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目的:观察补肾化痰益智方(BSHT)对APPV717I转基因小鼠干预效果,揭示其可能作用机制。方法:将3月龄APPV717I转基因小鼠120只分为2批,每批60只,每批又随机分为5组,分别为多奈哌齐(0.000 92 g·kg~(-1)·d~(-1))组,补肾化痰益智方低、中、高剂量组(5.2,10.4,20.8 g·kg~(-1)·d~(-1)),模型组(APP),每组12只(雌雄各半);同月龄遗传背景相同的C57BL/6J小鼠24只作为正常组,灌胃给药,模型组及正常组予等容积双蒸水灌胃,每日1次。第1批小鼠给药8个月,第2批给药4个月,最后同时以Morris水迷宫进行行为学检测,分别以酶联免疫吸附测定(ELISA)和免疫印迹法测定(Western blot)海马神经元Tau蛋白磷酸化(p-Ser 199/202,Tau-1)的表达水平。结果:行为学实验结果表明,与正常组比较,模型组小鼠的潜伏期有所延长(P<0.05,P<0.01);各治疗组与模型组比潜伏期明显缩短(P<0.05)。ELISA和Western blot实验结果表明,与正常组比较,模型组小鼠海马神经元的Ser199/202磷酸化位点磷酸化程度显著升高(P<0.01);各治疗组Ser199/202磷酸化位点磷酸化水平较模型组比明显缩短(P<0.05,P<0.01)。结论:补肾化痰益智方可以改善Ser199/202磷酸化位点磷酸化程度,防止神经原纤维缠结(NFT)的形成,从而改善APPV717I转基因小鼠早期的学习记忆能力。
Objective: To observe the intervention effect of Bushen Huatan Yizhi Recipe (BSHT) on APPV717I transgenic mice, and to reveal its possible mechanism. Methods: 120 APPV717I transgenic mice of 3 months old were divided into two groups of 60, each group was randomly divided into 5 groups: donepezil (0.000 92 g · kg -1 · d ~ (- 1) group, Bushen Huatan Yizhi Fang low, medium and high dose groups (5.2,10.4,20.8 g · kg -1 · d -1), model group (APP) 24 male C57BL / 6J mice with the same genetic background on the same month as the normal group, intragastric administration, the model group and the normal group were intragastrically given the same volume of double distilled water once a day. The first batch of mice was administered for 8 months and the second batch for 4 months. At the same time, behavioral tests were performed in the Morris water maze, and were determined by ELISA and Western blot, respectively Expression of Tau phosphorylation (p-Ser 199/202, Tau-1) in hippocampal neurons. Results: The results of behavioral experiments showed that compared with the normal group, the incubation period in the model group was prolonged (P <0.05, P <0.01). The latency in each treatment group and model group was significantly shorter than that in the normal group (P <0.05). The results of ELISA and Western blot showed that the phosphorylation of Ser199 / 202 phosphorylation in hippocampal neurons of model group was significantly higher than that of normal group (P <0.01). Ser199 / 202 phosphorylation sites Compared with the model group, the phosphorylation level was significantly shorter (P <0.05, P <0.01). Conclusion: Bushen Huatan Yizhi Decoction can improve the phosphorylation of Ser199 / 202 phosphorylation site and prevent the formation of neurofibrillary tangles (NFT), so as to improve the early learning and memory ability of APPV717I transgenic mice.