Huang Sen, Zhan Zhaochun, Xu Fei, Liu Xiaolin, Fang Zhenning, Wu Wenbo, Liang Zhile, Liu Guoguo, Wang Mengyuan, da Silva Helena Soares, Luo Xin, Mo Kai
Department of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
Key Laboratory of Mental Health of the Ministry of Education, Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain-Inspired Intelligence, Guangdong-Hong Kong Joint Laboratory for Psychiatric Disorders, Guangdong Province Key Laboratory of Psychiatric Disorders, Guangdong Basic Research Center of Excellence for Integrated Traditional and Western Medicine for Qingzhi Diseases, Department of Neurobiology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.
Front Immunol. 2025 Apr 30;16:1569295. doi: 10.3389/fimmu.2025.1569295. eCollection 2025.
Rheumatoid arthritis (RA) is a chronic condition characterized by joint pain that significantly impairs patients' work and daily lives. The limited understanding of the pathological mechanisms underlying RA-related pain poses challenges for effective clinical pain management. spore powder (GLSP) has demonstrated therapeutic benefits in various diseases, with no reported toxicity or adverse effects.
This study investigates the role of neutrophils in the pathological mechanisms of RA-related pain using collagen-induced arthritis (CIA) mice and an neutrophil model. A combination of techniques, including animal models, flow cytometry, behavioral testing, cell adoptive transfer, and network pharmacology analysis, was employed to evaluate the effects and targets of GLSP on pain symptoms and neutrophil activity in CIA mice.
Flow cytometric analysis revealed the accumulation and activation of neutrophils in the paws of CIA mice. Furthermore, the levels of pro-inflammatory CD95 neutrophil subpopulations (N1 state) and ROS cells in the affected paws were positively correlated with the severity of mechanical allodynia and heat hyperalgesia observed in these mice. Our findings indicate that oral administration of GLSP significantly alleviates joint destruction, paw swelling, and pain hypersensitivity in CIA mice. Notably, GLSP reversed CIA-induced neutrophil accumulation, N1 polarization, and reactive oxygen species (ROS) production. Both network pharmacology target prediction and experimental validation indicated that GLSP inhibits N1 polarization and ROS production in neutrophils by modulating the TNF-α signaling pathway, thus exerting RA-specific analgesic effects.
In summary, this study offers new insights into the pathological mechanisms of RA-related pain and demonstrates that neutrophil accumulation, N1 polarization, and ROS production contribute to RA-related pain. GLSP alleviates RA-related pain by inhibiting the pro-inflammatory phenotype of neutrophils, highlighting its potential for clinical translation in the treatment of RA.
类风湿性关节炎（RA）是一种以关节疼痛为特征的慢性疾病，严重影响患者的工作和日常生活。对RA相关疼痛的病理机制了解有限，给有效的临床疼痛管理带来了挑战。灵芝孢子粉（GLSP）已在多种疾病中显示出治疗效果，且无毒性或不良反应报告。
本研究使用胶原诱导性关节炎（CIA）小鼠和中性粒细胞模型，研究中性粒细胞在RA相关疼痛病理机制中的作用。采用包括动物模型、流式细胞术、行为测试、细胞过继转移和网络药理学分析等多种技术，评估GLSP对CIA小鼠疼痛症状和中性粒细胞活性的影响及作用靶点。
流式细胞术分析显示CIA小鼠爪中中性粒细胞的积聚和活化。此外，受累爪中促炎性CD95中性粒细胞亚群（N1状态）和ROS细胞的水平与这些小鼠中观察到的机械性异常性疼痛和热痛觉过敏的严重程度呈正相关。我们的研究结果表明，口服GLSP可显著减轻CIA小鼠的关节破坏、爪肿胀和疼痛超敏反应。值得注意的是，GLSP逆转了CIA诱导的中性粒细胞积聚、N1极化和活性氧（ROS）产生。网络药理学靶点预测和实验验证均表明，GLSP通过调节TNF-α信号通路抑制中性粒细胞中的N1极化和ROS产生，从而发挥RA特异性镇痛作用。
总之，本研究为RA相关疼痛的病理机制提供了新的见解，并表明中性粒细胞积聚、N1极化和ROS产生与RA相关疼痛有关。GLSP通过抑制中性粒细胞的促炎表型减轻RA相关疼痛，突出了其在RA治疗中临床转化的潜力。
