电针调控PI3K/AKT/mTOR通路改善血管性痴呆大鼠海马神经元炎性和氧化损伤机制  

Mechanism of electroacupuncture in improving hippocampal neuronal inflammatory and oxidative injuries in vascular dementia rats by regulating PI3K/AKT/mTOR pathway

作　　者：毕珂瑶 李赫妍 苏景超 陶树琴 唐巍[1] BI Keyao;LI Heyan;SU Jingchao;TAO Shuqin;TANG Wei(Anhui University of Chinese Medicine,Hefei 230038,China;Institute of Acupuncture and Meridians,Anhui University of Chinese Medicine,Hefei 230012,China)

机构地区：[1]安徽中医药大学,合肥230038 [2]安徽中医药大学针灸经络研究所,合肥230012

出　　处：《上海针灸杂志》2024年第3期326-334,共9页Shanghai Journal of Acupuncture and Moxibustion

基　　金：安徽中医药大学校级探索性科研项目(2021zxts32);2021年度高校科学研究项目(YJS20210481);安徽省自然科学基金面上项目(2108085MH309);省级研究生学术创新项目(2022xscx098)。

摘　　要：目的 探讨电针百会、足三里穴调控磷脂酰肌醇3激酶(phosphatidylinositol 3 kinase, PI3K)/蛋白激酶B(protein kinase B, AKT)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin, mTOR)通路改善血管性痴呆(vascular dementia, VD)大鼠海马组织神经元炎性和氧化损伤机制。方法 45只雄性SD大鼠,随机分为假手术组、模型组和电针组,每组15只。模型组与电针组大鼠采用双侧颈动脉结扎法(2-VO)制备VD模型,假手术组颈动脉分离不结扎。电针组造模3 d后电针百会、足三里穴,每日1次,连续2周,第7天和第14天间断治疗。Morris水迷宫实验检测各组大鼠空间记忆和学习能力。Nissl染色法观察海马区神经元结构形态;酶联免疫吸附测定法检测血清白介素-1β(interleukin-1β, IL-1β)、IL-6(interleukin 6, IL-6)以及谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)含量;生化检测海马组织丙二醛(malonaldehyde, MDA)和超氧化物歧化酶(superoxide dismutase, SOD)含量;Western blot法检测海马组织中p-PI3K、p-Akt及p-mTOR蛋白表达量;免疫组化法观察海马组织mTOR蛋白阳性表达。结果 与假手术组比较,模型组大鼠空间记忆和学习能力明显下降(P<0.01),Nissl染色显示神经元受损明显,MDA、IL-1β和IL-6含量显著升高(P<0.01),GSH-Px含量、SOD含量、p-PI3K、p-Akt及p-mTOR蛋白表达显著降低(P<0.01),mTOR阳性表达量降低(P<0.01)。与模型组比较,电针组大鼠空间记忆和学习能力显著提升(P<0.01),Nissl染色显示神经元形态显著改善,MDA、IL-1β和IL-6含量显著降低(P<0.01),GSH-Px含量、SOD含量、p-PI3K、p-Akt及p-mTOR蛋白表达水平显著升高(P<0.01), mTOR阳性表达量明显升高(P<0.01)。结论 电针可能通过上调PI3K/AKT/mTOR信号通路来降低炎症反应与氧化损伤,从而保护VD大鼠海马神经元,并改善其认知能力。Objective To discuss the mechanism of electroacupuncture(EA)at Baihui(GV20)and Zusanli(ST36)in improving hippocampal neuronal inflammatory and oxidative injuries in vascular dementia(VD)rats by regulating the phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway.Method Forty-five male Sprague-Dawley(SD)rats were randomly allocated to a sham operation group,a model group,and an EA group,with 15 rats in each group.Rats in the model and EA groups underwent VD modeling of bilateral carotid artery ligation(2-VO),while rats in the sham operation group only received separation of the carotid artery but without ligation.The EA group was offered EA at Baihui and Zusanli 3 d after modeling,once daily for 2 consecutive weeks with intervals at the 7th and 14th days.The Morris water maze test was used to assess the spatial memory and learning ability of each group of rats.The structure of hippocampal neurons was observed using Nissl staining;the enzyme-linked immunosorbent assay method was adopted to detect the contents of serum interleukin-1b(IL-1b)and interleukin-6(IL-6)and the activity of glutathione peroxidase(GSH-Px);the contents of malondialdehyde(MDA)and superoxide dismutase(SOD)in the hippocampal tissue were determined using the biochemical method;Western blot(WB)was used to detect the protein expression of hippocampal p-PI3K,p-Akt,and p-mTOR;immunohistochemistry was employed to observe the positive expression of hippocampal mTOR protein.Result Compared with the sham operation group,the spatial memory and learning ability declined significantly in the model group rats(P<0.01),along with notable neuronal damage according to Nissl staining,significant increases in the contents of MDA,IL-1b,and IL-6(P<0.01),significant decreases in the contents of GSH-Px and SOD and protein expression of p-PI3K,p-Akt,and p-mTOR(P<0.01),and reduced positive expression of mTOR(P<0.01).Compared with the model group,the EA group presented significant improvements in spatial memory and learning a

关 键 词：电针 血管性痴呆 炎症反应 氧化损伤 PI3K/AKT/mTOR通路 大鼠 

分 类 号：R2-03[医药卫生—中医学]