P54/cAMP/PKA信号调控在酒精依赖大鼠中的相关性研究

Correlation study of P54/cAMP/PKA signal regulation in ethanol dependent rats

目的 探讨P54/cAMP/PKA信号调控在酒精依赖大鼠中的相关性。方法 使用随机数表法将SD大鼠(n=50,雄性)随机分为对照组(control组,n=10)、10%双瓶自由连续性饮酒模型组(10%ethanol组,n=10)、慢病毒阴性对照组(JL-control组,n=10)、慢病毒空载组(JL-NC组,n=10)和P54过表达组(JL-P54组,n=10),在构建模型28 d后,JL-control组、JL-NC组和JL-P54组进行转染。取五组大鼠海马组织进行Western Blot检测和实时荧光定量PCR(qRT-PCR)检测。结果 10%ethanol组在最后7 d的酒精摄入量(16.80±0.63)ml/24 h保持稳定,饮水量(11.90±0.39)ml/24 h保持稳定,酒精偏好稳定在(57.74±1.83)%,酒精摄入量逐渐增加到达平台期,饮水量伴随饮酒量的增多而降低到稳定,酒精偏好增加到稳定;在Western Blot检测和qRT-PCR检测中,与control组比较,10%ethanol组中P54、腺苷酸环化酶(AC)、cAMP依赖性蛋白激酶(PKA)、cAMP反应元件结合蛋白(CREB)的表达量下降,差异有统计学意义(P <0.05),JL-P54组中P54、AC、PKA、CREB的表达量升高,差异有统计学意义(P <0.05)。结论 P54可能通过cAMP/PKA信号通路的调控参与酒精依赖的形成机制。

Objective To explore the correlation of P54/cAMP/PKA signal regulation in ethanol dependent rats.Methods SD rats(n=50,male)were randomly divided into a control group(n=10),a 10%two-bottle free continuous drinking model group(the 10%ethanol group,n=10),a lentivirus negative control group(the JL-control group,n=10),a lentivirus no-load group(the JL-NC group,n=10)and a P54 overexpression group(the JL-P54 group,n=10)by a random number table method.After 28 days of model construction,the JL-control group,JL-NC group and JL-P54 group were transfected.Hippocampal tissues of the five groups were taken for the Western Blot test and quantitative real-time PCR(qRT-PCR)test.Results The ethanol intake of the 10%ethanol group remained stable at(16.80±0.63)ml/24 hours in the last 7 days,and the water intake remained stable at(11.90±0.39)ml/24 hours.The ethanol preference remained stable at(57.74±1.83)%,and the ethanol intake gradually increased to a plateau.The water intake decreased to a stable level with the increase in ethanol consumption,and the ethanol preference increased to a stable level;In the Western Blot test and qRT-PCR test,compared with the control group,the expressions of P54,adenylate cyclase(AC),cAMP-dependent protein kinase(PKA),cAMP response element binding protein(CREB)in the 10%ethanol group decreased,with statistically significant differences(P<0.05);The expressions of P54,AC,PKA and CREB in the JL-P54 group increased,with statistically significant differences(P<0.05).Conclusion P54 may participate in the formation mechanism of ethanol dependence through the regulation of cAMP/PKA signaling pathway.