基于Nrf2/ARE信号通路的广西黑蚁抗抑郁功效物质基础及作用机制研究

Depression is characterized by long term depressing psychotic symptoms, with high incidence, high recurrence,and high rate of suicide, which will become the second major disease threatening public health. However, there is still no safe and effective antidepressant as yet. Depression belongs to the Yu syndrome in Traditional Chinese Medicine(TCM).Kidney deficiency and stagnation of liver qi are its major pathogenesis. Tonifying kidney and regulating qi activity achieved good results in TCM clinical practice. Polyrhachis vicina Roger(PVR) is the trueborn medicine of Guangxi, which was known as its effect on tonifying kidney energy, clearing and activating the channels and collaterals. Our previous studies showed that PVR extract could improve the SOD level of rodent animal significantly, attenuate hypothermia and weakened induced by reserpine, suggested that it has potential for the treatment of depression. On this basis, the present study would prepare the different active site of PVRS by preparative liquid chromatography extraction method. Behavioral tests, molecular biology, immunohistochemistry and immunofluorescence techniques would be employed to investigate the characters of anti-depressive effect induced by PVR exctract. It's mechanism would be revealed by observing its effect on the dynamics of Nrf2 signaling pathway, which is the most relevant signaling pathways to oxidative stress. This study would provide a solid theoretical basis for the developed of antidepressant with clear structure and mechanism from Guangxi.

抑郁症是一种以持续的心境低落为主要表现的精神症状，发病率高、是诱发自杀率最高的疾病，即将成为威胁人类健康第二大疾病，但目前尚无安全有效的治疗药物。抑郁症在中医属于郁证范畴，肾虚肝郁为其主要病机。临床以益肾调气、解郁安神为法则，取得了良好的治疗效果。黑蚁（拟黑多刺蚁）为广西道地药材，具有补肾益精，通经活络之功效。我们前期研究表明广西黑蚁提取物可显著提升啮齿类动物SOD水平，缓解由于利血平引发的体温降低、活动力减弱等现象，提示其具有抗抑郁的潜能。在此基础上，本项目拟采用制备液相色谱提取方法制备广西黑蚁不同活性部位，采用行为学、分子生物学、免疫组织化学、免疫荧光等技术，考察广西黑蚁不同活性部位抗抑郁功效特点，并试图通过与氧化应激密切相关的Nrf2信号通路动力学变化阐明广西黑蚁抗抑郁作用机制，为从广西特色药中研制出物质结构清晰、作用机制明确的抗抑郁症药物奠定坚实的理论基础。

本项目通过构建小鼠抑郁模型，给予广西特产黑蚁活性组分（AFPR）治疗，检测小鼠悬尾试验(TST)、强迫游泳试验(FST)和野外试验(OPT)，测定AFPR对利血平诱导的上睑下垂、低温和失语症的影响，单胺及其代谢产物的水平，以及海马和前额叶皮层单胺氧化酶(MAO)的活性，阐明了AFPR具有抗抑郁作用，通过调节脑内单胺类神经递质系统治疗抑郁症的作用机制。通过构建慢性不可预见性温和刺激大鼠体内外抑郁模型，通过大鼠糖水偏好、强迫游泳等行为学实验以及细胞免疫荧光、qRT-PCR、western blot等实验，检测NF-κB炎症信号通路的表达，进一步阐明AFPR抗抑郁作用的多靶点性。通过AFPR处理皮质酮损伤的星形胶质细胞进行lncRNA-seq测序并通过qRT-PCR等实验，初步阐明AFPR可能通过lncRNA-GAS5和lncRNA-SNHG1调控Cx43及其介导的GJIC功能而发挥抗抑郁作用。在此基础上，本项目还开展拓展实验，探究AFPR对肿瘤、瘙痒和系统性红斑狼疮等疾病的治疗作用及相关机制。本项目的研究内容已全部完成，本研究为研制具有自主知识产权的抗抑郁药物奠定基础。.在本项目的支持下，发表论文 6 篇，其中 SCI 2 篇，中文核心期刊 4 篇；获得国家发明专利 1 项。