摘要
目的观察电针“百会”“水沟”穴对脑缺血再灌注损伤大鼠及神经细胞自噬的影响。方法SD雄性大鼠随机分为假手术组、模型组、干预组各16只。假手术组麻醉后只分离左侧颈总、颈内、颈外动脉,不结扎、插线。模型组、干预组采用Zea Longa线栓法制备大鼠大脑中动脉缺血再灌注模型。干预组电针“百会”“水沟”,电流强度1 mA,疏波频率3 Hz,密波频率15 Hz,1次/d,20 min/次,连续5 d。假手术组和模型组均不予电针处理,其他处理同干预组。制备模型5 d后,用Longa神经功能评分法对各组进行神经功能评分,2,3,5-氯化三苯基四氮唑(TTC)染色法测定脑梗死体积,酶联免疫吸附试验(ELISA)检测脑脊液白细胞介素(IL)-6、IL-18、肿瘤坏死因子(TNF)-α的水平,实时荧光定量聚合酶链反应(RT-qPCR)和Western印迹分别检测缺血区海马组织自噬相关蛋白磷脂酰肌醇-3激酶(PI3K)、蛋白激酶B(Akt)、哺乳动物雷帕霉素靶蛋白(mTOR)mRNA和蛋白表达。结果与假手术组相比,模型组神经功能评分明显升高,脑梗体积明显增加,脑脊液IL-6、IL-18、TNF-α水平明显升高,PI3K、Akt、mTOR mRNA及蛋白表达明显降低(均P<0.05);与模型组相比,干预组神经功能评分明显降低,脑梗体积明显减小,脑脊液IL-6、IL-18、TNF-α水平明显降低,PI3K、Akt、mTOR mRNA及PI3K、Akt蛋白表达明显增高(均P<0.05)。结论电针“百会”“水沟”穴可能通过激活PI3K/Akt/mTOR信号通路,抑制缺血区细胞自噬从而改善脑缺血再灌注损伤大鼠的神经功能,减少脑梗死体积。
Objective To observe the effect of electroacupuncture at"Baihui"and"Shuigou"acupoints on cerebral ischemiareperfusion injury and autophagy of nerve cells in rats.Methods SD male rats were randomly divided into sham operation group,model group and intervention group,with 16 rats in each group.After anesthesia,only the left common carotid artery,internal carotid artery and external carotid artery were separated in the sham operation group,without ligation or insertion.The rat middle cerebral artery ischemia-reperfusion model was established by Zea Longa thread occlusion method in model group and intervention group.The intervention group was treated with electroacupuncture"Baihui"and"Shuigou",current intensity 1mA,the sparse wave frequency was 3 Hz,and the dense wave frequency was 15 Hz,once a day,20 min each time for 5 consecutive days.The sham operation group and the model group were not treated with electroacupuncture,and the other treatments were the same as those in the intervention group.Five days after the establishment of the model,the neurological function in each group was evaluated by Longa neural function score,the volume of cerebral infarction was measured by 2,3,5-Triphenyte-trazoliumchloride(TTC)staining,the levels of interleukin(IL)-6,IL-18 and tumor necrosis factor(TNF)-αin cerebrospinal fluid were detected by enzyme-linked immuno sorbent assay(ELISA),and the expressions of autophagy-related proteins phosphatidylinositol-3-kinase(PI3K),protein kinase B(Akt),mammalian target of rapamycin(mTOR)mRNA and protein in hippocampal tissues of ischemic area were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot respectively.Results Compared with the sham operation group,the neurological function score of the model group was significantly increased,the volume of cerebral infarction was significantly increased,the levels of IL-6,IL-18 and TNF-αin cerebrospinal fluid were significantly increased,the expressions of PI3K,Akt and mTOR mRNA and protein were significantly decreased(all P<0.05).Compared with the model group,the neurological function score of the intervention group was significantly decreased,the volume of cerebral infarction was significantly decreased,the levels of IL-6,IL-18 and TNF-αin cerebrospinal fluid were significantly decreased,the expressions of PI3K,Akt and mTOR mRNA and protein were significantly increased(all P<0.05).Conclusions Electroacupuncture at"Baihui"and"Shuigou"acupoints might improve the neurological function of rats with cerebral ischemia-reperfusion injury and reduce the volume of cerebral infarction by activating PI3K/Akt/mTOR signal pathway and inhibiting autophagy in ischemic zone.
作者
黄夏荣
封蔚彬
胡莉芝
谭金曲
周桂娟
王金玲
尹林伟
周君
刘红雅
曾亚华
HUANG Xia-Rong;FENG Wei-Bin;HU Li-Zhi(The First Affiliated Hospital of Hengyang Medical School,University of South China,Hengyang 421001,Hunan,China)
出处
《中国老年学杂志》
CAS
北大核心
2024年第10期2443-2448,共6页
Chinese Journal of Gerontology
基金
国家自然科学基金(81973917)
湖南省自然科学基金(2021JJ30616,2021JJ30617,2020JJ5512)
湖南省临床医疗技术创新引导项目(2020SK51811)
湖南省卫生健康委项目(20200474)
关键词
电针
百会
水沟
脑缺血再灌注损伤
自噬
Electroacupuncture
Baihui
Shuigou
Cerebral ischemia-reperfusion injury
Autophagy
