摘要
目的:探究二精丸对去卵巢+D-半乳糖联合β-淀粉样蛋白1-40(Aβ1-40)所致肾阴虚阿尔茨海默病(AD)大鼠生物学基础的影响。方法:大鼠去卵巢后随机分为5组,即模型组、多奈哌齐组、二精丸高、中、低剂量组,每组11只,另设11只为假手术组,术后1周,连续腹腔注射D-半乳糖7周,术后4周,单侧海马注射Aβ1-40。术后3周开始灌胃给药,模型组与假手术组灌胃生理盐水,二精丸高、中、低剂量组分别用对应浓度灌胃给药(9.0,4.5,2.25 g·kg^-1),多奈哌齐组灌胃1.0 mg·kg^-1多奈哌齐,给药体积为10 mL·kg^-1,每日1次,连续给药35 d。给药第31天,Y迷宫检测大鼠学习能力。末次给药后麻醉大鼠,分离大鼠海马,尼氏染色观察形态学。提取海马蛋白,Nanol-ESI液相-质谱联用系统检测蛋白,Protein Discovery软件鉴定蛋白,SIEVE软件对海马蛋白进行相对定量定性分析,PANTHER Classification System数据库对差异蛋白进行基因本体(GO)分析,String分析和京都基因与基因组百科全书(KEGG)富集信号通路。结果:与模型组大鼠比较,二精丸高、中剂量组大鼠Y迷宫正确交替率明显提高(P<0.05),海马CA1区神经元数量明显提高(P<0.01);二精丸高剂量组的差异蛋白(Ratio>1.5或Ratio<0.5)有胰岛素样生长因子-1受体(IGF-1R)等115个,中剂量组差异蛋白有突触素等94个,多奈哌齐组差异蛋白有乙酰辅酶A乙酰转移酶等87个。GO分析发现这些蛋白主要分为微管相关蛋白、热休克蛋白、能量代谢相关蛋白等与AD相关蛋白;KEGG分析发现上述差异蛋白共涉及Dopaminergic synaps等93条信号通路。结论:二精丸可能是通过磷脂酰肌醇-3激酶/蛋白激酶B(PI3K/Akt)信号通路、胰岛素信号通路、酸活化蛋白激酶(AMPK)信号通路、雌激素信号通路,Dopaminergic synapse等多条通路相关蛋白的表达,从而达到防治肾阴虚AD的目的。
Objective:To explore the effect of Erjingwan on the biological basis of kidney yin deficiency Alzheimer's disease(AD)rats induced by ovariectomy+D-galactose combined with amyloid beta1-40(Aβ1-40).Method:After ovariectomy,rats were randomly divided into five groups:model group,positive group,Erjingwan high,medium and low dose group,11 rats in each group,and 11 rats in sham operation group.One week after operation,D-galactose was injected intraperitoneally for 7 weeks,and four weeks after operation,Aβ1-40 was injected unilaterally into hippocampus.The rats in model group and sham-operation group were given saline by intragastric administration 3 weeks after operation.The rats in high,middle and low dose groups of Erjingwan were given corresponding concentration(9.0,4.5,2.25 g·kg^-1).The rats in positive control group were given Donepezil 1.0 mg·kg^-1 with dosage of 10 mL·kg^-1 once a day for 35 consecutive days.After 30 days of administration,the learning ability of the rats were examined using a Y-maze.The hippocampus tissues of rats were isolated.The morphology of hippocampus was observed by Nissl staining.The proteins were detected by Nanol-ESI liquid-mass spectrometry system and identified by protein Discovery software.Relative quantitative and qualitative analysis of differential proteins in hippocampus was performed by SIEVE software,and Gene Ontology of differential protein was performed by PANTHER Classification System database.String analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were performed on the differential proteins.Result:Compared with model group,the correct rate of Y maze in the high and middle dose groups of Erjingwan was significantly raised(P<0.05),and the number of neurons in the hippocampal CA1 area was significantly increased(P<0.01).115 differential proteins(Ratio>1.5 or Ratio<0.5)such as Insulin-like growth factor 1 receptors(IGF-1R)were found in the high-dose group of the Erjingwan group as well as 94 differential proteins such as Synaptophysin expresse
作者
黄丽萍
杨喜洋
燕波
邱小鹏
严斐霞
孙梦盛
谢永艳
官扬
周茂福
陈耀辉
HUANG Li-ping;YANG Xi-yang;YAN Bo;QIU Xiao-peng;YAN Fei-xia;SUN Meng-sheng;XIE Yong-yan;GUAN Yang;ZHOU Mao-fu;CHEN Yao-hui(Science and Technology College,School of Pharmacy,Jiangxi University of Traditional Chinese Medicine(TCM),Nanchang 330004,China;Jiangxi Provincial People's Hospital,Nanchang 330006,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第9期15-22,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
江西省自然科学基金项目(20171BAB205083)
江西省主要学科学术和技术带头人项目(20182BCB22005)
江西省卫计委中医药科研计划项目(2016A023)
江西省研究生创新专项(YC2017-S356)。
