卤素离子通道阻断剂的高通量筛选

High-throughput Screening of Halogen Ion Channel Blockers

【目的】从天然化合物中初步筛选出具有阻断细胞膜上阴离子（卤素）通道的候选化合物，为新药研发提供实验依据。【方法】利用前期实验已构建EYFP-H148Q／1152L-HeLa细胞系为模型，在已知结构及来源的6988种天然化合物单体库中，通过应用低渗诱导细胞阴离子通道开放，是否发生荧光的淬灭来筛选出具有阻断阴离子（卤素）通道的化合物。【结果】①Ⅰ-（低渗）溶液组为阴性对照，细胞荧光淬灭。②Cl-（等渗）溶液组为阳性对照，细胞荧光强度不变。③Cl-离子通道阻断剂DIDS具有阻断阴离子通道作用，以细胞荧光淬灭强度下降50％以内，以DIDS为筛查参考标准。【结论】低渗刺激EYFP-H1480／1152L-HeLa细胞模型阴离子（卤素）通道开放，荧光淬灭，载体所表达YFP具有活性功能。利用该模型在6988种化合物单体库中初步筛选出7种候选化合物具有阻断细胞膜上阴离子通道的作用。

[Objective] To screen out the candidate compounds capable of blocking cell membrane anion （halogen） channel from the natural compounds so as to provide reference for the research and development of new drugs.[Methods]The EYFP-H148Q/I152L-HeLa cell line model was established by previous experi- ments. In the monomer library of 6988 natural compounds of known structure and origin, the compounds with blocking anion （halogen） channel were screened out through the application of the channel opening of low per- meability induced cell anion and the quenching degree of fluorescence.[Results]（2）Ⅰ- （low permeability） solu- tion as negative control group, cell fluorescence quenched. （2）C1-（isotonic） solution as positive control, cell fluorescence intensity was constant.（3）Cl- ion channel blocker DIDS have blocking anion channel effect, cell flu- orescence quenched below 50%, DIDS were set as the standard of drug screening. [Conclusion] In EYFP- H148Q/I152L-HeLa cell model with low permeability, the anion （halogen） channel was opened, and fluores- cence quenched; and the YFP expressed by the carrier has the function of activity. With the natural monomers extracted in Rauvolfia, Sauussurea Costus, Zingiberaceae and Aloe,7 candidate compounds were screened out, which are able to block the anion channel on the cell membrane.By using the model, 7 kinds of the initial candi- date compounds were screened out from 6988 natural compounds, which are able to block the anion channel on the cell membrane.