基于联合式学习机制建立小鼠海洛因成瘾行为学模型

A behavioral model of heroin addiction in mice based on associative learning mechanism

目的基于联合式学习机制建立一个实验室C57BL/6小鼠海洛因成瘾模型。方法选择黑箱与白箱作为记忆训练环境,开展3种行为学训练范式研究:①巴甫洛夫条件位置偏好(CPP)训练范式,上午9:00腹腔注射0.1 ml对应浓度(5.0、10.0、20.0 mg/kg)海洛因后将小鼠放入白箱进行记忆强化训练30 min,24 h后腹腔注射0.1 ml 0.9%氯化钠溶液放入黑箱训练,训练结束后检测小鼠对黑白箱的记忆偏好程度(不同箱体的运动时间)。②基于CPP训练范式的结果进行纳洛酮条件位置厌恶(CPA)训练范式;③行为敏化训练范式,基于3个行为学实验的统计学结果和实验小鼠给药后致死率建立了海洛因成瘾评分量表。选用3个不同剂量的海洛因(5.0、10.0、20.0 mg/kg)诱导海洛因成瘾,通过评分表上的结果来选择最合适的海洛因浓度。结果在CPP训练范式中,5.0、10.0、20.0 mg/kg海洛因组均出现CPP(P<0.05,P<0.001,P<0.05);CPA训练范式中,与对照组相比,10.0 mg/kg海洛因组CPA诱导率最高且2次诱导后出现CPA(P<0.01);行为敏化训练范式中,各浓度海洛因组均会引起行为敏化改变(P<0.001);其中5.0、10.0 mg/kg海洛因组不会导致动物死亡,10.0 mg/kg海洛因组的剂量在评分量表中得分最高,可作为建立稳定的海洛因成瘾实验动物模型浓度。结论该研究有效地建立起小鼠海洛因成瘾模型,且适用于建模药物浓度筛选,动物成活率高,简单实用。联合式学习机制能有效地缩短海洛因成瘾实验动物模型建立周期。

Objective To establish a laboratory model of heroin addiction in C57BL/6 mice based on associative learning mechanisms.Methods The black box and white box were selected as the memory training environment,and three behavioral training paradigms were studied:①Pavlovian conditional position preference(CPP)training paradigm,mice were placed in the white box for memory reinforcement training for 30 min after intraperitoneal injection of 0.1 ml of corresponding concentrations(5.0,10.0,20.0 mg/kg)of heroin at 9:00 a.m.,and 24 h later 0.1 ml of 0.9%NaCl solution was injected intraperitoneally into the black box for training,and after the training,the mice were tested for their memory preference for the black and white boxes(movement time of different boxes).②A naloxone conditional position aversion(CPA)training paradigm was conductedbased on the results of the CPP training paradigm.③Behavioral sensitization training paradigm,heroin addiction rating scale was established based on the statistical results of 3 behavioral experiments and the lethality of experimental animal disease mice after drug administration.Three different doses of heroin(5.0,10.0,20.0 mg/kg)were selected to induce heroin addiction,and the most appropriate heroin concentration was selected by the results on the rating scale.Results In the CPP training paradigm,CPP was observed in all heroin groups(P<0.05,P<0.001,P<0.05).In the CPA training paradigm,the CPA induction rate was highest in the 10.0 mg/kg heroin group compared to the control group(P<0.01).In the behavioral sensitization training paradigm,all heroin groups caused behavioral sensitization changes(P<0.001);but the 5.0 and 10.0 mg/kg heroin groups did not cause animal mortality.Overall,the 10.0 mg/kg heroin group had the highest dose score on the rating scale.It could be used as a concentration to establish a stable experimental animal model of heroin addiction.Conclusion The study was effective in establishing a heroin addiction model in mice,and it was suitable for modeling drug concentration screening,with high animal survival rate and simple and practical.The combined learning mechanism can effectively shorten the model establishment period.