摘要
目的利用ChAT启动子荧光素酶报告系统对中药小分子化合物进行筛选,以期找到能够促进ChAT表达的化合物。方法用构建的ChAT启动子荧光素酶报告体系从400余种中药小分子化合物中筛选ChAT启动子激活剂,并用RT-PCR和Western Blot方法对筛选得到的小分子化合物进行验证。利用免疫印迹法分析筛选得到的化合物对细胞内信号通路的影响。结果通过筛选发现大豆甙元能够激活ChAT启动子的活性,与对照相比具有显著性差异(P<0.05)。进一步研究结果显示,大豆甙元也可以促进ChAT mRNA和蛋白的表达。另外,大豆甙元能够促进细胞内ERK的磷酸化,并进而激活ERK/MAPK信号通路。结论大豆甙元能够促进ChAT基因的表达,并引起细胞内ERK/MAPK信号通路的改变。
Objective Screening of ChAT gene expression activator from traditional Chinese medicine compounds li-brary.Methods Screening of ChAT gene expression activator from over 400 compounds by using ChAT gene promoter driven luciferase reporter system,then investigated the compound effect on the expression of ChAT mRNA and pretein by RT-PCR and western blot.The effect of compounds on intracellular signaling pathway was study by western blot. Results Daidzein was found that can promote the activity of ChAT promoter （P〈0.05 ）,and further increase the ChAT expression in both mRNA and protein levels.In addition,Daidzein can promote ERK1/2 phosphorylation and ac-tivite the ERK/MAPK signaling pathway.Conclusion Daidzein can promote the expression of ChAT gene,and activite the ERK/MAPK signaling pathway.
出处
《中国实验诊断学》
2014年第10期1576-1578,共3页
Chinese Journal of Laboratory Diagnosis
