藏药旺拉活性成分对APP/PS1转基因5×FAD小鼠老年斑形成及炎症反应的影响

Effects of the Active Ingredients of Tibetan Medicine Wangla on Senile Plaques Formation and Inflammatory Response in APP/PS1 Transgenic 5×FAD Mice

目的:研究藏药旺拉活性成分对APP/PS1双转基因5×FAD小鼠脑内老年斑和炎症反应的影响。方法:选取20只4月龄5×FAD小鼠,随机分成模型组(n=10)和藏药旺拉活性成分组(n=10);20只同月龄野生型(WT)小鼠随机分为正常对照组(n=10)和藏药旺拉活性成分对照组(n=10)。正常对照组与模型组给予蒸馏水灌胃,藏药旺拉活性成分各组灌胃藏药旺拉活性成分。给药80 d后,应用Morris水迷宫实验检测各组学习记忆功能,免疫组化染色检测脑部淀粉样斑块沉积情况,免疫荧光染色和Western Blot观察脑部β-淀粉样蛋白(Aβ)沉积及离子钙结合接头分子1(Iba1)和胶质纤维酸性蛋白(GFAP)的表达分布并进行定量分析,ELISA检测脑内炎症因子水平。结果:与模型组比较,藏药旺拉活性成分组小鼠水迷宫潜伏时间显著减少,穿越目标象限的次数和停留时间显著增加,脑部淀粉样斑块沉积显著减少,Aβ、Iba1、GFAP表达及TNF-α、IL-6、IL-β水平显著降低(P<0.05~P<0.001)。结论:藏药旺拉活性成分能改善5×FAD小鼠的学习记忆障碍,减少Aβ过度沉积和老年斑形成,降低脑内炎症反应,改善APP/PS1转基因5×FAD小鼠脑内病理特征。

Objective:To study the effects of active ingredients of Tibetan medicine Wangla on the formation of senile plaques and inflammatory response in APP/PS1 double transgenic 5×FAD mice.Methods:A total of 20 four-month-old 5×FAD mice were randomly divided into model group(n=10)and active ingredients of Wangla group(n=10).Other 20 wild type(WT)mice of identical age and genetic background were randomly divided into normal control group(n=10)and active ingredients of Wangla control group(n=10).The normal control group and model group were given distilled water,and active ingredients of Wangla groups were given the corresponding drugs intragastrically.After 80 days of administration,the learning and memory function of each group was tested by Morris water maze test,the deposition of amyloid plaques in the brain was detected by immunohistochemical staining,the β-amyloid protein(Aβ)deposition and the expressions of ionic calcium binds the connector molecule 1(Iba1)and glial fibrillary acidic protein(GFAP)were detected by immunofluorescence staining and Western Blot.ELISA was used to detect the levels of inflammatory factors in brain.Results:Compared with the model group,the latency time of water maze of mice in active ingredients of Wangla group was significantly decreased,the times of crossing the target quadrant and residence time of mice were significantly increased,the deposition of brain senile plaques in brain,the expressions of Aβ,Iba1 and GFAP and the levels of TNF-α,IL-6,IL-β were significantly decreased(P<0.05 to P<0.001).Conclusion:The active ingredients of Wangla can improve learning and memory impairment of APP/PS1 transgenic 5×FAD mice,reduce Aβ deposition,senile plaques formation and the inflammatory response,improve pathological characteristics in the brain of APP/PS1 transgenic 5×FAD mice.