基于STAT3—GSK3β/PTEN/Akt通路探讨柴胡疏肝散抑制神经炎症治疗卒中后抑郁的机制

Post-stroke depression (PSD) seriously affects stroke rehabilitation and increases mortality. Microglia-mediated neuroinflammation is involved in the occurrence and development of PSD. Regulating the polarization of microglia is of great significance in inhibiting neuroinflammation. We previously found that Chaihu Shugan Powder was effective in the treatment of PSD by its role in the regulation.of inflammatory response. However, the key mechanism is unclear. In this project, firstly, the animal model would be proposed to confirm.the effect of Chaihu Shugan Powder on neuroinflammation. Then, a variety.of cell and molecular biology techniques are used in cell model to study the.effects of Chaihu Shugan Powder on microglia phenotype, inflammatory cytokines and pathways. Lastly, the molecular mechanism of regulating microglia.polarization is investigated by using RNA interference technology and inhibitors. This project is the first to elucidate the role of microglial polarization in the pathogenesis of PSD, to clarify the molecular mechanism of Chaihu Shugan Powder inhibiting neuroinflammation, and to provide a new direction and strategy for the treatment and research of PSD.

卒中后抑郁（PSD）严重影响卒中康复，并增加死亡率。小胶质细胞介导的神经炎症参与PSD的发生发展。调节小胶质细胞极化对抑制神经炎症有重要意义。我们前期发现柴胡疏肝散治疗PSD临床效果显著，其作用与调节炎症有关。但关键作用机制尚不明确。本项目拟通过动物模型验证柴胡疏肝散 抑制神经炎症的治疗作用；建立细胞模型，运用各种细胞分子生物学技术，研究柴胡疏肝散对小胶质细胞表型和其介导的神经炎症因子的影 响，探讨柴胡疏肝散调节炎症与小胶质细胞极化间的关系；并利用通路抑制剂和RNA干扰技术研究调控小胶质细胞极化的分子机制。本项目首次阐明小胶质细胞极化在PSD发病机制中的作用，明确柴胡疏肝散抑制神经炎症的分子机制，为PSD 治疗和研究提供新的方向和策略。

中风后抑郁被认为是卒中后最常见和最重要的神经精神后果。大约1/3的中风患者会经历严重的抑郁。柴胡疏肝散是治疗抑郁症的经典方剂。本项目旨在明确柴胡疏肝散对PSD大鼠模型的抗抑郁作用，探讨潜在的机制，并在体外氧葡萄糖剥夺(OGD)细胞模型中验证。本项目采用大脑中动脉闭塞(MCAO)联合慢性不可预测的轻度应激(CUMS)筛选抑郁大鼠。随后构建GSK3β腺病毒，并在体外模型成功过表达。进一步检测IL-1、IL-6、IL-10、TNF-α等炎症因子和Akt、p-Akt、CD16、CD206、PTEN、p PTEN、GSK3β、p- GSK3β、Stat3、p- Stat3等相关通路蛋白的表达，利用透射电镜观察突触超微结构。流式细胞仪显示，造模、GSK3β过表达病毒、柴胡疏肝散和JAK-STAT3抑制剂干预后，BV-2细胞由M1向M2型极化，表现为cd16 /32阳性细胞(M1细胞)被抑制，cd206阳性细胞(M2细胞)被促进。实验结果表明，柴胡疏肝散对PSD具有一定治疗作用。其作用机制可能是通过JAK/STAT3-GSK3β/PTEN/Akt通路调节小胶质细胞极化以抑制神经炎症。