针刺通过调控PI3K/Akt/mTOR通路改善血管性痴呆大鼠认知功能的机制研究  

Acupuncture improves cognitive function of vascular dementia rats by regulating PI3K/Akt/mTOR pathway

作　　者：李龙春 李晓晔[2] 杜小华 LI Long-chun;LI Xiao-ye;DU Xiao-hua(Clinical School of Acupuncture and Rehabilitation,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Department of Rehabilitation,Baoan District Maternal and Child Health Hospital of Shenzhen City,Shenzhen 518101,Guangdong Province;College o£Acupuncture and Massage,Guangxi University of Chinese Medicine,Nanning 530001)

机构地区：[1]广州中医药大学针灸康复临床医学院,广州510405 [2]深圳市宝安区妇幼保健院康复科,广东深圳518101 [3]广西中医药大学针灸推拿学院,南宁530001

出　　处：《针刺研究》2021年第10期851-856,共6页Acupuncture Research

基　　金：广东省医学科学技术研究基金项目(No.A2018179)。

摘　　要：目的:探讨针刺通过调控磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)通路改善血管性痴呆(VD)大鼠认知功能的机制。方法:Wistar大鼠随机分为假手术组、模型组、非穴组和针刺组,每组18只。采用永久性双侧颈动脉结扎法制备VD大鼠模型。针刺组针刺"百会"和双侧"足三里",非穴组针刺两个固定非穴点,均每天1次,针刺6 d,休息1 d,连续2周。Morris水迷宫实验检测各组大鼠学习和空间记忆能力;ELISA法检测各组大鼠海马组织中活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量以及乙酰胆碱脂酶(AChE)、乙酰胆碱转移酶(ChAT)活力;JC-1法检测各组大鼠海马组织线粒体膜电位(MMP)的水平;Western blot法检测各组大鼠海马组织中PI3K、p-Akt及mTOR表达水平。结果:与假手术组相比,模型组大鼠逃避潜伏期延长(P<0.05),ROS和MDA含量以及AChE活力显著升高(P<0.05),西南象限停留时间缩短(P<0.05),SOD活性、ChAT活力、MMP以及PI3K、p-Akt和mTOR蛋白表达水平显著降低(P<0.05)。与模型组和非穴组相比,针刺组大鼠逃避潜伏期缩短(P<0.05),ROS和MDA含量以及AChE活力显著降低(P<0.05),西南象限停留时间延长(P<0.05),SOD活性、ChAT活力、MMP以及PI3K、p-Akt和mTOR蛋白表达水平显著升高(P<0.05)。与模型组相比,非穴组大鼠各项指标差异无统计学意义(P>0.05)。结论:针刺可能通过激活PI3K/Akt/mTOR信号通路,缓解VD大鼠海马组织的氧化应激及线粒体膜电位降低,提高VD大鼠的认知功能。Objective To explore the mechanism of acupuncture in improving cognitive ability by regulating hippocampal phosphatidylinositol-3 kinase(PI3 K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) pathway in vascular dementia(VD) rats.Methods A total of 80 male SD rats were randomized into sham operation, model, non-acupoint and acupoint groups(n=18 per group). The VD model was established by ligation of the bilateral common carotid arteries. For rats of the acupoint group, "Baihui"(GV20) and bilateral "Zusanli "(ST36) were needled and stimulated by twirling the needles with reinforcing method, and for rats of the non-acupoint group, the bilateral subcostal spots(about 10 mm superior to the iliac cresta) were needled and stimulated by twirling the needles with uniform reinforcing and reducing method. The treatment was conducted once daily, 6 times a week for two weeks, beginning 3 days after successful modeling. Rats of the sham operation group and model group received grasps as those in the acupoint groups. Morris water maze test was used to detect the abilities of learning and spatial memory. The contents of reactive oxygen species(ROS), malondialdehyde(MDA), superoxide dismutase(SOD) and the activities of acetylcholinesterase(AChE) and acetylcholine transferase(ChAT) in the hippocampus tissue were detected by using ELISA, changes of hippocampal mitochondrial membrane potential(MMP) detected using JC-1 fluorescence probe, and the expression levels of hippocampal phosphorylated(p)-PI3 K, p-Akt and mTOR proteins measured using Western blot. Results Compared with the sham operation group, the escape latency, contents of ROS and MDA, and AChE activity were significantly increased(P<0.05), and the spatial memory ability, SOD activity, ChAT activity, MMP, p-PI3 K, p-Akt and mTOR expression levels were significantly decreased in the model group(P<0.05). Compared with the model group, carotid artery ligature-induced increase of the escape latency, contents of ROS and MDA, and AChE activity, and decrease of the spa

关 键 词：针刺 血管性痴呆 认知功能 海马 氧化应激 线粒体膜电位 PI3K/Akt/mTOR信号通路 

分 类 号：R246.6[医药卫生—针灸推拿学]