应用放射性免疫方法建立抗早老性痴呆药的M1受体筛选模型

目的 :建立M1受体筛选模型 ,用于抗早老性痴呆药物的筛选。方法 :在人胚肾细胞上表达人源M1受体 ,通过检测胞内cAMP来监测受体的激动情况。使用外源性的3H cAMP竞争胞内的cAMP结合位点。使用氯化乙酰胆碱作为阳性药来确证模型的可靠性。结果 :初步建立了M1受体人胚肾细胞表达体系。使用不同浓度的氯化乙酰胆碱 (10 -9～ 10 -4mol/L) ,胞内的cAMP浓度有相关性变化 (10 3 43× 10 -4 ～ 3 3 75 4× 10 -4 pmol/ μl)。 结论 :本筛选模型对M1受体激动剂有阳性反应 ,可用于抗早老性痴呆药物的筛选

Establishment of M1 Receptor Screening Model for Drugs for Alzheimer's Disease Using Radioimmunoassay Method

AIM:To establish a screening model based on muscarinic receptor type 1(M1) for drugs against Alzheimer's disease (AD). METHOD: Human M1 receptor was expressed in HEK293 (human embryonic kidney) cells, and its activation was measured by the intracellular cAMP (cyclic AMP) level. Exogeneous 3H-cAMP was used to compete the intracellular cAMP binding sites. Acetylcholine chloride was used as a positive drug to ensure the sensitivity of this model. RESULT: HEK293 cell expressing system of human M1 receptor was established. Different concentrations of acetylcholine chloride (10-9～10-4 mol/L) activation of M1 receptor leads to an increase of intracellular cAMP 10.343×10-4～33.754×10-4 pmol/μl. CONCLUTION:This screening model has positive response to M1 receptor agonist and can be used for drug screening.