清心开窍方对APP/PS1双转基因小鼠海马区AKT/GSK3α/βapp/Aβ表达的影响

Effect of Qingxin Kaiqiao Recipe on Expressions of AKT,GSK3α,βAPP and Aβ in Hippocampus of APP/PS1 Transgenic Mice

目的：探讨清心开窍方对APP/PS1双转基因小鼠海马AKT（protein kinase B,PKB,Akt）GSK3α（Glycogen synthase kinase 3α）、βAPP（β-amyloid precursor protein,βAPP）、Aβ（β-amyloid,Aβ）表达的影响。方法：选取60只APP/PS1双转基因小鼠,按随机数字表法随机分为模型组、多奈哌齐组（1.67 mg/kg）、清心开窍方高剂量组（19 mg·kg-1·d-1）,清心开窍方中剂量组（9.5 mg·kg-1·d-1）,清心开窍方低剂量组（4.75 mg·kg-1·d-1）,另选取同背景同性别C57BL/6J小鼠为对照组,对照组、模型组给予等量生理盐水灌胃,各组每天干预1次,连续灌胃12周。RT-PCR检测AKT、GSK3α、βAPP、AβmRNA的表达,Western Blot法检测AKT、GSK3α、βAPP、Aβ蛋白表达量,硫磺素T染色法测定小鼠大脑皮层及海马区老年斑的数量。结果：实时荧光定量PCR和Western Blot结果显示,模型组中GSK3α、βAPP、AβmRNA的表达最高,与对照组及其他各组相比有统计学意义（P〈0.05或P〈0.01）,且其表达量随清心开窍方剂量的增加而减少;模型组中AKT的表达量最低,与对照组及其他各组相比有统计学意义（P〈0.05或P〈0.01）,且其表达量随清心开窍方剂量的增加而增加。硫磺素T染色结果显示,模型组中老年斑数量最多,与对照组及其他各组相比有统计学意义（P〈0.05或P〈0.01）。结论：清心开窍方能够促进APP/PS1双转基因小鼠脑内AKT的表达,并减少GSK3α、βAPP和Aβ的表达,减少APP/PS1双转基因小鼠脑内皮质及海马区的老年斑数量。

Abstract：Objective： To investigate the effect of Qingxin Kaiqiao Recipe on the expressions of AKT,GSK3α, βAPP and Aβ in hippocampus of APP/PS1 transgenic mice. Methods： A total of 60 APP/PS1 transgenic mice were randomly divided into model group, Donay Pizzigo （ 1.67 mg/kg）, Qingxin Kaiqiao Recipe （ 19 mg · kg^-1 · d^-1） high dose group, Qingxin Kaiqiao Recipe low dose group（9.5 mg/ · kg^-1 · d^-1）, Qingxin Kaiqiao Recipe low dose group（4.75 mg · kg^-1 · d^-1 ）. The C57BL/6J mice were used as control group. The control group and model group were given normal saline. Each group was intervened once a clay,lasting for 12 weeks. RT- PCR was used to detect the expressions of AKT, GSK3α, βAPP and Aβ, and the expressions of AKT, GSK3α, βAPP and Aβ proteins were detected by Western Blot. The number of senile plaques in the cerebral cortex and hippocampus of mice were measured by sulfur T staining method. Results： RT- PCR and Western Blot results showed compared with the control group and the drug groups, the model group had the highest contents of GSK3α, βAPP and Aβ and the lowest contents of AKT. There was a statistically significant difference （P 〈 0. 05 or P 〈 0. 01 ） between them and their contents had a correspondence with the dosage of Qingxin Kaiqiao Recipe. The results of sulfur T staining showed that the number of senile plaques in the model group was the highest compared with that in the control group and other groups （P 〈 0. 05 or P 〈 0.01 ）. Conclusion：Qingxin Kaiqiao Recipe can promote the expression of AKT and reduce the contents of GSK3α, βAPP and Aβ in the brain of APP/PS1 transgenic mice. It also can reduce the number of senile plaques in the cortex and hippocampus of APP/PS1 transgenic mice.