梓醇对大脑皮质神经元的抗衰老保护作用研究  

Study of anti-aging protective effect of catalpol on cultured cortical neurons from rats

作　　者：王静欢 刘珂[1] 万东[2] 祝慧凤[1] WANG Jing huan;LIU Ke;WAN Dong;ZHU Hui feng(College of Pharmaceutical Sciences& College of Chinese Medicine, Southwest University, Chongqing400715, China;Dept of Emergency and Critical Care Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing400016, China)

机构地区：[1]西南大学药学院暨中医药学院中药药理学实验室,重庆400715 [2]重庆医科大学附属第一医院急诊医学科&重症医学科,重庆400016

出　　处：《中国药理学通报》2017年第12期1691-1697,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81073084);重庆市自然科学基金资助项目(No CSTC2010BB5127;CSTC2014jcyj A10083);教育部中央高校重点项目(No XDJK2012B010)

摘　　要：目的探讨梓醇对正常皮质神经元的抗衰老作用及其可能机制。方法原代培养新生24 h SD大鼠皮质神经元,分为正常组、梓醇低(0.1 mg·L^(-1))、中(1 mg·L^(-1))、高(10 mg·L^(-1))剂量组。上述培养基中的神经元培养至13 d,显微观察7~13 d的各组细胞形态;MTT法检测7~13 d各组细胞的细胞活力;免疫荧光分别检测13 d各组神经元pS6和Map-2表达以显示细胞的内在活性和轴突生长情况;Western blot法检测13 d各组细胞GAP-43、p-S6、PI3K、pPI3K、Akt、p-Akt、m TOR、p-m TOR蛋白的表达。结果与正常组相比,梓醇给药各组均能延缓神经元的衰老,促进神经元的存活,提高神经元的细胞活力(P<0.05);诱导神经元内在活性,促进p-S6阳性细胞数增加(P<0.05);促进轴突生长(P<0.05)。且不同浓度的梓醇均能促进13 d的神经元GAP-43、p-S6、PI3K、p-PI3K、Akt、p-Akt、m TOR、p-m TOR蛋白表达增加(P<0.05)。结论梓醇具有抗大脑皮质神经元衰老作用,其机制可能与调控PI3K/Akt/m TOR信号通路,增强神经元生长活性有关。AimTo investigate the anti aging effect of catalpol on normal cortical neurons and its possible mechanism.MethodsPrimary cultured cortical neurons from24h newly born rats were dissociated and cultured.The cells were divided into normal group,catalpol(0.1mg·L-1)group,catalpol(1mg·L-1)group and catalpol(10mg·L-1)group.Neurons were cultured for13d,and the cell morphology was observed by microscope.Cell activity was detected by MTT assay.p S6and Map2of13d neurons were detected by immunofluorescence to reveal cell viability and the axon growth.GAP43,p S6,PI3K,p PI3K,Akt,p Akt,mTOR and p mTOR protein expression of13d neurons were detected by Western blot.ResultsCompared with normal group,different concentrations of catalpol could delay the aging of neurons,promote their survival and increase cell viability.The nμmber of p S6positive cells increased.Different concentrations of catalpol could promote axon growth.GAP43,p S6,PI3K,p PI3K,Akt,p Akt,mTOR and p mTOR protein expression of13d neurons increased.ConclusionsCatalpol has anti aging effects on cerebral cortical neurons,and its mechanism may be related to regulating PI3K/Akt/mTOR signaling pathway and enhancing neuronal growth activity.

关 键 词：梓醇 皮质神经元 抗衰老 细胞活性 轴突生长 PI3K/AKT/MTOR 

分 类 号：R-332[医药卫生] R284.1