目的 基于核苷酸结合寡聚化结构域样受体蛋白3（NOD-like receptor family pyrin domain containing 3，NLRP3）/白细胞介素-1β（interleukin-1β，IL-1β）通路探讨筋骨止痛凝胶改善膝骨性关节炎大鼠膝关节炎症、疼痛的作用和机制。方法 70只雄性SD大鼠随机分为对照组，模型组，筋骨止痛凝胶高、中、低剂量（1.24、0.62、0.31 g/kg）组，扶他林（0.105 g/kg）组和青鹏膏（0.105 g/kg）组，每组10只。对照组大鼠左膝关节腔注射50 μL无菌生理盐水，其余各组注射40 mg/mL单碘乙酸钠建立膝骨性关节炎大鼠模型，造模后1 d开始给药，持续14 d。测定各组大鼠机械痛阈值（paw withdrawal mechanical threshold，PWT）、热刺激缩足反射潜伏期（paw withdrawal thermal latency，PWTL）及足负重；采用苏木素-伊红（HE）染色观察膝关节滑膜组织病理情况；采用ELISA检测血清中IL-1β、前列腺素E₂（prostaglandin E₂，PGE₂）及关节液中IL-1β水平；采用qRT-PCR检测滑膜组织中IL-1β、PGE₂和NLRP3 mRNA表达。结果 与模型组比较，各给药组大鼠PWT、PWTL和足负重均显著升高（P＜0.05、0.01）；筋骨止痛凝胶低剂量组、扶他林组和青鹏膏组滑膜组织病理学评分显著降低（P＜0.01）；筋骨止痛凝胶高剂量组血清和关节液中IL-1β水平均明显降低（P＜0.05、0.01），筋骨止痛凝胶低、中剂量组和青鹏膏组血清中PGE₂水平明显降低（P＜0.05、0.01）；筋骨止痛凝胶低剂量组和扶他林组滑膜组织中IL-1β mRNA表达水平均显著降低（P＜0.05、0.01），筋骨止痛凝胶低、中剂量组及扶他林组和青鹏膏组PGE₂和NLRP3 mRNA表达水平均显著降低（P＜0.05、0.01）。结论 筋骨止痛凝胶可能通过调控IL-1β、PGE₂、NLRP3炎症因子的表达，抑制滑膜炎进展，从而发挥抗膝骨性关节炎的作用。

Objective To explore the effect and mechanism of Jingu Zhitong Gel (筋骨止痛凝胶) on knee joint inflammation and pain in rats with knee osteoarthritis (KOA) based on NOD-like receptor family pyrin domain containing 3 (NLRP3)/interleukin-1β (IL-1β) pathway. Methods Seventy male SD rats were randomly divided into control group, model group, Jingu Zhitong Gel high-, medium- and low-dose (1.24, 0.62, 0.31 g/kg) groups, voltarin (0.105 g/kg) group and Qingpeng Ointment (青鹏膏, 0.105 g/kg) group, with 10 rats in each group. Rats in control group were injected with 50 μL sterile physiological saline into left knee joint cavity, while rats in other groups were injected with 40 mg/mL sodium monoiodide to establish a rat model of KOA. Drugs were given 1 d after the model was established and lasted for 14 d. The mechanical pain threshold (PWT), paw withdrawal thermal latency (PWTL) and foot load of rats in each group were measured. HE staining was used to observe the pathological changes of synovial tissue of knee joint. The levels of IL-1β and prostaglandin E₂ (PGE₂) in serum and IL-1β in joint fluid were detected by ELISA. The expressions of IL-1β, PGE₂ and NLRP3 mRNA in synovial tissue were detected by qRT-PCR. Results Compared with model group, PWT, PWTL and foot load of rats in each group were significantly increased (P < 0.05, 0.01). The histopathological scores of synovium in Jingu Zhitong Gel low-dose group, votaline group and Qingpeng Ointment group were significantly decreased (P < 0.01). The levels of IL-1β in serum and joint fluid in Jingu Zhitong Gel high-dose group were significantly decreased (P < 0.05, 0.01), and level of PGE2 in serum in Jingu Zhitong Gel low-, medium-dose groups and Qingpeng Ointment group were significantly decreased (P < 0.05, 0.01). The expression level of IL-1β mRNA in synovial tissue of Jingu Zhitong Gel low-dose group and votaline group were significantly decreased (P < 0.05, 0.01), while the expression levels of PGE₂ and NLRP3 mRNA in Jingu Zhitong Gel low-, medium-dose groups, votaline group and Qingpeng Ointment group were significantly decreased (P < 0.05, 0.01). Conclusion Jingu Zhitong Gel may inhibit the progress of synovitis by regulating the expressions of IL-1β, PGE₂ and NLRP3 inflammatory factors, thus playing an anti-KOA role.

R285.5

2021年国家中医药管理局岐黄学者项目