枸杞叶改善D-半乳糖致亚急性衰老小鼠学习与记忆能力的效应部位与作用机制研究  

Researches on Effective Fraction and Mechanism of Lycium Barbarum Leaves on Improving Learning and Memory Abilities of D-Galactose-Induced Subacute Aging Mice

作　　者：仝佳祥 陈杨 李璇 朱梓强 宿树兰[1] 郭盛[1] 康宏杰 段金廒[1] 朱悦[1] Tong Jiaxiang;Chen Yang;Li Xuan;Zhu Ziqiang;Su Shulan;Guo Sheng;Kang Hongjie;Duan Jin'ao;Zhu Yue(School of Pharmacy,Nanjing University of Chinese Medicine,Jiangsu Key Laboratory for Traditional Chinese Medicine Formulae Research,Jiangsu Key Laboratory for High Technology Research of TCM Formulae,Nation and Local Union Project Research Center of Chinese Medicinal Resources Industrialization and Innovative Drug from TCM Formulae and Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization,Nanjing 210023,China;Ningxia Innovation Center of Wolfberry,Yinchuan 750000,China)

机构地区：[1]南京中医药大学药学院,江苏省方剂研究重点实验室/江苏省方剂高技术研究重点实验室/中药资源产业化与方剂创新药物国家地方联合工程研究中心/江苏省中药资源产业化过程协同创新中心,南京210023 [2]宁夏枸杞创新中心,银川750000

出　　处：《世界科学技术-中医药现代化》2024年第1期48-60,共13页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：宁夏回族自治区科学技术厅重点研发计划(2022BBF02004):枸杞叶新资源药材及其新药制剂创制,负责人:康宏杰;宁夏回族自治区科学技术厅重点研发计划(2021BEF02009):枸杞子组方防治阿尔茨海默症组分配伍保健产品创制,负责人:朱悦,郭盛;宁夏回族自治区科学技术厅科技基础条件建设项目(2021DPC05005):枸杞创新中心基础条件建设,负责人:段金廒

摘　　要：目的研究枸杞叶改善D-半乳糖致亚急性衰老小鼠学习与记忆能力的效应部位与作用机制。方法建立D-半乳糖致亚急性衰老小鼠模型,制备枸杞叶不同溶剂提取部位,通过Y迷宫实验和新物体识别实验评价枸杞叶不同溶剂提取部位对模型小鼠学习与记忆能力的影响;苏木素-伊红(HE)以及尼氏(Nissl)染色观察小鼠海马区病理形态学变化;酶联免疫吸附法检测小鼠海马组织脑源性神经营养因子(Brain-derived neurotrophic factor,BDNF)、神经生长因子(Nerve growth factor,NGF)、胶质细胞源性神经营养因子(Glial cell line-derived neurotrophic factor,GDNF)以及肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、γ干扰素(IFN-γ)、白细胞介素10(IL-10)的含量;试剂盒检测小鼠海马组织中超氧化物歧化酶(SOD)的活力,丙二醛(MDA)、谷胱甘肽(GSH)的含量;TUNEL法检测小鼠脑组织海马区细胞凋亡情况;蛋白印迹法检测小鼠海马组织抗氧化蛋白Nrf2、HO-1以及凋亡蛋白Caspase-3、Caspase-9的表达。结果枸杞叶水提部位和80%醇沉上清部位显著改善模型小鼠学习与记忆能力,改善小鼠海马区组织病理损伤,增加小鼠海马区尼氏体数目,并促进神经营养因子BDNF、NGF、GDNF的表达;下调致炎因子TNF-α、IL-1β、IFN-γ的表达,上调抗炎因子IL-10表达;提升SOD活力和GSH表达,降低MDA表达;提高Nrf2、HO-1抗氧化蛋白的表达;降低Caspase-3、Caspase-9凋亡通路蛋白的表达;抑制模型小鼠脑组织海马区细胞凋亡。结论枸杞叶水提部位和80%醇沉上清部位是枸杞叶改善D-半乳糖致亚急性衰老小鼠学习与记忆能力的效应部位,其作用途径与抑制中枢氧化应激与炎性应激导致的神经元凋亡有关。Objective To study the effective fraction and mechanism of Lycium barbarum leaves on improving learning and memory ability of subacute aging mice induced by D-galactose injection.Methods The model of subacute aging mice was developed by injection of D-galactose subcutaneously,and different extracts of Lycium barbarum leaves were prepared.The effects of the extracts of Lycium barbarum leaves on the learning and memory ability of model mice were evaluated by Y maze experiment and new object recognition experiment.The pathomorphological changes of hippocampus in mice were observed by hematoxylin-eosin and Nissl staining.The levels of brain-derived neurotrophic factor(BDNF),nerve growth factor(NGF),glial cell line-derived neurotrophic factor(GDNF),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interferon-γ(IFN-γ)and interleukin-10(IL-10)in hippocampus of mice were detected by enzyme-linked immunosorbent assay.The activities of superoxide dismutase(SOD)and the contents of glutathione(GSH)and malondialdehyde(MDA)in hippocampus of mice were detected by related assay kits.Detection of apoptosis in the hippocampal region of mouse brain tissue using the TUNEL method.Western blotting analysis was used to detect the expressions of antioxidant proteins Nrf2,HO-1 and apoptotic proteins Caspase-3,Caspase-9 in hippocampus of mice.Results The water extraction part and 80%alcohol precipitation supernatant part of Lycium barbarum leaves significantly improved the learning and memory ability of model mice,improved the pathological damage of hippocampus in mice,increased the number of Nissl bodies in hippocampus of mice,and promoted the expression of neurotrophic factors BDNF,NGF and GDNF,and promoted the expression of neurotrophic factors BDNF,NGF and GDNF.Pro-inflammatory factors TNF-α,IL-1βand IFN-γexpression declines while anti-inflammatory factor IL-10 expression rises.The activity of SOD and the expression of GSH were increased,and the expression of MDA was decreased.Increase the expression of Nrf2 and HO-1 antio

关 键 词：枸杞叶 衰老 认知损伤 凋亡 

分 类 号：R285.5[医药卫生—中药学]