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目的:观察复方地黄对阿尔茨海默病(AD)模型大鼠学习记忆及Wnt通路相关蛋白表达的影响,探讨其作用机制。方法:采用Morris水迷宫筛选出健康Wistar大鼠,随机分为正常组,模型组,复方地黄高、低剂量(3.37,1.35 g·kg-1)组,加兰他敏(0.6 mg·kg-1)组。双侧海马区注射β-淀粉样蛋白(Aβ)23-35造模(每侧10μL),注射后第9天,ig相应药物,持续30d。采用水迷宫检测大鼠的学习记忆能力,用免疫组化、免疫印迹及实时荧光定量PCR检测大鼠海马Wnt通路相关蛋白轴蛋白(Axin)及细胞周期蛋白(Cyclin D1)表达。结果:模型组大鼠逃避潜伏期和首次到达原平台象限的时间均较正常组显著增多(P<0.01)。与模型组比较,加蓝他敏组及复方地黄高、低剂量组大鼠逃避潜伏期和首次到达原平台象限的时间明显减少(P<0.05)。复方地黄高剂量组较加兰他敏组首次到达原平台象限的时间减少(P<0.05)。免疫组化结果显示阳性表达Axin在胞质,Cyclin D1在胞核。免疫印迹法及实时荧光定量PCR结果显示,与正常组大鼠比较,模型组大鼠海马Axin mRNA及蛋白表达显著增高(P<0.01);与模型组比较,复方地黄高、低剂量组及加兰他敏组大鼠海马Axin mRNA及蛋白表达明显降低(P<0.05),Cyclin D1mRNA及蛋白表达明显升高(P<0.05)。结论:复方地黄可以改善AD模型大鼠学习记忆能力,其作用机制可能与降低AD大鼠海马Wnt通路相关蛋白Axin,增强Cyclin D1表达密切相关。
Objective: To observe the effects of compound Rehmanniae on learning and memory and Wnt pathway-related protein expression in Alzheimer’s disease (AD) model rats and to explore its mechanism. Methods: Healthy Wistar rats were screened by Morris water maze and randomly divided into three groups: normal group, model group, high and low doses of Dihuang (3.37,1.35 g · kg-1), galantamine 0.6 mg · kg- Group 1. Bilateral hippocampal areas were injected with β-amyloid (Aβ) 23-35 (10 μL per side) and on the 9th day after injection, the corresponding drug was administered for 30 days. The learning and memory abilities of rats were detected by water maze. The expression of Axin and Cyclin D1 in Wnt pathway of hippocampus was detected by immunohistochemistry, Western blotting and real-time fluorescence quantitative PCR. Results: The escape latency and the time to reach the original platform quadrant in model group were significantly higher than those in normal group (P <0.01). Compared with the model group, the escape latency and the time to reach the original platform quadrant in the high and low dose Rehmanniae group and the compound Rehmannia glutinosa group were significantly decreased (P <0.05). Compound Rehmannia high dose group than galanthamine group for the first time to reach the original platform quadrant time decreased (P <0.05). Immunohistochemistry showed Axin positive in the cytoplasm and Cyclin D1 in the nucleus. Immunoblotting and real-time fluorescent quantitative PCR results showed that Axin mRNA and protein expression in hippocampus of model group was significantly higher than that in normal group (P <0.01). Compared with model group, high and low dose of Dihuang compound The expression of Axin mRNA and protein in the hippocampus of the experimental group was significantly decreased (P <0.05), and the mRNA and protein expressions of Cyclin D1 were significantly increased (P <0.05). CONCLUSION: Compound Rehmanniae can improve the learning and memory abilities of AD model rats. The mechanism may be related to the reduction of Axin in Wnt pathway of hippocampus and the increase of Cyclin D1 expression in AD rats.