State Key Laboratory of Military Stomatology, National Clinical Research Center for Oral Diseases, Shaanxi International Joint Research Center for Oral Diseases, Department of Pharmacy, School of Stomatology, Fourth Military Medical University, Xi'an, China.
Department of Pharmacology, School of Pharmacy, Fourth Military Medical University, Xi'an, China.
Phytother Res. 2024 Nov;38(11):5417-5440. doi: 10.1002/ptr.8325. Epub 2024 Sep 12.
Long-term inflammation can cause chronic pain and trigger patients' anxiety by sensitizing the central nervous system. However, effective drugs with few side effects for treating chronic pain-induced anxiety are still lacking. The anxiolytic and anti-inflammatory effects of ruscogenin (RUS), an important active compound in Ophiopogon japonicus, were evaluated in a mouse model of chronic inflammatory pain and N9 cells. RUS (5, 10, or 20 mg/kg/day, i.g.) was administered once daily for 7 days after CFA injection; pain- and anxiety-like behaviors were assessed in mice. Anti-inflammatory effect of RUS (0.1, 1, 10 μM) on N9 microglia after LPS treatment was evaluated. Inflammatory markers (TNF-α, IL-1β, IL-6, CD86, IL-4, ARG-1, and CD206) were measured using qPCR. The levels of IBA1, ROS, NF-κB, TLR4, P-IKK, P-IκBα, and P65, MAPKs (ERK, JNK, and P38), NLRP3 (caspase-1, ASC, and NLRP3) were detected by Western blotting or immunofluorescence staining. The potential target of RUS was validated by molecular docking and adeno-associated virus injection. Mice in CFA group exhibited allodynia and anxiety-like behaviors. LPS induced neuroinflammation in N9 cells. Both CFA and LPS increased the levels of IBA1, ROS, and inflammatory markers. RUS (10 mg/kg in vivo and 1 μM in vitro) alleviated these alterations through NF-κB/MAPKs/NLRP3 signaling pathways but had no effect on pain hypersensitivity. TLR4 strongly interacted with RUS, and TLR4 overexpression abolished the effects of RUS on anxiety and neuroinflammation. RUS exerts anti-inflammatory and anxiolytic effects via TLR4-mediated NF-κB/MAPKs/NLRP3 signaling pathways, which provides a basis for the treatment of chronic pain-induced anxiety.
长期炎症可通过敏化中枢神经系统引起慢性疼痛和触发患者焦虑。然而，用于治疗慢性疼痛引起的焦虑的有效且副作用少的药物仍然缺乏。本研究评估了麦冬中重要活性化合物鲁斯可苷（RUS）对慢性炎症性疼痛和 N9 细胞小鼠模型的抗焦虑和抗炎作用。在 CFA 注射后，RUS（5、10 或 20mg/kg/天，ig）每天给药 1 次，共 7 天；评估小鼠的疼痛和焦虑样行为。评估 LPS 处理后 RUS（0.1、1、10μM）对 N9 小胶质细胞的抗炎作用。采用 qPCR 测定炎症标志物（TNF-α、IL-1β、IL-6、CD86、IL-4、ARG-1 和 CD206）。通过 Western blot 或免疫荧光染色检测 IBA1、ROS、NF-κB、TLR4、P-IKK、P-IκBα 和 P65、MAPKs（ERK、JNK 和 P38）、NLRP3（caspase-1、ASC 和 NLRP3）的水平。通过分子对接和腺相关病毒注射验证 RUS 的潜在靶标。CFA 组小鼠表现出痛觉过敏和焦虑样行为。LPS 在 N9 细胞中诱导神经炎症。CFA 和 LPS 均增加 IBA1、ROS 和炎症标志物的水平。RUS（体内 10mg/kg 和体外 1μM）通过 NF-κB/MAPKs/NLRP3 信号通路减轻这些改变，但对疼痛过敏无影响。TLR4 与 RUS 强烈相互作用，TLR4 过表达消除了 RUS 对焦虑和神经炎症的作用。RUS 通过 TLR4 介导的 NF-κB/MAPKs/NLRP3 信号通路发挥抗炎和抗焦虑作用，为治疗慢性疼痛引起的焦虑提供了依据。
