黑逍遥散对AD模型小鼠海马区APP,PERK表达的影响

Effect of Hei Xiaoyaosan on Expressions of APP,PERK in Hippocampus Area of Alzheimer’s Dementia Mice

目的:研究黑逍遥散对阿尔茨海默症(AD)模型小鼠海马神经元内质网应激的影响,包括行为学、组织病理学及淀粉样前体蛋白(APP),类蛋白激酶内质网激酶(PERK)等因子的表达。方法:使用SPF级的4月龄双转基因(APP/PS1)雄性小鼠42只随机分为黑逍遥散高、低剂量组(6,3 g·kg-1),盐酸多奈哌齐组(3. 25 mg·kg-1)及模型组4组,同种系同月龄C57BL小鼠10只作为正常组。首先适应环境1周,经不同药物干预治疗2个月后,用Morris水迷宫行为学检测每组小鼠的学习和记忆能力;再治疗1个月后,通过光镜来观察每组小鼠海马区组织病理学改变;免疫组化、实时荧光定量聚合酶链式反应(Real-time PCR)检测小鼠海马区细胞内质网组织中APP,PERK蛋白及mRNA的表达情况。结果:药物干预治疗后,与正常组比较,模型组小鼠的逃避潜伏期显著延长(P <0. 01);模型组小鼠海马区内神经元严重损伤;模型组APP,PERK的表达量显著升高(P <0. 01)。与模型组比较,各给药组小鼠逃逸潜伏期均明显缩短(P <0. 05,P <0. 01);海马区内神经元损伤程度均减轻;APP,PERK的表达均明显降低(P <0. 05)。结论:黑逍遥散能够显著改善AD小鼠的学习记忆能力,可能与减少内质网过度应激反应等方面来减轻AD小鼠认知能力损伤有关。

Objective:To study the effect of Hei Xiaoyaosan on endoplasmic reticulum stress in hippocampal neurons of Alzheimer’s disease(AD)model mice,including behavioral,histopathology and amyloid precursor protein(APP),protein kinase endoplasmic reticulum kinase(PERK)expressions.Method:The 42 4-month-old SPF-grade double transgenic(APP/PS1)mice were randomly divided into the high-dose group and the low-dose group,the donepezil hydrochloride group and the model group,and 10 C57BL mice of the same age were used as the blank group.Firstly,they were adapted to the environment for one week.After 2 months of treatment with different drug interventions,Morris water maze behavior was used to test the learning and memory abilities of each group of mice.After 1 month of treatment,histopathological changes in the hippocampus of each group of mice were observed by light microscopy.The expressions of APP,PERK protein and mRNA in the endoplasmic reticulum of hippocampus were detected by immunohistochemistry(IHC)and real-time fluorescent quantitative polymerase chain reaction(Real-time PCR).Result:After drug intervention,compared with the blank group,the escape latency of the AD model group was significantly prolonged(P<0.01),the neurons in the hippocampus of the model group were severely damaged,and the expressions of APP and PERK in the model group were significantly increased(P<0.01).In the comparison model group,the escape latency of the mice in each treatment group was significantly shortened(P<0.05,P<0.01),the degree of neuronal damage in the hippocampus of each treatment group was alleviated,and the positive expressions of APP and PERK in each treatment group were weakened(P<0.05).Conclusion:Hei Xiaoyaosan can significantly improve the learning and memory abilities of AD mice,which may be related to the reduction of the excessive stress response of endoplasmic reticulum to alleviate cognitive impairment in AD mice.