cAMP反应元件结合蛋白在海洛因依赖大鼠杏仁核及海马的表达

Expression of cAMP response element binding protein in the amygdaloid and hippocampus of heroin-dependent rats

目的:观察cAMP反应元件结合蛋白在海洛因依赖SD大鼠杏仁核及海马的表达。方法:实验于2004-12在贵阳医学院生理教研室完成。选择健康SD大鼠20只,随机分成海洛因依赖组和生理盐水对照组,每组10只。海洛因(纯度61.48%,贵州省公安厅提供)。海洛因依赖组按剂量逐日递增原则,2次/d,连续9d皮下注射海洛因。对照组以同样方法注射等量生理盐水。第10天用纳洛酮催促戒断,确定海洛因依赖模型的建立。采用SABC免疫组化染色观察两组杏仁和海马cAMP反应元件结合蛋白的表达情况,结合计算机图像分析系统,比较两组杏仁和海马cAMP反应元件结合蛋白阳性细胞数和平均吸光度值。结果:纳入大鼠20只,实验组在建立模型过程中有1只死亡,另外随机选取大鼠补充,最终进入结果分析大鼠保持为20只。①SABC免疫组化染色发现,海洛因依赖组大鼠杏仁核cAMP反应元件结合蛋白阳性细胞数比对照组显著增多[(7.69±0.86),(6.66±1.23),P<0.05]。②两组大鼠杏仁核cAMP反应元件结合蛋白阳性细胞平均吸光度接近,差异无显著性[(215.56±4.17),(215.23±1.68),P>0.05]。③海洛因依赖组大鼠海马cAMP反应元件结合蛋白阳性细胞平均吸光度显著低于对照组,差异有显著性[(218.84±2.54),(222.27±2.67),P<0.05]。④两组大鼠海马cAMP反应元件结合蛋白阳性细胞数接近,差异无显著性[(17.34±1.90),(17.66±1.53),P>0.05]。结论:cAMP反应元件结合蛋白在海洛因依赖大鼠的杏仁核及海马的表达增强,提示cAMP反应元件结合蛋白参与海洛因依赖的形成,可能与成瘾记忆有关。

AIM： To observe the expression of cAMP response element binding protein （CREB） in the amygdoid and hippocampus of heroin-dependent rats. METHODS： The experiment was carried out in the Department of Physiology, Guiyang Medical College in December 2004. Twenty healthy SD rats were randomly divided into heroin-dependent group （n=10） and saline control group （n=10）. Heroin with the purity of 61.48% was provided by Guizhou Provincial Police Bureau. Rats in the heroin-dependent group and saline control group were given subcutaneous injection of heroin and saline respectively with the dosage increased day by day, twice a day for 9 continuous days. On the 10th day, the withdrawal signs precipitated by intraperitoneal injection of naloxone were evaluated. SABC immunohistochemical methods was applied to observe the expression of CREB in the amygdoid and hippocampus of rats in both groups. The number and mean absorbance value of CREB positive cells were compared between the two groups by using the image analysis. RESULTS： Totally 20 rats were used, 1 in the experimental group died during the experiment, and then another 1 was randomly selected for complement, finally 20 rats were involved in the analysis of results. ① SABC immunohistochemical staining showed that the number of CREB positive cells in amygdoid in the heroin-dependent group was significantly increased as compared with that in the control group [（7.69±0.86）, （6.66± 1.23）, P 〈 0.05].② The mean absorbance value of CREB positive cells in amygdoid was insignificantly different between the two groups （215.56±4.17, 215.23±1.68, P 〉 0.05）. ③ The mean absorbance value of CREB positive cells in hippocampus in the experimental group was significantly lower than that in the control group （218.84±2.54, 222.27±2.67, P〈 0.05）. ④ The number of CREB positive cells in hippoeampus was close between the two groups with insignificant difference （17.34 ± 1.90, 17.66 ± 1.53, P 〉 0.05）. CONCLUSION： The expression of CREB was enhanced in the amygdoid and hippoeampus of heroin-dependent rats, it is indicated that CREB is involved in the formation of heroin-dependence, which is associated with addicted memory.