洛伐他汀上调体外培养神经细胞毒蕈碱型乙酰胆碱受体及对抗β-淀粉样肽对该受体表达的抑制作用

Lovastatin may up-regulate expression of muscarinic acetylcholine receptors and against the neurotoxicity of β-amyloid peptide on the receptors

目的观察洛伐他汀是否对毒蕈碱型乙酰胆碱受体(mAChRs)有上调作用,探讨其是否具有抗β-淀粉样肽(Aβ)的作用。方法选择原代培养的大鼠海马神经细胞和SH-SY5Y细胞,采用洛伐他汀和Aβ寡聚体(AβOs)分别或联合处理培养细胞24~48 h后,采用蛋白印迹法和实时荧光定量PCR(Real-time PCR)检测细胞中M1 mAChR和M3 mAChR蛋白及mRNA表达水平。结果免疫荧光染色显示原代大鼠海马神经细胞的体外培养纯度达85%以上;不同浓度洛伐他汀处理神经细胞24 h后,M1 mAChR和M3 mAChR蛋白及mRNA表达水平均明显高于对照组(P<0.05);0.5μmol/L AβOs处理神经细胞48 h后,其M1 mAChR和M3 mAChR蛋白及mRNA表达水平均明显下降(P<0.05);但预先用0.1μmol/L洛伐他汀处理神经细胞24 h,可减轻AβOs导致的M1 mAChR和M3 mAChR表达水平下降。结论洛伐他汀可能具有上调mAChRs表达水平的作用,对抗Aβ对该受体的作用。

Objective To investigate the influence of lovastatin on muscarinic acetylcholine receptors （mAChRs）,and reveal whether the drug attenuate the inhibited effect of-amyloid peptide （Aβ） on the receptors.Methods The primary cultured rat hippocampal neurons and SH-SY5Y cells were exposed to lovastatin and Ab oligomers （AβOs） with alone or both treatments for 24~48 h.The protein levels of M1 and M3 mAChR were examined by Western blot,and the mRNA levels were tested by Real-time PCR.Results The purity of primary cultured rat hippocampal neurons was over 85%;by exposures of different concentrations of lovastatin for 24 h,the protein and mRNA levels of M1 and M3 mAChR were higher than those of control group,respectively （P〈0.05）;as compared with those of control group,the protein and mRNA levels of M1 and M3 mAChR were obviously decreased in the treatment with 0.5 μmol/L AβOs for 48 h （P〈0.05）;while the cells were pretreated with 0.1 μmol/L lovastatin for 24 h before the treatments by AβOs,the decreased protein and mRNA levels of M1 and M3 mAChR were all alleviated.Conclusions Lovastatin might up-regulate expression of mAChRs and attenuate the inhibited effect of Aβ on the receptors.