中国全科医学2017，Vol.20Issue(30)：3763-3769,7.DOI:10.3969/j.issn.1007-9572.2017.00.067
六味地黄丸含药血清对β-淀粉样蛋白损伤大鼠肾上腺嗜铬细胞瘤细胞的保护作用研究
Protective Effects of Medicated Serum of Liuwei Dihuang Pill on Aβ25-35-injured Rat PC12 Cells
摘要
Abstract
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 (A325-35).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 cells in logarithmic growth phase were taken and divided into 0,10,20,40 μmol/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 MTF.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 cells 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 0 μmol/ 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 10 μmol/L Aβ25-35 group was higher than that of 20,40 μmol/L Aβ25-35 group (P ＜0.05);the survival rate of PC12 cells in 20 μnol/L Aβ25-35 group was higher than that of 40 μmol/L Aβ25-35 group (P ＜ 0.05).The protein expression of VEGF-B was higher in 0,10 μmol/L Aβ25-35 group than in the 20,40 μmol/L Aβ25-35 group (P ＜0.05),and it was higher in the 20 μmol/L Aβ25-35 group than in the 40 μmol/L Aβ25-35 group (P ＜ 0.05).The survival rate of PC12 cells was lower in the 0％ MSLDP group than that in other MSLDP groups (P ＜ O.05);the survival rate of PC12 cells in the 2.5％ MSLDP group was lower than that of the 5.0％,10.0％,20.0％,30.0％ MSLDP group (P ＜ 0.05);the survival rate of PC12 cells in the 5.0％ MSLDP group was lower than that of 10.0％,20.0％,30.0％ MSLDP group (P ＜ 0.05);the survival rate of PC12 cells in the 10.0％ MSLDP group was lower than that of the 20.0％,30.0％ MSLDP group (P ＜ O.05);the survival rate of PC12 cells in the 20.0％ MSLDP group was lower than that of the 30.0％ MSLDP group (P ＜ 0.05).Microscope observation showed that,the PC12 cells in the 0％ MSLDP group were regular in shape and uniform in size;the PC12 cells in 2.5％,5.0％ MSLDP group were increased but with normal shape;PC12 cells in 10.0％,20.0％,30.0％ MSLDP group displayed aggregated and 1amellar growth with small protrusions on the edge and differentiated trend.The protein expression of VEGF-B was lower in 0％,2.5％,5.0％ MSLDP group than that in 10.0％,20.0％,30.0％ MSLDP group (P ＜ 0.05),and it was lower in the 10.0％ MSLDP group than in 20.0％ MSLDP group (P ＜ 0.05),but it was higher in the 20.0％ MSLDP group than in the 30.0％ MSLDP group (P ＜0.05).The survival rate of PC12 cells was higher in the normal control group and MSLDP intervention group than that in the model group (P ＜ 0.05).Microscope observation demonstrated that,the PC12 cells in the normal control group were regular in shape and uniform in size;the PC12 cells in the model group were reduced and irregular,presenting cellular atrophy or death as well as cellular debris;the PC12 cells in MSLDP intervention group were significantly increased,and part of them were stretched with protrusions.The protein expression of VEGF-B in normal control group was higher than that in the model group and MSLDP intervention group (P ＜ 0.05);the protein expression of VEGF-B in model group was lower than that of MSLDP intervention group (P ＜ 0.05).Conclusion MSLDP can increase theactivity of the Aβ325-35-injured PC12 cells and protect them,the mechanism might be related to the increase of the protein expression of VEGF-B.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Key words
Alzheimer disease/Liuwei dihuang decoction/Vascular endothelial growth factors/Amyloid beta -peptides分类
医药卫生引用本文复制引用
宋军营,袁永,王丛笑,张钟允,宫洪涛,张振强,刘文弟,高爱社..六味地黄丸含药血清对β-淀粉样蛋白损伤大鼠肾上腺嗜铬细胞瘤细胞的保护作用研究[J].中国全科医学,2017,20(30):3763-3769,7.基金项目
国家自然科学基金联合基金项目(U1504829) （U1504829）
河南省基础与前沿科学研究项目(14230041023,162300410127) （14230041023,162300410127）
河南省高校科技创新人才支持计划(13HASTIT027) （13HASTIT027）
河南省科技攻关项目(16102310182) （16102310182）
