Zhang Yi, Luo Xiao-Ru, Zhou Yu-Hang, Xue Qian-Yin, Wu Shu-Ya, Xu Jing-Yu, Li Guo-Hui, Liu Bu-Ping, Zhang Ming-Jia, Tong Guang-Dong, Ao Hai-Qing, Liao Mian-Mian
The Affiliated Dongguan Songshan Lake Central Hospital, Guangdong Medical University, Dongguan, 523326, China.
Guangzhou University of Chinese Medicine, Guangzhou, 511400, China.
J Ethnopharmacol. 2025 Aug 29;352:120184. doi: 10.1016/j.jep.2025.120184. Epub 2025 Jun 21.
Xiaoyaosan (XYS), a classical herbal formulation indicated for "depression syndrome", can improve chronic stress-related mood disorders and immune disorders through multi-target modulation of inflammatory mediators, although its precise mechanism in regulating Th17/Treg balance remains to be clarified.
This study aims to elucidate the regulatory impact of XYS on the peripheral Th17/Treg immunological imbalance generated by chronic psychological stress through modulation of the FKBP5/NF-κB signaling pathway, thereby establishing a mechanistic foundation for developing innovative interventions with dual stress-adaptogenic and immunoregulatory properties.
FKBP5 knockout (FKBP5) and C57BL/6 mice underwent a 35-day chronic unpredictable mild stress (CUMS) paradigm. C57BL/6 mice were subsequently treated with XYS (1.61, 3.22, and 6.44 g/kg/d), fluoxetine (3 mg/kg/d). Behavioral phenotyping included the Open Field Test (OFT), Tail Suspension Test (TST), Forced Swimming Test (FST) and Sucrose preference test (SPT). Serum inflammatory cytokines (IL-17A, IL-23, IL-10, TNF-α) measured by enzyme-linked immunosorbent test (ELISA). Splenic Th17/Treg subset ratios were analyzed using multicolor flow cytometry. Western blotting evaluated FKBP5 expression and NF-κB pathway dynamics (IκB-α degradation, P65 nuclear translocation). UPLC-HRMS-based phytochemical profiling coupled with molecular docking elucidated XYS-FKBP5 interactions. FKBP5/IKKα/β complex formation was validated through confocal immunofluorescence (IF) colocalization and reciprocal co-immunoprecipitation (CO-IP) assays.
CUMS mice exhibited significant depression-like behaviors and hippocampal neuronal damage, concomitant with elevated serum corticosterone levels and Th17/Treg immune dysregulation. Medium- and high-dose XYS interventions substantially ameliorated these behavioral abnormalities, reversed hippocampal pathological alterations, suppressed pro-inflammatory responses, and rectified Th17/Treg imbalance. Crucially, XYS exerted therapeutic effects through targeted suppression of FKBP5-IKK complex interaction, effectively blocking NF-κB nuclear translocation, thereby restoring Th17/Treg homeostasis.
XYS ameliorates chronic psychological stress-induced Th17/Treg immune imbalance through FKBP5-targeted suppression of NF-κB hyperactivation. This multicomponent synergistic mechanism establishes a novel therapeutic strategy for stress-associated inflammatory comorbidities, particularly depression with immune dysregulation.
逍遥散（XYS）是一种用于治疗“郁证”的经典中药方剂，可通过多靶点调节炎症介质来改善慢性应激相关的情绪障碍和免疫紊乱，但其调节Th17/Treg平衡的确切机制仍有待阐明。
本研究旨在阐明逍遥散通过调节FKBP5/NF-κB信号通路对慢性心理应激所致外周Th17/Treg免疫失衡的调节作用，从而为开发具有双重应激适应原性和免疫调节特性的创新干预措施奠定机制基础。
FKBP5基因敲除（FKBP5）小鼠和C57BL/6小鼠接受为期35天的慢性不可预测温和应激（CUMS）模型。随后，C57BL/6小鼠分别接受逍遥散（1.61、3.22和6.44 g/kg/d）、氟西汀（3 mg/kg/d）治疗。行为表型分析包括旷场试验（OFT）、悬尾试验（TST）、强迫游泳试验（FST）和蔗糖偏好试验（SPT）。采用酶联免疫吸附试验（ELISA）检测血清炎症细胞因子（IL-17A、IL-23、IL-10、TNF-α）。使用多色流式细胞术分析脾脏Th17/Treg亚群比例。蛋白质免疫印迹法评估FKBP5表达和NF-κB信号通路动态变化（IκB-α降解、P65核转位）。基于超高效液相色谱-高分辨质谱的植物化学图谱分析结合分子对接阐明逍遥散与FKBP5的相互作用。通过共聚焦免疫荧光（IF）共定位和相互免疫共沉淀（CO-IP）试验验证FKBP5/IKKα/β复合物的形成。
CUMS小鼠表现出明显的抑郁样行为和海马神经元损伤，同时血清皮质酮水平升高和Th17/Treg免疫失调。中、高剂量逍遥散干预可显著改善这些行为异常，逆转海马病理改变，抑制促炎反应，并纠正Th17/Treg失衡。至关重要的是，逍遥散通过靶向抑制FKBP5-IKK复合物相互作用发挥治疗作用，有效阻断NF-κB核转位，从而恢复Th17/Treg稳态。
逍遥散通过靶向抑制FKBP5来抑制NF-κB过度激活，从而改善慢性心理应激诱导的Th17/Treg免疫失衡。这种多成分协同机制为应激相关炎症合并症，特别是伴有免疫失调的抑郁症建立了一种新的治疗策略。
