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

• 1:贵州医科大学分子生物学重点实验室
• 2:贵州医科大学地方病与少数民族性疾病教育部重点实验室
• 3:贵州医科大学病理学教研室

摘要(Abstract)：

目的:探讨α7胆碱能受体(α7nAChRs)激动剂(PNU)对SD大鼠星形胶质细胞钙调蛋白(CaM)表达水平的影响。方法:将培养第3~4代的星形胶质细胞分为对照组、PNU处理组(PNU组)、α7nAChRs阻断剂(MLA)联合PNU处理组(联合组);对照组不加PNU和MLA处理,PNU组以1、5及10μmol/L的PNU处理星形胶质细胞6、12、18及24 h,联合组以0.05、0.1及0.15μmol/L浓度的MLA预先处理星形胶质细胞2 h后,选择加入上调CaM蛋白表达水平最高的PNU浓度和最佳时间(PNU组筛选后所得)处理细胞;运用蛋白印迹(Western blotting)方法检测3组星形胶质细胞CaM的蛋白表达水平。结果 :与对照组比较,1、5及10μmol/L PNU刺激星形胶质细胞6、12、18及24 h可使CaM蛋白表达水平升高(P<0.05);加入5μmol/L PNU培养12 h后,上调作用最明显;联合组(0.05、0.1及0.15μmol/L的MLA预先处理星形胶质细胞2 h后,再加入PNU 5μmol/L培养12 h)与PNU组比较,星形胶质细胞裂解液中CaM蛋白的表达受到抑制,差异有统计学意义(P<0.05)。结论:PNU激活星形胶质细胞后可提高CaM的蛋白表达水平,MLA可阻断该表达。
Objective: To Detect the influence of α7 cholinergic receptor( α7 nAChRs) PNU on calmodulin( CaM) expression level in SD rat astrocytes. Methods: The 3-4 generations of astrocytes were divided into PNU treatment group( group P),α7 nAChRs blocking agent( MLA) combined with PNU treatment group( group MP),and normal control group( group C). Group C did not receive PNU or MLA. PNU was used to treat astrocytes at doses of 1,5 and 10 μmol/L for 6 h,12 h,18 h and 24 h in group P. In group MP,PNU of optimal concentration was used to cultivate the astrocytes after MLA treated astrocytes at doses of 0. 05,0. 1 and 0. 15 μmol/L for 2 h and in optimal time for cultivation. CaM expression level was detected by western blotting. Results: Ca M expression levels were significantly higher in group P than in group C( P < 0. 05),and the expression peak obtained in12 h after 5 μmol/L of PNU was added. While In group MP,the CaM expression level was markedly lower than that in group P when cells treated by 5 μmol/L of PNU for 12 h( P < 0. 05). Conclusion:The activation of PNU to Astrocytes might increase expression levels of CaM protein,which can be blocked by α7 nAChRs blocker MLA.

关键词(KeyWords)： 星形胶质细胞;α7胆碱能受体;激动剂;钙调蛋白;阿尔茨海默病
astrocytes;α7 cholinergic receptor;agonist;calmodulin;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): 何婧,任真奎,官志忠,禹文峰,吴昌学
HE Jing,REN Zhenkui,GUAN Zhizhong,YU Wenfeng,WU Changxue

DOI: 10.19367/j.cnki.1000-2707.2017.01.002

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