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目的观察银杏叶提取物(EGB)金纳多对血管性痴呆(VD)小鼠海马结构神经元型一氧化氮合酶(nNOS)表达的影响,探讨银杏叶提取物对血管性痴呆的治疗作用。方法复制VD小鼠模型,利用Y-迷宫检测VD模型小鼠学习记忆能力及不同剂量EGB治疗组的改善作用,实时定量PCR方法检测不同剂量EGB对VD小鼠海马结构中NOSmRNA转录水平的影响,组织化学和免疫组化方法研究其对NOS和nNOS蛋白表达的影响。结果行为学结果显示VD模型组和各治疗组小鼠均比对照组小鼠Y-迷宫学习记忆训练次数明显增多(<0.05),高低两个剂量EGB治疗组迷宫学会次数与模型组相比明显减少(<0.05),有剂量依赖性。实时定量PCR结果显示在背侧海马VD模型组nNOS mRNA转录水平显著提高,而高低EGB治疗组的nNOS mRNA转录水平显著降低(<0.05)。组织化学和免疫组化结果显示在海马结构CA1区VD模型组比对照组NOS和nNOS阳性神经元的数量明显增多(<0.05),高低两个EGB治疗组与VD模型组相比阳性神经元数量明显减少(<0.05)。结论银杏叶提取物对VD小鼠神经元有保护作用,可能与减少海马结构NOS的表达相关。
Objective To observe the effect of Ginkgo biloba extract (GGB) on the expression of nitric oxide synthase (nNOS) in the hippocampal formation of vascular dementia (VD) mice and to explore the therapeutic effect of Ginkgo biloba extract on vascular dementia . Methods VD mouse model was duplicated, the learning and memory abilities of VD model mice were examined by Y-maze, and the effect of different doses of EGB treatment group was evaluated. The effect of different doses of EGB on NOS mRNA transcription in hippocampal formation of VD mice was detected by real- Histochemical and immunohistochemical methods to study its NOS and nNOS protein expression. Results The results of behavioral study showed that the number of Y-maze learning and memory training in VD model group and each treatment group was significantly higher than that in control group (<0.05), and the number of labyrinthine learning and memory in both high and low EGB treatment groups was significantly higher than that in model group Decreased (<0.05), and dose-dependent. Real-time quantitative PCR results showed that the nNOS mRNA transcription level in VD model group was significantly increased, while the level of nNOS mRNA transcription in high and low EGB treatment group was significantly lower (P <0.05). Histochemistry and immunohistochemistry results showed that the number of NOS and nNOS positive neurons in the hippocampal CA1 region VD model group was significantly higher than that in the control group (<0.05), the number of positive neurons in the two EGB treatment groups was higher than that in the VD model group Significant reduction (<0.05). Conclusion Ginkgo biloba extract has a protective effect on neurons in VD mice, which may be related to the decrease of NOS expression in hippocampal formation.