固本健脑液通过调节APP代谢途径对AD大鼠发挥神经保护作用  

Study on Neuroprotective Effect of Guben Jiannao Liquid on AD Rats by Regulating APP Metabolism Pathway

作　　者：曾楚华 刘思文 王雨 谢喆尧 张婧凡 蔡元钦 王曦 ZENG Chu-hua;LIU Si-wen;WANG Yu;XIE Zhe-yao;ZHANG Jing-fan;CAI Yuan-qin;WANG Xi(School of Basic Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China;Health Science Center,Hubei Minzu University,Enshi 445000,China;Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases of Hubei Minzu University,Enshi 445000,China;College of Pharmacy,Hubei University of Medicine,Shiyan 442000,China)

机构地区：[1]云南中医药大学基础医学院,云南昆明650500 [2]湖北民族大学医学部,湖北恩施445000 [3]湖北民族大学风湿性疾病发生与干预湖北省重点实验室,湖北恩施445000 [4]湖北医药学院药学院,湖北十堰442000

出　　处：《中药材》2023年第12期3097-3103,共7页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金地区基金项目(82060831)。

摘　　要：目的:基于APP代谢途径探讨固本健脑液对AD大鼠的神经保护作用。方法:SPF级雄性SD大鼠50只,随机分为正常对照组、模型组、多奈哌齐阳性对照组及固本健脑液低、高剂量组,每组10只。除正常对照组外各组大鼠均予双侧海马CA1区注射Aβ_(1-42)建立AD模型。造模后第4天开始灌胃相应药物,正常对照组与模型组灌胃相应体积生理盐水。4 w后行Morris水迷宫行为学实验;行为学实验结束后取材,HE染色、尼氏染色、TUNEL法检测大鼠海马CA3、CA1区神经细胞形态、数量等;ELISA检测大鼠海马中Aβ_(1-42)含量,Western Blot检测大鼠海马中APP、ADAM10、BACE1、PS1、Bax、Caspase-3蛋白表达。结果:与正常对照组比较,模型组大鼠Morris水迷宫实验逃避潜伏期时间显著延长,且航行轨迹杂乱无序,穿过原有平台次数及第三象限停留时间显著缩短(P<0.01),海马CA3、CA1区细胞核深染,细胞数量少,排列稀疏紊乱,形态不规则;尼氏染色较淡,细胞排列稀疏散乱,细胞核破裂不完整,尼氏小体数量明显减少,凋亡细胞明显增多,海马中Aβ_(1-42)水平及BACE1、PS1、Bax、Caspase-3蛋白表达显著升高,ADAM10蛋白表达显著降低(P<0.01)。与模型组比较,多奈哌齐组及固本健脑液高剂量组大鼠逃避潜伏期显著缩短,海马中Aβ_(1-42)含量显著降低,穿越平台次数和第三象限停留时间显著延长(P<0.05或P<0.01),各给药组大鼠海马CA3、CA1区组织病理学改善明显,尼氏小体数量明显增多,凋亡细胞明显减少;海马中BACE1、PS1、Caspase-3蛋白表达显著降低,固本健脑液高剂量组海马Bax蛋白表达显著降低,固本健脑液各剂量组海马ADAM10蛋白表达显著升高(P<0.05或P<0.01)。结论:固本健脑液可改善AD大鼠的学习记忆能力,其机制可能是通过调节APP代谢途径上相关因子的表达,从而起到保护大鼠海马神经细胞的作用。Objective:To explore the neuroprotective effect of Guben jiannao liquid on AD model rats based on APP metabolic pathway.Methods:Fifty SPF grade male SD rats were randomly divided into normal control group,model group,donepezil positive control group and low,high doses of Guben jiannao liquid groups,with 10 rats in each group.In addition to the normal control group,all rats in other group were injected with Aβ_(1-42) in hippocampal CA1 region to establish AD model.On the 4th day after the modeling,the corresponding drugs were given by intragastric administration,the rats in normal control group and model group were given the corresponding volume of normal saline.Four weeks later,Morris water maze behavior test was performed.The morphology and quantity of nerve cells in CA3 and CA1 regions of hippocampus were detected by HE staining,Nissl staining and TUNEL method after behavioral study.The contents of Aβ_(1-42) in hippocampus were detected by ELISA,and the expressions of APP,ADAM10,BACE1,PS1,Bax and Caspase-3 proteins in the hippocampus were detected by Western Blot.Results:Compared with the normal control group,the escape latency time of the rats in model group was significantly extended,the sailing track was clear and disordered,the number of crossing the original platform and the time of staying in the third quadrant were significantly shortened(P<0.01),the nuclei of CA3 and CA1 regions of hippocampus were deeply stained,the number of cells was small,the arrangement was sparse and disordered,and the shape was irregular.The Nissl staining was light,the cell arrangement was sparse and scattered,and the nucleus was broken incomplete.The number of Nissl bodies was significantly decreased,the apoptosis of apoptotic cells was significantly increased,the level of Aβ_(1-42) and the expressions of BACE1,PS1,Bax and Caspase-3 in hippocampus were significantly increased,and the protein expression of ADAM10 was significantly decreased(P<0.01).Compared with model group,the escape latency of rats in Donepezil group and Gu

关 键 词：阿尔兹海默病 固本健脑液 APP代谢 神经保护 

分 类 号：R285.5[医药卫生—中药学]