黑逍遥散对APP/PS1双转基因小鼠海马CREB1蛋白及其磷酸化表达的影响

Effect of Heixiaoyao Powder(黑逍遥散) on CREB1 Protein and Its Phosphorylation Expression in Hippocampus of APP/PS1 Double Transgenic Mice

目的探讨黑逍遥散治疗阿尔茨海默病的可能作用机制。方法 30只APP/PS1双转基因雄性小鼠随机分为模型组、盐酸多奈哌齐组、黑逍遥散组,每组10只。同月龄、同种系的野生型C57BL/6雄性小鼠10只设为空白组。黑逍遥散组给予黑逍遥散6 g/(kg·d)灌胃,盐酸多奈哌齐组给予盐酸多奈哌齐片3. 25 mg/(kg·d)灌胃,模型组、空白组给予双蒸水0. 2 ml/10 g灌胃,各组均每日灌胃1次,连续3个月。采用Morris水迷宫于实验结束第1~5天进行定位航行实验观察逃避潜伏期,第6天开展空间搜索实验记录跨原平台次数。采用免疫组化法检测小鼠海马CA1、CA3及DG区环腺苷酸应答元件结合蛋白1 (CREB1)及磷酸化环腺苷酸应答元件结合蛋白1 (p-CREB1)表达。结果与空白组比较,模型组小鼠第1~5天逃避潜伏期明显延长,第6天跨原平台次数明显减少,小鼠海马CA1、CA3及DG区CREB1及p-CREB1蛋白表达显著降低(P <0. 05或P <0. 01)。与模型组比较,盐酸多奈哌齐组和黑逍遥散组各时间点逃避潜伏期均缩短,第6天跨原平台次数明显增加,小鼠海马CA1、CA3及DG区CREB1、p-CREB1蛋白表达显著升高(P <0. 05或P <0. 01)。黑逍遥散组和盐酸多奈哌齐组各指标组间比较差异无统计学意义(P> 0. 05)。结论黑逍遥散可能通过调节海马CREB1及其磷酸化表达发挥防治阿尔茨海默病的作用。

Objective To investigate the possible mechanism of action of Heixiaoyao Powder(黑逍遥散)in the treatment of Alzheimer’s disease.Methods A total of 30 APP/PS1 double transgenic male mice were randomly divided into a model group,a donepezil hydrochloride group and a Heixiaoyao Powder group,with 10 rats in each group.Ten wild-type C57 BL/6 male mice of the same age and same strain were set as blank group.The Heixiaoyao Powder group was given 6 g/(kg·d)by intragastric administration,and donepezil hydrochloride was given 3.25 mg/(kg·d)to the donepezil hydrochloride group.The model group and the blank group were given double distilled water 0.2 ml/10 g by gavage.All groups were intragastrically administered once a day for 3 months.The Morris water maze was used to observe the escape latency during the 1 st to 5 th day of the experiment,and the space search experiment was recorded on the 6 th day to record the number of times across the original platform.The expression of cyclic adenosine responsive element binding protein 1(CREB1)and phosphorylated cyclic adenosine responsive element binding protein 1(p-CREB1)in mouse hippocampal CA1,CA3 and DG regions were detected by immunohistochemistry.Results Compared with the blank group,the escape latency of the model group was significantly prolonged on the 1 st to 5 th day,and the number of trans-platforms was significantly decreased on the 6 th day.The expression of CREB1 and p-CREB1 protein in the hippocampus CA1,CA3 and DG areas was significantly decreased(P<0.05 or P<0.01).Compared with the model group,the escape latency of the donepezil hydrochloride group and the Heixiaoyao Powder group was shortened at each time point,and the number of trans-platforms increased significantly on the 6 th day.The expression of CREB1 and p-CREB1 protein in the hippocampus CA1,CA3 and DG areas of the mice increased significantly(P<0.05 or P<0.01).There was no significant difference between the indicators of Heixiaoyao Powder and Donepezil Hydrochloride(P>0.05).Conclusion Heixiaoyao Powder may play a role in the prevention and treatment of Alzheimer’s disease by regulating hippocampal CREB1 and its phosphorylation expression.