基于泛素-蛋白酶体系统探讨远志散改善快速老化小鼠学习记忆能力的作用机制  

Mechanism of Yuanzhi Powder on improving the learning and memory ability of SAMP8 mice based on ubiquitin-proteasome system

作　　者：李佳欣 郝彦伟 王少锋 岳胜男 王飞[1] 李斌[1] LI Jia-xin;HAO Yan-wei;WANG Shao-feng;YUE Sheng-nan;WANG Fei;LI Bin(Hospital of Chengdu University of TCM,Chengdu 610072,China)

机构地区：[1]成都中医药大学附属医院,成都610072

出　　处：《中华中医药杂志》2022年第5期2929-2934,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目面上项目(No.82174345),国家自然科学基金青年科学基金项目(No.81704024);中国出土医学文献与文物研究中心科研项目(No.2020C03);四川省科技计划项目(No.2021YJ0435);四川省中医药管理局科学技术研究专项(No.2021MS058)。

摘　　要：目的:探讨远志散调控泛素-蛋白酶体系统(UPS)改善快速老化小鼠(SAMP8)学习记忆能力的作用机制。方法:以PC12细胞为载体观察乳胞素(LAC)对26s蛋白酶体(26s PS)的抑制作用,以6只C57BL/6J小鼠为对象观察双侧海马注射LAC的安全性。18只SAMP8小鼠随机分为模型(MOD)组、盐酸多奈哌齐(DON)组、远志散(YZP)组,12只SAMP8小鼠予双侧海马注射LAC后随机分为乳胞素(LAC)组、乳胞素+远志散(LAC+YZP)组,6只抗快速老化小鼠(SAMR1)为正常对照(CON)组,各组给予相应药物连续灌胃8周。Morris水迷宫实验评价小鼠的学习及记忆能力,HE染色检测海马病理学改变,透射电镜检测海马神经元超微结构变化,Western blot检测海马Ubiquitin、UBE3A、26s PS、UCH-L1、Tau5、pT231-Tau蛋白表达。结果:与MOD组比较,YZP组小鼠逃避潜伏期显著缩短(P<0.05),穿越平台次数显著增加(P<0.05),海马组织病理形态及神经元超微结构病变有所改善,Ubiquitin蛋白表达水平显著下降(P<0.01),UBE3A、26s PS、UCH-L1蛋白表达水平显著升高(P<0.01),pT231-Tau蛋白表达显著下降(P<0.05);与MOD组比较,LAC组小鼠26s PS蛋白表达水平显著下降(P<0.01),pT231-Tau蛋白表达升高(P<0.05);与LAC组比较,LAC+YZP组pT231-Tau蛋白表达下降(P<0.05)。结论:远志散可提高SAMP8小鼠学习和记忆能力,改善海马组织及神经元超微结构病理变化,抑制磷酸化Tau蛋白异常沉积,可能与其调控UPS途径的关键靶点有关。Objective:To investigate the mechanism of Yuanzhi Powder of improving the learning and memory ability of SAMP8 mice via regulating the ubiquitin-proteasome system(UPS).Methods:PC12 cells were used as carriers to observe the inhibitory effect of lactacystin(LAC)on 26s proteasome(26s PS).Six C57BL/6J mice were used to observe the safety of bilateral hippocampal injection of LAC.The 18 SAMP8 mice were randomly divided into MOD group,DON group and YZP group.The 12 SAMP8 mice were randomly divided into LAC group and LAC+YZP group after injection of LAC into bilateral hippocampus.The 6 SAMR1 mice were used as CON group.Each group was given corresponding drugs to gavage treatment for 8 weeks.The learning and memory ability of mice were evaluated by Morris water maze.The pathological changes of hippocampus were observed by HE staining.The ultrastructural changes of hippocampal neurons were observed by transmission electron microscopy.The protein expressions of hippocampal Ubiquitin,UBE3A,26s PS,UCH-L1,Tau5 and pT231-Tau were detected by Western blot.Results:Compared with the MOD group,the escape latency of the YZP group was decreased(P<0.05),while the number of crossing platforms was increased(P<0.05).The pathological changes of hippocampus and ultrastructure of neurons were improved.The protein expression levels of ubiquitin in the hippocampus of YZP group were decreased(P<0.01),the protein expression levels of UBE3A,26s PS and UCH-L1 in the hippocampus were increased(P<0.01),and the protein expression levels of pT231-Tau in the hippocampus were decreased(P<0.05).Compared with the MOD group,the protein expression levels of 26s PS in the hippocampus of LAC group were decreased(P<0.01),and the protein expression levels of pT231-Tau in the hippocampus were increased(P<0.05).Compared with the LAC group,the protein expression levels of pT231-Tau in the hippocampus of LAC+YZP group were decreased(P<0.05).Conclusion:Yuanzhi Powder can improve the learning and memory ability of SAMP8 mice,and rectify the pathological changes of

关 键 词：远志散 阿尔茨海默病 快速老化小鼠 学习记忆能力 泛素-蛋白酶体系统 磷酸化TAU蛋白 乳胞素 海马注射 

分 类 号：R285.5[医药卫生—中药学]