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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 cell model and method for screening Cysteinyl Leukotriene Receptor 2 (CysLT2 receptor) antagonists and to screen a series of synthetic compounds. Methods: Rat C6 glioma cells with high expression of CysLT2 receptor were selected and the intracellular calcium concentration was measured after LTD4 challenge. As a screening method of CysLT2 antagonist, compounds with antagonist activity were screened. CysLT1 / CysLT2 receptor non-selective antagonist Bay u9773 and CysLT2 receptor selective antagonist AP-100984 as a reference. Results: RT-PCR showed that C6 cells highly expressed CysLT2 receptor. LTD41μmol / L could significantly increase intracellular calcium in C6 cells, which was about 37.5% of that of positive control ATP. CysLT2 receptor antagonist inhibits LTD4-induced intracellular calcium elevation in C6 cells, and CysLT1 receptor antagonists have no inhibitory effect. DXW-1, 2, 13, 23, 29, and 30 in compounds inhibit LTD4-induced intracellular calcium elevation. CONCLUSION: The changes of calcium signaling in C6 cells induced by LTD4 can be used as a screening method for CysLT2 receptor antagonists. Compounds DXW-1, 2, 13, 23, 29 and 30 have CysLT2 receptor antagonistic activity.