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柠檬酸转运蛋白(Na CT)为新发现的钠离子偶联的羧酸转运蛋白家族的一员,主要功能是负责转运三羧酸循环的中间体,近年来研究发现Na CT可能是治疗糖尿病和肥胖症的重要药物靶点。该研究构建了Na CT基因的真核表达质粒,通过稳定转染HEK293细胞,筛选得到了稳定表达Na CT的细胞系。利用Rapid Fire平台建立了基于Na CT的高通量药物筛选细胞模型,结果显示该模型能高效地转运柠檬酸,具有良好底物特异性和选择性,其转运能力可被Li+激活,被4,4-二异硫氰基芪-2,2’-二磺酸抑制。Z因子计算结果为0.69,提示其适用于高通量筛选。对两种不同类型化合物库的筛选获得了一系列Na CT抑制剂,包括14个小分子化合物和15个中药组分。基于Na CT细胞模型的成功构建为高通量筛选具有抗糖尿病及肥胖症作用的先导化合物提供了一个新平台。
Citrate transporter (Na CT) is a member of the newly discovered family of Na + -coupled carboxyfusate transporters. Its main function is the intermediate responsible for transporting the tricarboxylic acid cycle. In recent years, it has been found that Na CT may be used to treat diabetes and Obesity is an important drug target. In this study, an eukaryotic expression plasmid of Na CT gene was constructed and stably transfected into HEK293 cells. The cell lines stably expressing Na CT were screened out. The Rapid Fire platform was used to establish a high-throughput NaCN-based drug screening cell model. The results showed that this model can efficiently transport citric acid with good substrate specificity and selectivity. Its transport capacity can be activated by Li + 4-diisothiocyanatostilbene-2,2’-disulfonic acid inhibition. The Z factor was calculated to be 0.69, suggesting that it is suitable for high-throughput screening. Screening of two different types of compound libraries obtained a series of Na CT inhibitors, including 14 small molecule compounds and 15 Chinese herbal components. The successful construction based on Na CT cell model provides a new platform for high-throughput screening of lead compounds with anti-diabetic and obesity effects.