苦参子提取物及其活性成分苦参子二肤的杭抑郁作用研究
文献类型：学位论文
| 作者 | 朱天 |
| 答辩日期 | 2023-06 |
| 文献子类 | 硕士 |
| 授予单位 | 中国科学院大学 |
| 授予地点 | 中国科学院心理研究所 |
| 其他责任者 | 郭建友 |
| 关键词 | 苦参子提取物 抑郁 Wnt/β-catenin信号转导通路 行为学实验 苦参子二肤 |
| 学位名称 | 理学硕士 |
| 学位专业 | 发展与教育心理学 |
| 其他题名 | Study on the antidepressant effect of Sophora flavescens seed extract and its active ingredient Sophora flavescens seed dipeptide |
| 中文摘要 | Depression is a common mental disorder, and its treatment currently mainly includes medication, psychotherapy, and electroconvulsive therapy. Antidepressants, including SSRIs, SNRIs, and tricyclics, can regulate neurotransmitters and relieve depression symptoms, However, the current clinical antidepressant chemistry is generally prone to drug resistance, serious adverse effects and relapse after discontinuation of medication. Therefore, there is an urgent need to find better treatment methods and drugs for depression. Multiple chemical components in Chinese medicine synergistically promote each other, which can effectively avoid the side effects of the medicinal effect of a single component. Sophora flavescens is a traditional Chinese medicine that is believed to have antidepressant effects by regulating neurotransmitter levels and improving the function of the nervous system. In addition, Sophora flavescens seed extract can alleviate depression symptoms caused by chronic stress and increase the number of neurons. There is a certain relationship between depression and neuroinflammation. Neuroinflammation is a type of inflammatory response that mainly involves the inflammatory response of the nervous system. Studies have shown that neuroinflammation may be one of the causes of depression. During the neuroinflammation process, inflammatory cells release a series of inflammatory factors, including tumor necrosis factor-α, interleukin-1β，and interleukin-6. These factors can affect the activity of neurons and the level of neurotransmitters. Therefore, exploring the relationship between depression and neuroinflammation may help to better understand the pathogenesis of depression and provide new ideas for the treatment of depression.Some studies have shown that the Wnt/β-catenin pathway may play a certain role in neuroinflammation. In addition, the expression level of beitβ-catenin in neurons may also be affected by neuroinflammation. Recent studies have shown that abnormalities in the Wnt/β-catenin pathway are closely associated with depression, schizophrenia and other psychiatric disorders (Zhou Zongpu, 2018), and that antidepressant drugs can exert a better antidepressant effect by altering the expression of Wnt, GSK-3β，p-catenin and CyclinD 1 proteins that are abnormally altered in brain tissue (Wang Rui, 2017). Therefore, the Wnt/β-catenin pathway may be an important area in the study of neuroinflammation and future studies will help to gain insight into the role of this pathway in neuroinflammation. Objective: The purpose of this study was to investigate the antidepressant effect and mechanism of sophora flavescens seed dipeptide, a unique active ingredient of Sophora flavescens seed extract. The study used intraperitoneal injection of lipopolysaccharide (LPS) to establish a depression model in mice, and conducted open field test, elevated plus maze test, and forced swimming test to evaluate the antidepressant efficacy of Sophora flavescens extract;Western blot method was used to detect the protein expression levels of G1/S-specific cycle protein-D1 (Cyclin D1), Wntl protein,p-catenin, and glycogen synthase kinase-3β (p-GSK-3β)in the mouse brain tissue; The TUNEL method was used to detect the apoptosis of hippocampal cells; Isolation of mouse brain tissue for the determination of inflammatory factors IL-1，IL-6, TNF-a and anti-inflammatory factor IL-10 in brain tissue; Autodock Vina and Pymol software were used to dock and verify the mechanism of sophora flavescens seed dipeptide and its key targets, confirming the antidepressant effect of Sophora. flavescens seed dipeptide. Method: The results showed that Sophora flavescens seed extract had a significant impact on the depressive-like behavior of mice. Compared with the control group, the total distance traveled, movement speed, and distance traveled in the center zone in the open field test, as well as the time spent in the open arms of the elevated plus maze test were significantly decreased, and the immobility time in the forced swimming test was significantly increased in the model group. However, compared with the model group, the middle and high dose groups of Sophora. flavescens seed extract showed increased movement speed and distance traveled in the center zone in the open field test, increased time spent in the open arms of the elevated plus maze test, and decreased immobility time in the forced swimming test; Western blot results showed that the protein expression levels of Wntl andp-catenin were significantly decreased, while Cyclin D1 and p-GSK-3β were significantly increased after LPS injection in mice. Compared with the model group, the middle and high dose groups of Sophora flavescens seed extract showed a significant increase in the protein expression of Wntl andp一catenin, and a significant decrease in Cyclin D1 and p-GSK-3β;TUNEL staining showed that the apoptosis rate of hippocampal cells in the model group was significantly increased compared with the normal group, and the middle and high dose groups of Sophora flavescens seed extract showed a significant decrease in the apoptosis rate of hippocampal cells; Compared with the control group, the levels of inflammatory factors TNF-α, IL-6 and IL-1β were significantly increased and the levels of anti-inflammatory factor IL-10 were significantly decreased in the model group of mice; Compared with the model group, the levels of inflammatory factors TNF-α, IL-6 and IL-1β were decreased in the mid-dose and high-dose groups of bitter ginseng seeds, and the levels of anti-inflammatory factor IL-10 were increased in the mid-dose and high-dose groups of bitter ginseng seeds; The binding energy produced by the combination of ligand molecule sophora flavescens seed dipeptide with p-GSK-3β and Cyclin D 1 protein indicated that the ligand molecule sophora flavescens seed dipeptide was stably bound to them; After intervention with sophora flavescens seed dipeptide, the middle and high dose groups of mice showed increased movement speed and distance traveled in the center zone in the open field test, increased time spent in the open arms of the elevated plus maze test, and decreased immobility time in the forced swimming test. Result: Therefore, Sophora flavescens seed extract can effectively improve the severity of depression in LPS-induced depressed mice, and its molecular mechanism is related to the inhibition of neuroinflammation mediated by the Wnt/β-catenin signaling pathway. The main active ingredient of the extract with antidepressant activity is possibly Sophora flavescens seed dipeptide. This provides a new theoretical basis for research on depression and strengthens research on Sophora flavescens, which can lead to its resource utilization and industrial development. This will have a positive promoting effect on the comprehensive utilization and industrial economic development of Sophora plant resources and better serve the public. |
| 英文摘要 | 抑郁症是一种常见精神疾病，治疗包括:药物治疗、心理治疗及电疗。抗抑郁药主要SSRI类、SNRI类、三环类等药物，这些药物能调节神经递质，缓解抑郁症状，但目前临床抗抑郁化学药普遍存在易产生耐药性、不良反应严重、停药后易复发等问题。因此，迫切需要寻找更好的抑郁症治疗方法及药物。中药多种化学成分相互协同促进，可有效避免单一成分药效副作用。传统中药材苦参子，被认为具有抗抑郁功效，可通过调节神经递质水平，改善神经系统功能，发挥抗抑郁作用。此外，苦参子提取物能减轻慢性压力导致的抑郁症状，并增加神经元数量。抑郁症和神经炎症间存在一定关联。神经炎症是一种炎症反应类型，涉及神经系统炎症反应。研究表明，神经炎症可能导致抑郁症发生和持续原因之一。神经炎症过程中，炎症细胞释放一系列炎症因子，包括肿瘤坏死因子-α、白细胞介素-1β和白细胞介素-6等。这些因子可影响神经元活动和神经递质水平。因此，探究抑郁症和神经炎症关系，可能有助于更好理解抑郁症发病机制，同时也为抑郁症治疗提供新思路。研究表明，Wnt/β-catenin通路可能作用于神经炎症中。Wnt/(β-catenin通路是一种重要信号转导通路，它参与调节多种生物学过程。近年研究表明，Wnt/p-catenin通路异常与抑郁症、精神分裂症等精神疾病密切相关(周宗朴，2018)，抗抑郁药物通过改变脑组织异常变化Wnt, GSK-3β、β-catenin及CyclinD 1蛋白表达(王睿，2017 )，发挥较好的抗抑郁作用。因此，Wnt/β-catenin通路可能是神经炎症研究重要领域，未来研究将有助于深入了解这种通路神经炎症作用。 目的:为了进一步探讨苦参子提取物抗抑郁作用及其机制，并其特有活性成分苦参子二肤抗抑郁作用。 方法:本课题应用腹腔注射脂多糖(Lipopolysaccharide，LPS)建立小鼠抑郁模型，利用旷场实验、高架十字迷宫实验和强迫游泳实验考察苦参子提取物抗抑郁药效;采用蛋白免疫印迹法(Western blot方法)检测小鼠脑组织中G1/S-特异性周期蛋白-D1 （Cyclin D1)、Wntl蛋白、β-连环蛋白(β-catenin)、糖原合成酶激酶3β(p-GSK-3β）的蛋白表达水平;通过原位末端标记法(TUNEL法) 检测海马细胞凋亡情况;分离小鼠脑组织，检测脑组织中炎症因子IL-1, IL-6, TNF-α和抗炎症因子IL-10含量;利用Autodock Vina脚本将小分子苦参子二肤与关键靶点CyclinD 1和p-GSK-3β蛋白进行对接;利用Pymol软件对配体一受体蛋白复合物相互作用的分析，验证并初步确认苦参子二肤抗抑郁作用。 结果表明苦参子提取物影响小鼠抑郁样行为:与对照组相比，模型组小鼠旷场总运动距离，旷场运动速度和中间区运动距离，高架十字迷宫开臂停留时间均显著降低，强迫游泳实验不动时间显著增加，与模型组相比，苦参子中剂量组及苦参子高剂量组小鼠旷场实验旷场运动速度增加，中间区运动距离增加，高架十字迷宫开臂停留时间提升，强迫游泳实验不动时间降低;Western blot结果显示，小鼠LPS注射后Wnt1及β-catenin蛋白表达水平显著降低，Cyclin D 1及p-GSK-3β蛋白表达水平显著增加，与模型组相比，苦参子中剂量组和高剂量组小鼠Wntl 蛋白和β-catenin蛋白表达显著增加，而Cyclin D 1蛋白和p-GSK-3β蛋白表达显著降低;TUNEL染色法显示，模型组小鼠海马细胞凋亡率较正常组显著增加，与模型组相比，苦参子中剂量和高剂量组小鼠海马细胞凋亡率显著降低;与对照组相比，模型组小鼠炎性因子TNF-α, IL-6和IL-1β的含量显著上升，抗炎性因子IL-10的含量显著下降，与模型组相比，苦参子中剂量组和高剂量组炎性因子TNF-α, IL-6和IL-1β的含量下降，苦参子中剂量组和高剂量组的抗炎性因子IL-10的含量有所增加;苦参子二肤与p-GSK-3β蛋白和Cyclin D 1蛋白所产生的结合能表明了配体分子苦参子二肤稳定地与其结合;苦参子二肤干预LPS注射小鼠后，中剂量组及高剂量组小鼠旷场运动速度增加，旷场中间区运动距离增加，高架十字迷宫开臂停留时间增加，强迫游泳实验不动时间降低。 结论:因此苦参子提取物可有效改善LPS所致抑郁小鼠的抑郁严重程度，其分子机理与抑制Wnt/β-catenin信号转导通路介导的神经炎症有关，该提取物抗抑郁主要活性成分可能是苦参子二肤，为抑郁症相关研究提供新的依据，加强苦参子研究，进而资源化利用和产业化开发，将对苦参植物资源综合利用和产业经济发展起到积极推动作用，更好服务于大众。 |
| 语种 | 中文 |
| 源URL | [http://ir.psych.ac.cn/handle/311026/46667]  |
| 专题 | 心理研究所_健康与遗传心理学研究室 |
| 推荐引用方式 GB/T 7714 | 朱天. 苦参子提取物及其活性成分苦参子二肤的杭抑郁作用研究[D]. 中国科学院心理研究所. 中国科学院大学. 2023. |
入库方式： OAI收割
来源：心理研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。