目的：利用干细胞构建可用于药物筛选的细胞模型，初步建立寻找中药活性成分的新方法。方法：首先，克隆神经生长因子（NGF）启动子的序列，连接到含有荧光素酶报告基因/E2红色荧光蛋白的质粒上，并将合成得到的质粒转入HEK-293细胞中，构建基于启动子的初步筛选稳定的细胞模型。使用阳性药进行验证细胞模型的准确性，并用该模型筛选来自中药的小分子化合物。然后使用筛选得到的化合物诱导大鼠的骨髓间充质干细胞，检测相关神经营养因子的转录水平，验证待测小分子化合物可能对神经生长的生物活性。结果：来自中药益智的三种化合物，白杨素、圆柚酮和杨芽黄素均可以激活NGF启动子。白杨素、圆柚酮诱导MSC不同时间后，细胞表面神经标志物:神经生长因子（NGF）、半乳糖神经酰胺（GalC）、巢蛋白（Nestin）、神经元特异烯醇化酶（NSE）、波形蛋白（Vimentin）对应RNA的转录水平均有不同程度的提高。而杨芽黄素没有明确促进神经细胞分化的活性。结论：白杨素、圆柚酮可以促进干细胞向神经细胞分化，是中药益智发挥神经保护作用物质基础的组成部分。

Objective: To construct a cell model that can be used for drug screening by using stem cells, and initially establish a new method for finding active ingredients of traditional Chinese medicines. Methods: First, the sequence of the nerve growth factor (NGF) promoter was cloned, ligated into the plasmid containing the luciferase reporter gene/E2red fluorescent protein, and the synthetic plasmid was transferred into HEK-293 cells, and the construction was startedbased on a preliminary screening of stable cell models. The positive drug was used to verify the accuracy of the cellmodel, and the model was used to screen small molecule compounds from Chinese medicine. Then, the obtainedcompound was used to induce bone marrow mesenchymal stem cells of the rat, and the transcription level of the relevantneurotrophic factor was detected to verify the biological activity of the small molecule compound to be tested for nervegrowth. Results: Three compounds from Chinese medicine Fructus Alpiniae Oxyphyllae, chrysin, nootkatone and popidin,activated the NGF promoter. Cellulose neuronal markers: nerve growth factor (NGF), galactosylceramide (GalC), nestin(Nestin), neuron-specific enolase (NSE), waveforms after induction of MSCs at different times in MSCs. The transcriptionlevel of the protein (Vimentin) corresponded to different degrees of RNA. However, popoflavin did not clearly promote theactivity of neural cell differentiation. Conclusion: Chrysin and nootkatone can promote the differentiation of stem cellsinto neural cells, which is a component of the neuroprotective effect of traditional Chinese medicine.

国家科技部重点研发计划（2018YFC1706200）：中医药现代化研究“常用中药活性成分的合成生物学研究，负责人：谭仁祥；四川省中医药管理局建设专项（2018C031）：中藏药大健康产品开发创新团队，负责人：谭睿。