摘要
【目的】探讨马尾松松针提取物(PNE)对D-半乳糖致衰小鼠大脑神经元的改善作用。【方法】将50只健康小鼠分成对照组、模型组、PNE低剂量组、PNE中剂量组和PNE高剂量组,腹腔注射D-半乳糖建立衰老小鼠模型,造模同时用PNE(200 mg/kg、400 mg/kg、800 mg/kg)进行灌胃干预。食饵迷宫观察小鼠行为学变化,生物显微技术观察大脑皮层结构的变化,比色法检测大脑皮层胆碱乙酰基转移酶(ChAT)、乙酰胆碱酯酶(AChE)、(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化酶(GPx)活性及丙二醛(MDA)含量,酶联免疫吸附(ELISA)法测定大脑皮层白细胞介素1(IL-1β)和肿瘤坏死因子(TNF-α)含量,Western-blotting检测大脑皮层中磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)/mTOR、磷酸化腺苷酸活化蛋白激酶(p-AMPK)/AMPK、核转录因子红系2相关因子2(Nrf2)、Kelch样环氧氯丙烷相关蛋白1(KKeap-1)、血红素加氧酶1(HO-1)表达。【结果】PNE使衰老小鼠食饵迷宫试验的错误次数及总测试时间明显降低(P<0.05),大脑皮层病理变化得到改善;大脑皮层SOD、CAT、GPx、ChAT活性显著升高,IL-1β、TNF-α和MDA含量及AChE活性显著降低(P<0.05)。此外,PNE干预后,小鼠大脑皮层Nrf2、HO-1、p-AMPK/AMPK表达显著增加(P<0.05),Keap-1、p-mTOR/mTOR表达显著减少(P<0.05)。【结论】PNE能够改善D-半乳糖致衰小鼠大脑神经元损伤并提高小鼠的学习记忆能力,其机制可能与抗氧化、抑制炎症因子分泌、调节AMPK/mTOR和Nrf2/HO-1信号通路有关。
【Objective】To study the effect of Pinus massoniana needle extracts(PNE)on cerebral neurons in D-galactose(D-gal)-induced aging mice.【Method】Fifty mice were divided into five groups:control group,model group,low-dose PNE group,medium-dose PNE group and high-dose PNE group.The aging mouse model was established by intraperitoneal injection of D-galactose and the mice were given PNE at doses of 200 mg/kg,400 mg/kg or 800 mg/kg by intragastric administration during the modelling period.Behavioral changes in the mice were observed using a bait maze.At the same time,the step-down test was performed to determine the learning and memory ability of the mice.The cerebral cortex was observed by biomicroscopy.Spectrophotometry was used to determine the activities of choline acetyltransferase(ChAT),acetylcholinesterase(AChE),superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GPx)and malondialdehyde(MDA)levels in the cerebral cortex.And the levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the cerebral cortex were examined by ELISA.Western blotting was used to detect the age-related proteins pro-mammalian target of rapamycin(p-mTOR)/mTOR,proactivated protein kinase(p-AMPK)/AMPK,nuclear factor erythroid 2-related factor 2(Nrf2),Kelch-like epichlorohydrin-related protein 1(Keap-1)and haem oxygenase1(HO-1)in the cerebral cortex.【Result】After PNE treatment,the error frequency and total test time of the bait maze were significantly lower than those of the D-gal group(P<0.05),and the pathological changes of the cerebrum were significantly im⁃proved.The activities of cerebral SOD,CAT,GPx,ChAT of the PNE dose group were tremendously higher than those of the model group(P<0.05).The levels of IL-1β,TNF-αand MDA and AChE activity were significantly lower than those in the model group(P<0.05).In addition,PNE significantly increased the expression of Nrf2,HO-1 and p-AMPK/AMPK(P<0.05)and decreased the expression of Keap-1 and p-mTOR/mTOR in the cerebral cortex(P<0.05).【Conclusion】PNE could effectively alleviate D-galinduced cerebral neuronal damage in aging mice and improve the learning and memory abilities of mice,which may be due in part to an increase in the activities of antioxidant enzymes,a reduction in the secretion of inflammatory cytokines,and regulation of the AMPK/mTOR and Nrf2/HO-1 signaling pathways.
作者
何九军
陈文东
王昱
HE Jiujun;CHEN Wendong;WANG Yu(College of Agriculture and Forestry,Longnan Normal University,Chengxian 742500,China;Center for Research&Development of Longnan Characteristic Agro-bioresources,Longnan Normal Vnirersity,Chengxian 742500,China)
出处
《甘肃农业大学学报》
北大核心
2025年第1期27-34,共8页
Journal of Gansu Agricultural University
基金
甘肃省科技重大专项(21ZD4NK045)
甘肃省高等学校科研创新基金项目(2020A-276)
甘肃省陇南市社会化出资科技计划项目(2021-SZ-08)。
作者简介
第一作者:何九军,副教授,主要从事天然产物药理研究。E-mail:1216600861@qq.com;通信作者:王昱,教授,主要从事天然产物药理、细胞与发育生物学研究。E-mail:gswangyu@126.com。
