灯盏细辛对神经细胞尼古丁受体表达的影响及其神经保护作用研究_药学论文.docVIP

灯盏细辛对神经细胞尼古丁受体表达的影响及其神经保护作用研究_药学论文 灯盏细辛对神经细胞尼古丁受体表达的影响及其神经保护作用研究_药学论文 作者：齐晓岚，黄勇，王永林，王爱民，官志忠 【摘要】 　　目的研究中药灯盏细辛对神经母细胞瘤 （SH-SY5Y） 细胞神经型尼古丁受体表达的影响及其对抗β淀粉样蛋白 （Aβ）神经毒性作用机制。方法采用比色法测定细胞 MTT 还原率筛选药物浓度安全范围，选取一定浓度的灯盏细辛乙醇提取物醋酸乙酯部位处理 SH-SY5Y 细胞后，再加入 Aβ1-42 处理，测定 MTT 还原率，蛋白质印迹法（Western blotting ）方法测定神经型尼古丁受体 α3、α7 蛋白表达的变化；采用改进的Ellman比色法测定细胞培养基中乙酰胆碱酯酶（Acetylcholinesterase，AChE）及丁酰胆碱酯酶（Butyrylcholinesterase，BuChE）活性。结果灯盏细辛乙醇提取物醋酸乙酯部位能提高细胞 α3 及 α7 尼古丁受体亚单位蛋白质表达水平；降低细胞乙酰胆碱酯酶活性；并能对抗 Aβ 引起的MTT还原率降低。结论灯盏细辛乙醇提取物醋酸乙酯部位能上调尼古丁受体亚单位蛋白表达水平，降低细胞乙酰胆碱酯酶活性，且能对抗 Aβ 的神经毒性作用，在阿尔茨海默氏病的治疗中可能起到一定作用。 【关键词】 灯盏细辛； SH-SY5Y细胞； 尼古丁受体； β-淀粉样蛋白； 胆碱酯酶； 阿尔茨海默氏病 　　Abstract：ObjectiveTo investigate the effects of Herba Erigeromtis on the expression of nicotinic acetylcholine receptor (nAChR) and the anti-neurotoxicity mechanism of the drug by using neuroblastoma SH-SY5Y cells treated with beta-amyloid peptide.MethodsThe SH-SY5Y cells were treated by Herba Erigeromitis, and then exposed to Aβ1-42. MTT reduction assay was used to choos a safe-dose concentration of Herba Erigeromitis and to understand the influences of the drugs on cellular viability resulted from the treatment of Aβ; expressions of α3 and α7 nAChR subunits at protein level were detected by Western-blotting. The levels of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activities in cell culture mediums were meatured by an improved Ellman‘s colorimetric method. ResultsHerba Erigeromtis stimulated increases of α7 and α3 nAChR proteins in the cultured cells; decreased the activity of AChE and inhibited the decreased MTT reduction resulted from Aβ. ConclusionHerba Erigeromtis can upregulate the expression of nAChR subunits at protein level, decrease the activity of AChE and prevent the decreased MTT reduction induced by Aβ. The drugs might play an important role in therapeutic strategy of AD. 　　Key words： SH-SY5Y cells; nAChR; β-amyloid pepitide; Alzheimer disease; cholinesterase; Herba Erigeromtis 　　阿尔茨海默氏病（Alzheimers disease， 简称AD）是一种主