邻苯二甲酸二异丁酯——新型的ANO2通道开放剂

DIBP as a novel ligand for ANO2 channel activating

目的筛选ANO2的特异性配体药物,为医药行业开发靶向ANO2新药提供先导化合物的结构信息。方法构建ANO2过表达的HEK293细胞株,应用共聚焦显微镜-膜片钳全细胞记录模式技术筛选ANO2的特异性配体。结果在胞内外钳制电压100 mV及含1μmol·L^(-1)钙离子电极内液的记录条件下,邻苯二甲酸二异丁酯(diisobutyl phthalate,DIBP)给药前ANO2介导的氯电流密度为(14.67±2.87)pA/pF,给药后ANO2介导的氯电流电导为(28.75±4.05)pA/pF,差异具有统计学意义(P<0.01);钙荧光成像实验结果表明DIBP无调节胞内游离钙信号的作用;在0μmol·L^(-1)钙离子条件下,DIBP对静息状态的ANO2通道不具有直接的激活作用。结论DIBP通过直接作用增强已活化ANO2通道的电导,其作为一种新型的促ANO2通道开放剂为开发靶向ANO2的药物提供结构信息,同时也为研究其他邻苯二甲酸酯类物质的毒理作用提供一定的理论基础。

Aim To screen specific ligand for ANO2 to provide structural information of lead compounds for the development of new drugs targeting ANO2.Methods ANO2 overexpressed HEK293 cell line was constructed using liposome transfection,and the specific ligand of ANO2 was screened by confocal microscopy and patch-clamp technique.Results Under the recording conditions of transmembrane depolarized potential 100 mV and 1μmol·L^(-1) calcium pipette solution,ANO2-mediated chloride current Idensity was 14.67±2.87 pA/pF before DIBP administration and was 28.75±4.05 pA/pF after DIBP administration,which showed significant statistical difference.The results of calcium fluorescence imaging experiment showed that DIBP had no regulated effect on the cytoplasm calcium signaling.When using 0μmol·L^(-1) calcium pipette solution,DIBP could not direct activate ANO2 channel.Conclusions DIBP enhances the conductance of the opening ANO2 channel through direct action.As a novel ANO2 channel agonist,DIBP provides structural information for the development of ANO2-targeting drugs and provides strong theoretical basis for the study of the toxicological effects of other phthalates.