CAJ | 학술논문

目的:建立半胱氨酰白三烯受体2(CysLT_2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CysLT_2受体的大鼠C6胶质瘤细胞,激动剂LTD_4攻击后检测细胞内钙浓度变化,作为CysLT_2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/CysLT_2受体非选择性拮抗剂Bay u9773和CysLT_2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CysLT_2受体.LTD_4 1 μmol/L能显著升高C6细胞内钙,其作用约为阳性对照ATP的37 5%.CysLT_2受体拮抗剂抑制LTD_4诱导的C6细胞内钙升高,CysLT_1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LTD_4诱导的细胞内钙升高.结论:LTD_4诱导的C6细胞的钙信号变化可作为CysLT_2受体拮抗剂的筛选方法;化合物DXW-1、2、13、23、29、30具有CysLT_2受体拮抗活性.
목적:건립반광안선백삼희수체2(CysLT_2수체)길항제사선적세포모형급방법,초사계렬합성화합물.방법:선용고표체CysLT_2수체적대서C6효질류세포,격동제LTD_4공격후검측세포내개농도변화,작위CysLT_2수체길항제적사선방법,초보사선유길항제활성적화합물.채용CysLT1/CysLT_2수체비선택성길항제Bay u9773화CysLT_2수체선택성길항제AP-100984작위삼조.결과:RT-PCR감정표명C6세포고표체CysLT_2수체.LTD_4 1 μmol/L능현저승고C6세포내개,기작용약위양성대조ATP적37 5%.CysLT_2수체길항제억제LTD_4유도적C6세포내개승고,CysLT_1수체선택성길항제무억제작용.화합물중DXW-1、2、13、23、29、30능억제LTD_4유도적세포내개승고.결론:LTD_4유도적C6세포적개신호변화가작위CysLT_2수체길항제적사선방법;화합물DXW-1、2、13、23、29、30구유CysLT_2수체길항활성.
Objective: To establish a method for screening cysteinyl leukotriene receptor 2 (CysLT_2) antagonists and to preliminarily screen a series of synthetic compounds. Methods: Rat glioma cell line (C6 cells) highly expressing CysLT_2 receptor was used.Intracellular calcium concentration was measured after stimulation with the agonist LTD_4,which was used to screen compounds with antagonist activity for CysLT_2 receptor.Bay u9773,a CysLT_1/CysLT_2 receptor non-selective antagonist,and AP-100984,a CysLT_2 receptor antagonist,were used as control. Results: PT-PCR showed a higher expression of CysLT_2 receptor in C6 cells.LTD_4 at 1 μmol/L significantly increased intracellular calcium in C6 cells;the maximal effect was about 37.5% of ATP,a positive stimulus.LTD_4-induced increase of intracellular calcium was blocked by CysLT_2 receptor antagonists,but not by CysLT_1 receptor antagonists.Among the synthetic compounds,DXW-1,2,13,23,29 and 30 inhibited LTD_4-induced increase of intracellular calcium. Conclusions: LTD_4-induced change in intracellular calcium in C6 cells can be used as a screening method for CysLT_2 receptor antagonists.The compounds,DXW-1,2,13,23,29 and 30,possess antagonist activity for CysLT_2 receptor.