摘要
目的研究六味地黄丸含药血清(MSLDP)对β-淀粉样蛋白(Aβ)25-35损伤大鼠肾上腺嗜铬细胞瘤细胞(PC12细胞)的保护作用,探讨六味地黄丸在防治阿尔茨海默病(AD)中的作用机制。方法 2012年3月—2013年4月选取SPF级雄性SD大鼠20只,按照随机数字表法将20只雄性SD大鼠分为对照组、给药组,每组10只。制备MSLDP及Aβ25-35母液备用。根据Aβ25-35、MSLDP不同干预浓度将处于对数生长期PC12细胞分为0、10、20、40 μmol/LAβ25-35组和0%、2.5%、5.0%、10.0%、20.0%、30.0%MSLDP组,另取处于对数生长期PC12细胞分为正常对照组(不加Aβ25-35和MSLDP)、模型组(加入20 μmol/L Aβ25-35制备AD模型)、MSLDP干预组(加入20%MSLDP+20 μmol/L Aβ25-35),显微镜下观察0%、2.5%、5.0%、10.0%、20.0%、30.0%MSLDP组及正常对照组、模型组、MSLDP干预组PC12细胞形态。采用MTT法检测PC12细胞存活率,同上分组方法及步骤采用Western blotting法检测PC12细胞血管内皮生长因子B(VEGF-B)蛋白表达水平。结果 0 μmol/L Aβ25-35组PC12细胞存活率高于10、20、40 μmol/L Aβ25-35组(P<0.05);10 μmol/L Aβ25-35组PC12细胞存活率高于20、40 μmol/L Aβ25-35组(P<0.05);20 μmol/L Aβ25-35组PC12细胞存活率高于40 μmol/L Aβ25-35组(P<0.05)。0、10 μmol/L Aβ25-35组VEGF-B蛋白表达水平高于20、40 μmol/L Aβ25-35组(P<0.05);20 μmol/L Aβ25-35组VEGF-B蛋白表达水平高于40 μmol/L Aβ25-35组(P<0.05)。0%MSLDP组PC12细胞存活率低于2.5%、5.0%、10.0%、20.0%、30.0%MSLDP组(P<0.05);2.5%MSLDP组PC12细胞存活率低于5.0%、10.0%、20.0%、30.0%MSLDP组(P<0.05);5.0%MSLDP组PC12细胞存活率低于10.0%、20.0%、30.0%MSLDP组(P<0.05);10.0%MSLDP组PC12细胞存活率低于20.0%、30.0%MSLDP组(P<0.05)。20.0%MSLDP组PC12细胞存活率低于30.0%MSLDP组(P<0.05)。显微镜下示0%MSLDP组PC12细胞形态规则,大小均一;2.5%、5.0%MSLDP组PC12细胞数目增多,形态正常;10.0%MSLDP组PC12细胞聚集成片状生长;20.0%、30.0%MSLDP�
Objective To explore the effects and mechanisms of Liuwei dihuang pill （LDP） in the prevention and treatment of alzheimer disease （AD） by studying the protective effects of medicated serum of Liuwei Dihuang Pill （MSLDP） on rat adrenal pheochromocytoma cells （PC12） injured by amyloid - beta peptide 25 - 35 （ Aβ25 - 35 ） . Methods This study was conducted from March 2012 to April 2013. Twenty SPF grade SD male rats were selected and divided into the control group and the drug group according to the random number table, 10 for each group. MSLDP and Aβ25 - 35 were prepared. According to the intervention concentration of Aβ25 -35 and MSLDP, PC12 ceils in logarithmic growth phase were taken and divided into 0, 10, 20, 40μmol/L Aβ25 -35 group and 0%, 2. 5%, 5.0%, 10.0%, 20. 0%, 30. 0% MSLDP group. Then we took other PC12 cells in logarithmic growth phaseand divided into the normal control group （ without the intervention of Aβ25 - 35 and MSLDP） , model group （ receiving intervention of 20 μmol/L Aβ25 - 35 for the preparation of AD model） , and MSLDP intervention group （receiving intervention of 20% MSLDP ＋ 20 μmol/L Aβ25 -35 ） . The survival rate of PC12 cells in all the groups was detected by MTT. Then we took other PC12 cells in logarithmic growth phase and obtained the same groups by using the above grouping methods and steps. The morphology of PC12 ceils in all MSLDP groups, normal control group, model group and MSLDP intervention group were observed under the microscope. The express ion level of vascular endothelial growth factor B （ VEGF - B） in PC12 cells in these groups was detected by western blotting. Results The survival rate of PC12 cells in 0μmol/ L Aβ25 -35 group was higher than that in the other three Aβ25 -35 treated groups （P 〈0. 05） ; the survival rate of PC12 cells in 10μmol/L Aβ25 -35 group was higher than that of 20, 40 μmol/L AI325 -35 group （P 〈 0. 05 ） ; the survival rate of PC12 cells in 20μmol/L Aβ25 -35 group was higher than t
作者
宋军营
袁永
王丛笑
张钟允
宫洪涛
张振强
刘文弟
高爱社
SONG Jun - ying YUAN Yong WANG Cong - xiao ZHANG Zhong - yun GONG Hong - tao ZHANG Zhen - qiang LIU Wen - di GAO Ai - she(Immunology Laboratory of Chinese Medicine, Henan University of Chinese Medicine, Zhengzhou 450046, China The Third Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450008, China The First Affiliated Hospital of Henan University of TCM, Zhengzhou 450000, China School of Basic Medical Sciences, Henan University of Chinese Medicine, Zhengzhou 450046, China)
出处
《中国全科医学》
CAS
北大核心
2017年第30期3763-3769,共7页
Chinese General Practice
基金
国家自然科学基金联合基金项目(U1504829)
河南省基础与前沿科学研究项目(14230041023
162300410127)
河南省高校科技创新人才支持计划(13HASTIT027)
河南省科技攻关项目(16102310182)
关键词
阿尔茨海默病
六味地黄汤
血管内皮生长因子类
淀粉样β肽类
Alzheimer disease
Liuwei dihuang decoction
Vascular endothelial growth factors
Amyloid beta - peptides
