基于行为和网络药理学的蒙古医学Zadi-5在抑郁症大鼠模型中的分析
Summary
本方案描述了一种对蒙古传统药物Zadi-5在抑郁症中的治疗效果的行为测试验证和生物信息学预测的方法。
Abstract
Zadi-5是一种传统的蒙古药物，广泛用于治疗抑郁症和刺激症状。尽管Zadi-5对抑郁症的治疗作用已在先前报道的临床研究中得到证实，但药物中存在的活性药物化合物的身份和影响尚未完全阐明。本研究使用网络药理学来预测药物成分并鉴定Zadi-5药丸中的治疗活性化合物。在这里，我们建立了慢性不可预测的轻度应激（CUMS）大鼠模型，并进行了开放场试验（OFT）、莫里斯水迷宫（MWM）分析和蔗糖消耗试验（SCT），以研究Zadi-5在抑郁症中的潜在治疗效果。本研究旨在证明 Zadi-5 对抑郁症的治疗效果，并预测 Zadi-5 对抗该疾病的关键途径。氟西汀组（阳性对照）和Zadi-5组的纵横评分（OFT ）、SCT和跨区数显著高于未处理的CUMS组大鼠（P < 0.05）。根据网络药理学分析结果，发现PI3K-AKT通路对Zadi-5的抗抑郁作用至关重要。
Introduction
抑郁症，也称为重度抑郁症 （MDD），是一种严重的神经精神疾病，导致社会医疗和经济负担日益加重。由于相关的复杂性、发病率和死亡率，已经进行了大量研究以寻找该疾病的补救措施 1,2。根据世界卫生组织的一项心理健康调查，目前全球约有 3.5 亿人患有抑郁症及其相关症状。据预测，到 2030 年，抑郁症将超过癌症和心血管疾病，成为全球疾病负担的主要原因。因此，在不久的将来，抑郁症的预防和治疗将成为全球优先事项3.MDD的发病机制尚未阐明。尽管如此，它通常归因于以下因素：遗传易感性、下丘脑-垂体-肾上腺轴功能障碍、神经递质分泌减少、神经免疫失调诱导的神经炎症、细胞凋亡和细胞增殖减少 4,5。
在这些因素中，神经免疫失调诱导的神经炎症和神经营养因子分泌改变因其在抑郁症和许多其他精神疾病发展中的作用而受到特别关注6。在过去的十年中，学者们已经证明海马体是再生神经功能的主要场所，并参与调节情绪和认知。在这方面，海马神经元被认为是正在开发的抗抑郁药物的新治疗靶点 7,8。此外，据报道，海马体还参与学习和巩固记忆的短期和长期记忆。具体来说，海马CA1区域锥体神经元的缺乏会导致逆行和顺行性遗忘症9。典型的抗抑郁治疗策略旨在增强海马齿状回的细胞增殖和神经发生。基于药物化学技术合成的天然产物衍生化合物和小分子被认为是治疗各种神经精神疾病的创新治疗剂的主要来源。
蒙古族传统药物历史悠久，理论医学体系得到充分支持，是蒙古高原游牧民族的后裔，由于各种药用成分协同作用，产生协同作用，这些药物表现出多靶点和多通路效应。Zadi-5 是此类药物中公认的配方，最早记录在“高石博士的临床经验”中，由一位杰出的蒙古临床医生高石博士（1804-1876 年）撰写。在蒙古，使用这些药丸治疗痛苦、心悸、刺激和心脏刺痛的症状已经是很长一段时间的临床实践10,11。此外，Zadi-5 已被证明可缓解受影响患者的中风后抑郁症12。最近对 CUMS 的实验研究表明，Zadi-5 制剂通过调节中枢神经递质13 来缓解抑郁症;事实上，在 Zadi-5 中，已经检测到脑源性神经营养因子 （BDNF） 和酪氨酸激酶受体 B （TrkB） 水平升高，并与抑郁症大鼠模型中的学习和记忆改善相关14。然而，Zadi-5缓解抑郁症的确切作用机制尚未阐明。
本研究旨在通过行为测试证明Zadi-5对大鼠抑郁症的治疗效果，并使用中医药系统药理学（TCMSP）和瑞士靶点预测法鉴定Zadi-5的成分，以预测蒙古传统药物Zadi-5治疗抑郁症的潜在机制。
Protocol
所有实验方案均经内蒙古医科大学动物实验伦理护理委员会批准，并遵循美国国立卫生研究院动物护理伦理指南。将8周龄（200g±20g）的雄性Sprague Dawley（SD）大鼠饲养在受控温度（22°C±2°C）和湿度（55%±15%）的房间中，在12小时/12小时调节的光/暗循环下，持续1周。网络药理学分析的工作流程见 图1 。 1.大鼠行为测试 建立CUMS大鼠模型…
Representative Results
动物行为测试CUMS诱导的大鼠抑郁模型中的行为测试结果在CUMS刺激前，测试组之间的OFT评分、蔗糖消耗量和MWM分析没有显著差异。建立CUMS模型后，MOD组的纵向和水平得分均低于CON组（P < 0.05）。与MOD组相比，POS组和Zadi-5组的纵向和水平得分显著更高（P < 0.05）（图2A，B）。 第0天，试验组的蔗糖消耗量（%…
Discussion
抑郁症是一种以情绪低落、快感缺乏和精力不足为特征的精神疾病。这种疾病伴有分心、认知功能障碍、社交退缩、失眠、性功能障碍和胃肠道疾病18,19。在抑郁症的研究中，建立动物模型对于理解新药的病理机制和作用至关重要。在这项研究中，通过协议第 2 节和表 1 中描述的刺激建立了 CUMS 诱导的大鼠抑郁模型，以模拟来自社会、家庭?…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
我们感谢中国内蒙古医科大学蒙古医学院提供的仪器和实验室。本研究得到了国家自然科学基金（81760762）和内蒙古卫生健康委员会科技计划项目（202201300）的支持。
Materials
| Cytoscape software | version 3.7.0 | ||
| Fluoxetine | Lilly Suzhou Pharmaceutical Co., Ltd | J20160029 | |
| Morris water maze video trail analysing system | Tai Meng Tech Co., Ltd | WMT-200 | |
| Sprague Dawley rats | Beijing Biotechnology Co., Ltd, China | SCXK (JING) 2016-0002 | |
| video tracking system | Tai Meng Tech Co., Ltd | ZH-ZFT | |
| Zadi-5 pill | Pharmaceutical Preparation Center of International Mongolian Hospital, Inner Mongolia, China | M1301006 |
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Citazione di questo articolo
Wu, R., Zu, W., Wu, L., Su, S., Su, N., Qi, L., Wu, R., Sun, W., Hu, R. Behavioral and Network Pharmacology-Based Analyses for the Traditional Mongolian Medicine Zadi-5 in a Rat Model of Depression. J. Vis. Exp. (192), e64832, doi:10.3791/64832 (2023).