蜂毒明肽对Aβ_(25-35)诱导的阿尔茨海默病模型的体内外作用研究  

The effects of apamin on Aβ_(25-35)-induced Alzheimer's disease model in vivo and in vitro

作　　者：林茂[1,2] 田炳欣 王敏[2,3] 吕延成 胡云[5] 王春梅[1,2] Lin Mao Tian Bingxin Wang Min Lyu Yancheng Hu Yuns Wang Chunmei(Department of Physiology, Zhuhai Campus of Zunyi Medical University, Zhuhai Guangdong 519051, China Zhuhai Key Laboratory of Fundamental and Applied Research in Traditional Chinese Medicine, Zhuhai Guangdong 519041, China Department of Pharmacy,Zhuhai Campus of Zunyi Medical University,Zhuhai Guangdong 519041, China Department of Biochemistry and Molecular Biology, Zhuhai Campus of Zunyi Medical University, Zhuhai Guangdong 519041, China Department of Chemistry, Zhuhai Campus of Zunyi Medical University, Zhuhai Guangdong 519041, China)

机构地区：[1]遵义医学院珠海校区生理学教研室,广东珠海519041 [2]珠海市重点实验室暨中药基础及应用研究实验室,广东珠海519041 [3]遵义医学院珠海校区药学教研室,广东珠海519041 [4]遵义医学院珠海校区生物化学与分子生物学教研室,广东珠海519041 [5]遵义医学院珠海校区化学教研室,广东珠海519041

出　　处：《遵义医学院学报》2017年第4期364-368,373,共6页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:81560562);贵州省教育厅自然科学研究项目(NO:黔教科[2009]0108);珠海市中药基础及应用研究重点实验室开放课题(NO:ZYKL-2016K6)

摘　　要：目的初步探讨蜂毒明肽对Aβ_(25-35)诱导的阿尔茨海默病(AD)模型的体内外干预作用。方法侧脑室注射Aβ_(25-35)构建AD小鼠模型。连续5 d腹腔注射0.2 mg/kg蜂毒明肽。小鼠跳台测试认知功能,免疫组化检测海马CA1区Aβ_(1-42)表达。10μmol/L Aβ_(25-35)诱导海马神经元构建细胞损伤模型。100 nmol/L蜂毒明肽干预24 h后,MTT法检测细胞存活率,Hoechst33342染色检测细胞凋亡率,全细胞膜片钳技术检测海马神经元小电导钙激活性钾通道介导的中间后超极化电流(I_(mAHP))。结果动物实验显示,与模型组比较,蜂毒明肽组小鼠学习成绩错误次数减少,潜伏期延长、记忆成绩错误次数减少,Aβ_(1-42)表达减低(P<0.05)。细胞实验显示,蜂毒明肽组较模型组海马神经元存活率增加,凋亡率下降,I_(mAHP)减小(P<0.05)。结论蜂毒明肽对Aβ_(25-35)诱导的AD模型具有保护作用,其机制可能与减少Aβ表达,降低I_(mAHP)有关。Objective To study the effects of apamin on Aβ25-35-induced Alzheimer＇s disease（ AD） model in vivo and in vitro. Methods AD mice model were established by lateral cerebral ventricle injection of Aβ25-35. Intraperitoneal injection of apamin（ 0. 2 mg/kg） lasted for 5 days. Step down test was used to investigate the effects of apamin on cognitive function,and then immunohistochemistry staining was used to observe the expression of Aβ1-42 in hippocampus CA1 region. The hippocampal neurons injury model was established by Aβ25-35（ 10 μmol/L）. The hippocampal neurons were treated with apamin（ 100 nmol/L） for 24 h. MTT staining was used to assess the viability of cells. Hoechst33342 dye was used to assess the apoptosis of cells. Whole-cell patch-clamp techniques were used to observe the variation of medium-after hyperpolarization current（ ImAHP）mediated by SK of hippocampal neurons. Results In vivo,compared the apamin group with the AD model group,the number of mistakes of learning and memory was decreased,the latent period was longer,and the expression of Aβ1-42 was decreased in hippocampus CA1 region（ P〈0. 05）. In vitro,compared the apamin group with the model group,the viability of hippocampal neurons was increased,the apoptosis was reduced,and the amplitude of ImAHPwas decreased（ P〈0. 05）. Conclusion Apamin has a protective effect on Aβ25-35-induced AD model in vivo and in vitro,which is associated with the reduction of Aβ expression and ImAHP.

关 键 词：蜂毒明肽 Β淀粉样蛋白 阿尔茨海默病 海马神经元 小鼠 

分 类 号：R741[医药卫生—神经病学与精神病学] R285.5[医药卫生—临床医学]