任真奎,何婧,吴昌学,官志忠,禹文峰
REN Zhenkui,HE Jing,WU Changxue,GUAN Zhizhong,YU Wenfeng

• 1:贵州医科大学分子生物学重点实验室

摘要(Abstract)：

目的:探讨尼古丁激活SD大鼠星形胶质细胞α7胆碱能受体(α7 n AChRs)后内源性B-晶状体蛋白(Cryab)表达。方法:分离24 h新生乳鼠大脑皮质,原代培养并鉴定为星形胶质细胞后,将培养第3~4代的星形胶质细胞分为正常对照组、尼古丁处理组和α7 n AChRs阻断剂(MLA)联合尼古丁处理组;正常对照组不加尼古丁和MLA,尼古丁处理组以1、5及10μmol/L浓度的尼古丁处理星形胶质细胞6、12、18及24 h,MLA联合尼古丁处理组以0.05、0.1、0.15及0.2μmol/L浓度的MLA预先处理星形胶质细胞2 h后,再加入尼古丁处理组最佳浓度的尼古丁和最佳培养时间培养细胞;运用蛋白印迹(Western blotting)方法检测3组星形胶质细胞内源性Cryab蛋白的表达。结果:与正常对照组比较,尼古丁处理组的星形胶质细胞内源性Cryab蛋白上调(P<0.05);与尼古丁处理组比较,MLA联合尼古丁处理组的星形胶质细胞内源性Cryab蛋白上调受到明显抑制(P<0.01)。结论:尼古丁激活星形胶质细胞α7n AChRs可上调内源性Cryab蛋白表达,该表达可被MLA阻断。
Objective: To investigate the expression of endogenous Cryab protein after the activation of α7 n AChRs by nicotine in SD rat astrocytes. Methods: The rat cerebral cortex of 24 h neonatal was isolated. After primary culture and identification of astrocytes,the astrocytes cultivated for 3 ~ 4 generation were divided into normal control group,nicotine treatment group and α7 n AChRs blocking agent( MLA) plus nicotine treatment group. Normal control group did not receive nicotine and MLA,1,5and 10 μmol / L nicotine was used to treat astrocytes for 6,12,18 and 24 h in nicotine treatment group,and in MLA plus nicotine treatment group 0. 05,0. 1,0. 15 and 0. 2 μmol / L MLA treated astrocytes for 2 h previously and nicotine of optimal concentration was used to cultivate the astrocytes in optimal cultivation time. Western blotting was used to detect the expression of endogenous Cryab protein in astrocytes the three groups. Results: Compared with normal control group,the endogenous Cryab protein expression up-regulated significantly in nicotine treatment group( P < 0. 05). Compared with nicotine treatment group,the up-regulation of endogenous Cryab protein expression in MLA plus nicotine treatment group was significantly inhibited( P < 0. 01). Conclusions: The activation of α7n AChRs of astrocytes by nicotine can up-regulate endogenous Cryab protein expression,which can beblocked by α7 n AChRs blocker MLA.

关键词(KeyWords)： 星形胶质细胞;B-晶状体蛋白;α7胆碱能受体;尼古丁;阿尔茨海默氏病
astrocytes;Cryab;α7 cholinergic receptor;nicotine;Alzheimer's disease

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81360199);; 教育部科学技术研究项目(213032A);; 贵州省国际科技合作项目[黔科合外G字(2013)7026号]

作者(Author): 任真奎,何婧,吴昌学,官志忠,禹文峰
REN Zhenkui,HE Jing,WU Changxue,GUAN Zhizhong,YU Wenfeng

DOI: 10.19367/j.cnki.1000-2707.2016.08.002

参考文献(References)：

• [1]Zhu Z,Li R,Stricker R,et al.Extracellularα-crystallin protects astrocytes from cell death through activation of MAPK,PI3K/Akt signaling pathway and blockade of ROS release from mitochondria[J].Brain Research,2015(1620):17-28.
• [2]Ousman SS,Tomooka BH,Noort JMV,et al.Protective and therapeutic role forαB-crystallin in autoimmune demyelination[J].Clinical Immunology,2007(Suppl):S139.
• [3]Wilhelmus MMM,Waal RMW,Verbeek MM.Heat shock proteins and amateur chaperones in amyloid-beta accumulation and clearance in Alzheimer’s Disease[J].Molecular Neurobiology,2007(3):203-16.
• [4]Toledo JB,Bjerke M,Da X,et al.Nonlinear association between cerebrospinal fluid and florbetapir f-18β-amyloid measures across the spectrum of Alzheimer Disease[J].Jama Neurology,2015(5):78-85.
• [5]Ittner LM,Ke YD,Delerue F,et al.Dendritic function of tau mediates amyloid-βToxicity in Alzheimer's Disease mouse models[J].Cell,2010(3):387-397.
• [6]Dehle FC,Heath E,Musgrave IF,et al.αB-Crystallin inhibits the cell toxicity associated with amyloid fibril formation byκ-casein and the amyloid-βpeptide[J].Cell Stress&Chaperones,2010(6):1013-1026.
• [7]Shammas SL,Waudby CA,Wang S,et al.Binding of the molecular chaperoneαB-crystallin to Aβamyloid fibrils inhibits fibril elongation[J].Biophysical Journal,2011(7):1681-1689.
• [8]Wilhelmus MM,Boelens WC,Otte-Hller I,et al.Small heat shock proteins inhibit amyloid-βprotein aggregation and cerebrovascular amyloid-βprotein toxicity[J].Brain Research,2006(4):67-78.
• [9]Fu W,Shi D,Westaway D,et al.Bioenergetic mechanisms in astrocytes may contribute to amyloid plaque deposition and toxicity[J].Journal of Biological Chemistry,2015(20):98-105.
• [10]Heneka MT,M Kerry O,Dick T,et al.Inflammatory processes in Alzheimer's disease[J].Journal of Neuroimmunology,2007(1-2):69-91.
• [11]Sofroniew MV,Vinters HV.Astrocytes:biology and pathology[J].Acta Neuropathologica,2010(1):7-35.
• [12]张敬军.星形胶质细胞的研究[J].中国药理学通报,2006(7):788-791.
• [13]Béduer A,Joris P,Mosser S,et al.Detection of Alzheimer's disease amyloid-beta plaque deposition by deep brain impedance profiling[J].Journal of Neural Engineering,2015(2):78-85.
• [14]Rodríguez JJ,Olabarria M,Chvatal A,et al.Astroglia in dementia and Alzheimer's disease[J].Cell Death&Differentiation,2009(8):378-385.
• [15]Russo P,Del Bufalo A,Frustaci A,et al.Beyond acetylcholinesterase inhibitors for treating Alzheimer's disease:α7-n AChR agonists in human clinical trials[J].Current Pharmaorutioal Design,2014(38):6014-6021.
• [16]Corinne B,Tristan R,Samuel D.Banister,et al.Structure-activity relationship studies of SEN12333 analogues:Determination of the optimal requirements for binding affinities atα7 n AChRs through incorporation of known structural motifs[J].European Journal of Medicinal Chemistry,2015(95):277-301.

文章评论(Comment)：