北五味子总木脂素对D-半乳糖诱导的小鼠脑衰老自噬和凋亡的影响  

Effects of Schisandra total lignin on autophagy and apoptosis of mouse brain aging induced by D-galactose

作　　者：于春艳[1] 于春荣 敬舒[3] 李贺[4] 姜恩平[4,5] 鞠文博[1] 陈建光[4] 

机构地区：[1]北华大学基础医学院病理学教研室,吉林吉林132013 [2]北京昭衍新药研究中心股份有限公司毒理部,北京100176 [3]北华大学附属医院普外科,吉林吉林132011 [4]北华大学药学院药理学教研室,吉林吉林132013 [5]广东医学院基础医学院肿瘤研究所,广东东莞523808

出　　处：《吉林大学学报（医学版）》2014年第6期1210-1215,1336,共7页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅自然科学基金资助课题(201215103);吉林省教育厅科研基金资助课题(吉教科合字2012394;2011130;2013194);吉林市科技局科技计划项目资助课题(201233122;201464064);广东医学院博士启动项目资助课题(B2012063)

摘　　要：目的:利用D-半乳糖复制小鼠衰老模型,探讨北五味子总木脂素(SCL)延缓小鼠脑组织衰老的作用及其机制。方法:以50只小鼠为实验对象,实验分为空白对照组(n=10),模型(100 mg·kg-1·d-1)组(n=10),低(35mg·kg-1·d-1)、中(70mg·kg-1·d-1)和高(140mg·kg-1·d-1)剂量SCL组(n=10)。小鼠连续皮下注射D-半乳糖10周制备D-半乳糖小鼠衰老模型,给予SCL(35、70和140mg·kg-1·d-1)处理10周。水迷宫试验检测小鼠学习记忆能力,Western blotting法检测各组小鼠脑组织中线粒体Bax和Bcl-2蛋白表达、泛素化蛋白(Ub)及微管相关蛋白轻链3(LC3)表达水平的变化;免疫组织化学法观察小鼠大脑皮质和海马组织中LC3蛋白表达。结果:学习记忆能力测试中,与空白对照组比较,模型组小鼠游泳时间延长(P<0.05),错误次数增多(P<0.05);与模型组比较,SCL低、中和高剂量组小鼠游泳时间缩短(P<0.05),错误次数减少(P<0.05);Western blotting法检测,与空白对照组比较,模型组小鼠脑组织中Bax蛋白表达水平增加、Bcl-2蛋白表达水平降低,泛素化蛋白Ub、LC3-Ⅱ/LC3-Ⅰ蛋白表达水平均增加(P<0.05);与模型组比较,SCL低、中和高剂量组小鼠脑组织中Bax蛋白表达水平降低,Bcl-2蛋白表达水平增加,Ub、LC3-Ⅱ/LC3-Ⅰ蛋白表达水平均降低(P<0.05)。免疫组织化学法检测,空白对照组小鼠大脑皮质和海马组织中神经形态正常,神经元细胞浆内无棕黄色颗粒染色,LC3蛋白表达为阴性;模型组小鼠有神经细胞变性,大脑皮质和海马神经元中LC3蛋白阳性细胞数增加(P<0.05);与模型组比较,SCL低、中和高剂量组小鼠神经细胞变性数量明显减少,大脑皮质和海马神经元中LC3蛋白阳性细胞数减少(P<0.05)。结论:SCL具有缓解D-半乳糖诱导的小鼠脑组织衰老作用,其作用机制与SCL调节自噬作用和抑制细胞凋亡有关联。Objective To copy the mouse aging model with D-galactose,and to investigate the role of Schisandra total lignin （SCL）in the mouse brain tissue aging and its mechanism.Methods 50 mice were radomly divided into control group,model group （100 mg·kg-1 ·d-1）,low dose （35 mg·kg-1 ·d-1）of SCL group （SCL-L）, middle dose （70 mg· kg-1 · d-1 ）of SCL group （SCL-M）and high dose （140 mg· kg-1 · d-1 ）of SCL group （SCL-H）（n=10）.D-galactose （100 mg·kg-1 ·d-1 ）was injected into the mice hypodermically for 10 weeks to induce aging models in all the groups except control group,and 35,70,and 140 mg· kg-1 · d-1 SCL were administered for 10 weeks in SCL groups.The learning and memory abilities were measured by the Water Maze test.The expression levels of Bax,Bcl-2,ubiquitin （Ub）,microtubule-associated protein light chain 3 （LC3）in the brain tissue of the mice in various groups were observed by Western blotting method. The LC3 protein expressions in mouse brain cortex and hippocampus were observed by immunohistochemistry.Results In learning and memory test,compared with control group,the swimming time of the mice in model group was increased （P〈0.05）,and the number of errors was increased （P〈0.05）;compared with model group,the swimming time in SCL-L,SCL-M and SCL-H groups was decreased （P〈0.05）and the number of errors was also decreased （P〈0.05）. Compared with control group,the expression level of Bax was increased （P〈0.05）,the expression level of Bcl-2 was decreased （P〈0.05）,the expression levels of Ub and LC3-Ⅱ/LC3-Ⅰ proteins were increased （P〈0.05）in model group;compared with model group,the expression level of Bax was decreased （P〈0.05）,the expression level of Bcl-2 was incerased （P〈0.05）,and the expression levels of Ub and LC3-Ⅱ/LC3-Ⅰ proteins were decreased （P〈0.05）in SCL-L,SCL-M and SCL-H groups.In control group,the neuronal morphology was normal,and none of brown granules were visible in the cytoplasm o

关 键 词：北五味子总木脂素 衰老 自噬 细胞凋亡 

分 类 号：R965[医药卫生—药理学]