药物洗脱法模拟阿片类药物戒断细胞模型的建立

目的 应用药物急性洗脱法建立阿片类药物非纳洛酮催促戒断细胞模型.方法 以稳定表达μ阿片受体(Mu opioid receptor,MOR)的CHO细胞CHO-MOR为研究对象,阿片类药物(D-Ala2,N-甲基-Phe,Gly-ol)脑啡肽(DAMGO)及吗啡预处理细胞不同时间,然后采用药物洗脱法将药物去除,运用cAM...

Establishment of drug-eluting simulated opioids withdrawal cell model

Objective To establish a kind of cell model of drug-eluting simulated opioids withdrawal, which distinguished from naloxone-precipitated withdrawal. Methods CHO-mu opioid receptor(MOR) cells which stablely express MOR were pretreated with （D-Ala2, N-Me-Phe, Gly-ol）-enkephalin （DAMGO） and morphine, then drugs were removed by drug-eluting. Using cAMP HTRF analysis detects the concentration of the second messenger cAMP in the down-stream pathways of MOR. The withdrawal index was cAMP overshooting. Results CHO-MOR cells were pretreated for 7.5 min, 15 min, 30 min, 1 h, 2 h, 3 h, 4 h with DAMGO, after removing drug by washing, the concentration of cAMP accumulated gradually with the prolonged DAMGO pretreatment. When CHO-MOR cells were exposed to DAMGO for 3 h and 4 h, it represented a significant rise in the level of cAMP-cAMP overshoot. Morphine pretreated CHO-MOR for 1 h, there was no marked changes in the concentration of cAMP. However, the level of cAMP was significantly higher than that of control group after 24 h of morphine pretreatment, which cloesd to that by naloxone-precipitated withdrawal. Conclusion The model of drug-eluting could better induce cAMP overshoot, which is an indication of the opioids withdrawal action in cells.