduce the volume of cerebral infarction in IS rats,which may be related to its functions in regulating the PPARγ/NF-κB signaling pathway and in promoting the transformation of microglia towards M2 phenotype.
作者
包伟伟
罗文君
张小强
彭晓云
李淑芳
李兴兰
张延菊
田甜
朱玲桂
王金海
BAO Wei-wei;LUO Wen-jun;ZHANG Xiao-qiang;PENG Xiao-yun;LI Shu-fang;LI Xing-lan;ZHANG yan-ju;TIAN Tian;ZHU Ling-gui;WANG Jin-hai(Gansu University of Chinese Medicine,Lanzhou 730000,China;Department of Traditional Chinese Medicine,The Second Hospital of Lanzhou University,Lanzhou 730030;Department of Rehabilitation,The Second Hospital of Lanzhou University,Lanzhou 730030;Department of Chest Surgery,The Second Hospital of Lanzhou University,Lanzhou 730030)
出处
《针刺研究》
北大核心
2025年第10期1152-1160,共9页
Acupuncture Research
基金
国家自然科学基金项目(No.82260962、81960896)
甘肃自然科学基金项目(No.22JR11RA064)
甘肃省高等学校创新基金项目(No.2021A-210)。
