二十五味珊瑚丸调控Akt/mTOR/GSK-3β信号通路改善阿尔茨海默病小鼠的作用机制

Ershiwuwei Shanhu Pills regulate Akt/mTOR/GSK-3βsignaling pathway to alleviate Alzheimer′s disease mice

该研究基于Akt/mTOR/GSK-3β信号通路研究藏族药二十五味珊瑚丸(Ershiwuwei Shanhu Pills,ESP)对阿尔茨海默病(Alzheimer′s disease,AD)小鼠的作用机制。将BALB/c小鼠随机分为空白组,模型组,二十五味珊瑚丸低(200 mg·kg^(-1))、中(400 mg·kg^(-1))、高(800 mg·kg^(-1))剂量组和盐酸多奈哌齐(donepezil hydrochloride,DP HCL)组。除空白组外,其他各组灌胃20 mg·kg^(-1)三氯化铝并腹腔注射120 mg·kg^(-1)D-半乳糖,连续56 d构建阿尔茨海默病模型并给予药物治疗。采用Morris水迷宫法检测小鼠学习记忆能力,检测小鼠海马体中p-tau蛋白含量,检测小鼠海马体和血清超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)和总抗氧化能力(T-AOC)水平,苏木精-伊红染色(HE)和尼氏染色对小鼠全脑进行病理学观察,TUNEL染色观察小鼠皮层区的细胞凋亡情况。Western blot法测定小鼠海马体的p-mTOR、p-Akt和GSK-3β蛋白表达。结果显示,与模型组相比,二十五味珊瑚丸给药组小鼠的潜伏期和找到原平台的路径显著缩短,全脑病理损伤程度显著改善;TUNEL阳性细胞表达有所减少;海马体中p-tau蛋白含量显著降低;海马体和血清中SOD、CAT、T-AOC水平显著升高,MDA水平显著降低;海马体p-mTOR和p-Akt蛋白表达显著增加,GSK-3β蛋白表达显著减少。研究结果表明,二十五味珊瑚丸可减轻D-半乳糖联合三氯化铝致小鼠阿尔茨海默病引起的学习记忆能力衰退和氧化损伤,其作用机制可能与Akt/mTOR/GSK-3β信号通路有关。

The present study investigated the mechanism of the Tibetan patent medicine Ershiwuwei Shanhu Pills(ESP)in alleviating Alzheimer′s disease in mice via Akt/mTOR/GSK-3βsignaling pathway.BALB/c mice were randomly assigned into a blank control group,a model group,low(200 mg·kg^(-1)),medium(400 mg·kg^(-1))and high(800 mg·kg^(-1))dose groups of ESP,and donepezil hydrochloride group.Except the blank control group,the other groups were given 20 mg·kg^(-1)aluminum chloride by gavage and 120 mg·kg^(-1)D-galactose by intraperitoneal injection for 56 days to establish Alzheimer′s disease model.Morris water maze was used to detect the learning and memory ability of mice.The level of p-tau protein in mouse hippocampus and the levels of superoxide dismutase(SOD),malondialdehyde(MDA),catalase(CAT),and total antioxidant capacity(T-AOC)in hippocampus and serum were detected.Hematoxylin-eosin staining and Nissl staining were performed for the pathological observation of whole brain in mice.TdT-mediated dUTP nick-end labeling(TUNEL)staining was employed for the observation of apoptosis in mouse cortex.Western blot was adopted to detect the protein levels of p-mTOR,p-Akt,and GSK-3βin the hippocampus.Compared with the model group,the ESP groups showcased alleviated pathological damage of the whole brain,decreased TUNEL positive cells,reduced level of p-tau protein in hippocampus,and risen SOD,CAT,and T-AOC levels and declined MDA level in hippocampus and serum.Furthermore,the ESP groups had up-regulated protein levels of p-mTOR and p-Akt while down-regulated protein level of GSK-3βin hippocampus.Therefore,ESP can alleviate the learning and memory decline and oxidative damage in mice with Alzheimer′s disease induced by D-galactose combined with aluminum chloride,which may be related to Akt/mTOR/GSK-3βsignaling pathway.