谭龙春,赵亮,邓成敏,刘仙红,官志忠
TAN Longchun,ZHAO Liang,DENG Chengmin,LIU Xianhong,GUAN Zhizhong

• 1:贵州医科大学贵州省医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:贵州医科大学微生物学教研室
• 4:贵州医科大学附院病理科

摘要(Abstract)：

目的:观察洛伐他汀对大鼠原代海马神经元毒蕈碱样型乙酰胆碱受体(mAChRs)及细胞外信号调节激酶(ERK)信号通路的作用。方法:选择大鼠原代培养海马神经细胞,(1)将细胞分为对照组、Atropine处理组及U0126处理组,观察mAChRs拮抗剂Atropine及ERK1/2通路抑制剂U0126对M1 mAChR和ERK1/2信号通路的影响;(2)将细胞分为对照组、Atropine及洛伐他汀处理组、洛伐他汀组、U0126及洛伐他汀处理组,观察洛伐他汀联合Atropine或U0126对M1 mAChR和ERK1/2信号通路的影响;采用蛋白印迹法(Western blotting)检测M1 mAChR、总ERK1/2(p-ERK1/2)及磷酸化ERK(phospho-ERK1/2)蛋白表达水平。结果:(1)与对照组相比,Atropine和U0126分别处理细胞后,phospho-ERK1/2蛋白表达水平明显降低(P<0.05),M1 mAChR和pERK1/2表达水平未见明显改变;(2)与对照组相比,0.1μmol/L洛伐他汀处理神经细胞,M1 mAChR及phosphoERK1/2蛋白表达水平明显增高(P<0.05);与洛伐他汀处理组相比,联合应用洛伐他汀与Atropine处理细胞,M1 mAChR及phospho-ERK1/2蛋白表达水平的升高未受到明显影响,但联合应用洛伐他汀与U0126处理细胞,phospho-ERK1/2蛋白表达水平明显下降(P<0.01),M1 mAChR的升高未受到明显影响。结论 :洛伐他汀能够活化ERK1/2信号通路,并上调mAChRs表达水平,但洛伐他汀上调mAChRs表达水平的作用与ERK1/2信号通路无关。
Objective: To investigate the influence of lovastatin on muscarinic acetylcholine receptors(m AChRs) and extracellular signal-regulated kinase( ERK) signaling pathway. Methods: The primary cultured rat hippocampal neurons were used in 2 ways.( 1) Cells were treated with antagonist of m AChRs atropine or inhibitor of ERK U0126 for 2 h,and then divided into three groups according to the following treatment: control group,atropine group( 100 μmol/L atropine),and U0126 group(10 μmol/L U0126);( 2) After treated by atropine or U0126 alone or combined with lovastatin,Cells were divided into four groups: control group,atropine plus lovastatin group,and U0126 plus lovastatin group. The protein expression of M1 mAChR,p-ERK1/2,and phospho-ERK1/2 were detected by Western blotting. Results: Compared with control group,100 μmol/L atropine or 10 μmol/L U0126 reduced the level of phospho-ERK1/2( P < 0. 05),but did not influence the level of M1 mAChR and p-ERK1/2. Compared with control group,0. 1 μmol/L lovastatin increased the expression levels of M1 mAChR and phospho-ERK1/2( P < 0. 05),and when the cells were treated with lovastatin and atropine,the elevated expression of M1 m AChR and phospho-ERK1/2 did not changed. But the expression of phospho-ERK1/2 resulted from lovastatin markedly decreased( P < 0. 01) when the cells were treated with lovastatin and U0126. Conclusion: Lovastatin might activate ERK signaling path way,and up-regulate expression of M1 mAChR. But there is no relationship between these 2effects.

关键词(KeyWords)： 洛伐他汀;毒蕈碱型乙酰胆碱受体;ERK1/2信号通路;细胞培养;海马;神经细胞;大鼠,Sprague-Dawley
lovastatin;muscarinic acetylcholine receptors;ERK1/2 signaling pathway;cell culture;hippocampe;neurons;rats,Sprague-Dawley

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81260173);; 教育部“长江学者和创新团队发展计划资助”(IRT13058);; 贵州省科技计划[黔科合重大专项字(2014)6008号];; 贵州省创新计划项目[黔教合协同创新中心(2014)06]

作者(Author): 谭龙春,赵亮,邓成敏,刘仙红,官志忠
TAN Longchun,ZHAO Liang,DENG Chengmin,LIU Xianhong,GUAN Zhizhong

DOI: 10.19367/j.cnki.1000-2707.2017.01.001

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