半胱氨酰白三烯受体2拮抗剂筛选方法的建立及化合物初筛  

Method for screening cysteinyl leukotriene receptor 2 antagonists and preliminary screening of compounds

作　　者：蔡蓓蕾[1] 黄雪琴[1] 董晓武[2] 方三华[1] 卢韵碧[1] 张纬萍[1] 胡永洲[2] 魏尔清[1] 

机构地区：[1]浙江大学医学院药理学系,浙江杭州310058 [2]浙江大学药学院药物研究所,浙江杭州310058

出　　处：《浙江大学学报（医学版）》2009年第6期598-604,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金资助项目(30772561);浙江省科技计划重点资助项目(2005C23041)

摘　　要：目的:建立半胱氨酰白三烯受体2(CysLT2受体)拮抗剂筛选的细胞模型及方法,初筛系列合成化合物。方法:选用高表达CysLT2受体的大鼠C6胶质瘤细胞,激动剂LTD4攻击后检测细胞内钙浓度变化,作为CysLT2受体拮抗剂的筛选方法,初步筛选有拮抗剂活性的化合物。采用CysLT1/CysLT2受体非选择性拮抗剂Bay u9773和CysLT2受体选择性拮抗剂AP-100984作为参照。结果:RT-PCR鉴定表明C6细胞高表达CysLT2受体。LTD41μmol/L能显著升高C6细胞内钙,其作用约为阳性对照ATP的37.5%。CysLT2受体拮抗剂抑制LTD4诱导的C6细胞内钙升高,CysLT1受体选择性拮抗剂无抑制作用。化合物中DXW-1、2、13、23、29、30能抑制LTD4诱导的细胞内钙升高。结论:LTD4诱导的C6细胞的钙信号变化可作为CysLT2受体拮抗剂的筛选方法;化合物DXW-1、2、13、23、29、30具有CysLT2受体拮抗活性。Objective： To establish a method for screening cysteinyl leukotriene receptor 2 （CysLT2） antagonists and to preliminarily screen a series of synthetic compounds. Methods： Rat glioma cell line （C6 ceils） highly expressing CysLT2 receptor was used. Intracetlular calcium concentration was measured after stimulation with the agonist LTD4,which was used to screen compounds with antagonist activity for CysLT2 receptor. Bay u9773,a CysLT1/CysLT2 receptor non selective antagonist, and AP 100984, a CysLT2 receptor antagonist, were used as control. Results ： PT-PCR showed a higher expression of CysLT2 receptor in C6 cells. LTD4 at 1μmol/L significantly increased intracellular calcium in C6 cells; the maximal effect was about 37.5% of ATP,a positive stimulus. LTD4 induced increase of intracellular calcium was blocked by CysLT2 receptor antagonists,but not by CysLT1 receptor antagonists. Among the synthetic compounds, Dxw-1,2,13,23,29 and 30 inhibited LTD4 induced increase of intracellular calcium. Conclusions ： LTD4-induced change in intracellular calcium in C6 cells can be used as a screening method for CysLT2 receptor antagonists. The compounds,Dxw 1,2, 13, 23, 29 and 30, possess antagonist activity for CysLT2 receptor.

关 键 词：半胱氨酸/遗传 受体 白三烯/遗传 半胱氨酰白三烯 CysLT2受体 受体拮抗剂 钙离子 C6细胞 

分 类 号：R96[医药卫生—药理学]