镇心省睡益智方及其挥发油对记忆损伤小鼠的保护作用研究

Protective Effect of ZXXSYZ Fang and Its Essential Oil on Learning Ability and Memory of Alzheimer’s Disease Model Mice

目的:观察镇心省睡益智方(省睡方)水提液及其省睡方药对益智-石菖蒲挥发油对Aβ1-42诱导阿尔茨海默病(AD)小鼠学习记忆及神经递质的影响。方法:运用ICR小鼠侧脑室注入Aβ1-42复制记忆损伤模型小鼠,将省睡方水提液(0.455 g/kg)进行灌胃给药,不同剂量益智-石菖蒲挥发油(12.13μg/mL、48.50μg/mL)进行吸嗅给药,采用跳台实验、Morris水迷宫实验对小鼠行为学能力进行检测,采用尼氏染色光镜下观察海马CA1区神经元变化,运用Elisa法检测小鼠海马组织中乙酰胆碱(Ach)、伽马氨基丁酸(GABA)、谷氨酸(Glu)含量的变化。结果:跳台实验中,与空白组相比,模型组跳台实验潜伏期明显缩短、错误次数明显增加;与模型组相比,各给药组均可以显著延长小鼠跳台实验潜伏期、减少跳台错误次数;Morris水迷宫定位航行实验显示,模型组小鼠基本丧失学习记忆能力;各给药组与第一天相比,逃避潜伏期明显减少;空间探索实验显示各组之间无显著差异;在尼氏染色中,镇心省睡益智方灌胃给药和益智-石菖蒲挥发油吸嗅给药均可以一定程度保护小鼠海马CA1区神经元;Elisa结果显示,与空白组相比,模型组小鼠海马组织中Ach含量显著降低,GABA、Glu含量显著性升高;与模型组相比,镇心省睡益智方灌胃0.455 g/kg组及益智-石菖蒲挥发油吸嗅给药12.13μg/mL、48.50μg/mL组小鼠海马组织中Ach含量明显升高,GABA、Glu含量明显降低。结论:镇心省睡益智方水提液及挥发油灌胃及吸嗅给药可以改善Aβ1-42所致AD模型小鼠的学习记忆行为、保护神经元,升高Ach含量,降低GABA、Glu含量可能是其防治AD的机制之一。

Objective:To evaluate the protective effects of water extract from ZHENXIN XINGSHUI YIZHI FANG(ZXXSYZF)and the essential oil from Alpinia oxyphylla and rhizoma acori graminei on learning memory and neurotransmitters of Aβ1-42 induced AD model mice.Methods:ICR mice were intracerebroventricularlly injected with Aβ1-42 to establish the Alzheimer’s Disease(AD)mice model.0.455 g/kg ZXXSYZF water extract was orally administered,and different doses of essential oil(12.13μg/mL and 48.50μg/mL)were administered as inhalation.The Platform and Morris water maze tests were used to determine the mice behavior ability.Changes of brain ultrastructure(CA1)were observed by optical microscopy(Nissl stain).Contents of neurotransmitters Ach,Glu and GABA in hippocampus of AD mice were determined by ELISA.Results:In the platform test,compared with the control group,the latency period was significantly shortened and the error number were significantly increased in the model group.Compared with the model group,all doses of drug groups significantly prolonged the latency and reduced the error numbers.In Morris water maze tests,mice showed little learning and memory function in the model group.In other drug groups,compared with the first day,the escape latencies were significantly reduced,but space exploring abilities showed no significant difference.In the Nissl staining,the protective effects on hippocampus CA1 neurons were observed in both ZXXSYZF water extract group and essential oil group.In ELISA test,compared with the control group,in the model group,the level of Ach was significantly reduced and the levels of Glu and GABA were significantly increased.Compared with the model group,the levels of Ach were significantly increased and the levels of Glu and GABA were significantly reduced in 0.455 g/kg ZXSSYZF water extract group and essential oil groups(12.13μg/mL and 48.50μg/mL).Conclusion:The extracted solution from ZXSSYZF and essential oil can improve the learning memory and protect neurotransmitters in Aβ1-42 induced AD mice,its mechanism may be related to increasing the level of Ach and reducing the levels of Glu and GABA.