目的 利用网络药理学分析预测荣筋拈痛方调控巨噬细胞 M1 型极化治疗膝骨关节炎性滑膜炎的有效成 分、潜在作用靶点及核心信号通路，再通过体内外实验验证。方法 从 TCMSP 数据库中筛选荣筋拈痛方的功 能成分，检索 STITCH、Open Targets Platform 和 DrugBank 数据库，通过基因芯片分析筛选荣筋拈痛方调控滑 膜巨噬细胞极化治疗膝骨关节炎的潜在靶点和信号通路。运用免疫荧光、Western Blot 和 ELISA 等方法进行体 内外实验验证。结果 荣筋拈痛方调控巨噬细胞极化治疗膝骨关节炎的潜在靶点共 36 个。PPI 网络及基因富 集分析表明荣筋拈痛方可能是通过抑制 NF-κB 信号通路调控巨噬细胞的极化，从而对膝骨关节炎性滑膜炎起 治疗作用。动物实验中，HE 结果显示，与空白组比较，模型组小鼠滑膜较空白组显著增厚、滑膜下细胞的浸 润增加、血管翳的增生显著；与模型组比较，治疗组及对照组均有滑膜厚度变薄，滑膜内面浸润细胞减少及增 生血管翳减少。ELISA 结果显示，与空白组比较，模型组滑膜组织中 TNF-α 含量明显增加（P＜0.01）；与模型 组比较，治疗组 TNF-α 含量显著下降（P＜0.01）。Western Blot 结果显示，与空白组比较，模型组 p-NF-κB p65 显著激活（P＜0.01），治疗组可明显抑制 p-NF-κB p65 表达（P＜0.05）。体外实验中，免疫荧光结果显示， 与空白组比较，模型组的 p-NF-κB p65、CD16/32 蛋白表达明显增强；与模型组比较，治疗组和抑制剂组的 p-NF-κB p65、CD16/32 蛋白表达明显减弱。Western Blot 结果显示，与空白组比较，模型组 TNF-α 蛋白表达 增加（P＜0.01）；与模型组比较，治疗组及抑制剂组可降低 M1 型巨噬细胞中 TNF-α 蛋白表达（P＜0.05，P＜ 0.01）。与空白组比较，模型组可显著激活巨噬细胞 NF-κB p65 磷酸化（P＜0.01）；治疗组与抑制剂组均对 NF-κB p65 总蛋白表达虽没有明显影响，但与模型组比较，治疗组（P＜0.05）及抑制剂组（P＜0.01）可明显降低 巨噬细胞 NF-κB p65 的磷酸化。结论 荣筋拈痛方可能通过 NF-κB 信号通路调控滑膜巨噬细胞极化治疗膝骨 关节炎性滑膜炎。

Objective To predict the effective components，potential targets and core signaling pathways of Rongjin Niantong Fang （RJNTF） in regulating macrophage M1 polarization in the treatment of knee osteoarthritis synovitis by network pharmacology analysis，then verified by in vitro and in vivo experiments. Methods The functional components of RJNTF were screened from the TCMSP database， and then the databases STITCH， Open Targets Platform and DrugBank were searched. The potential targets and signaling pathways of Rongjin Niantong Fang in regulating synovial macrophage polarization in the treatment of knee osteoarthritis were screened by gene chip analysis. Immunofluorescence， Western Blot and ELISA were used for verification. Results There are 36 potential targets of RJNTF in regulating macrophage polarization for the treatment of knee osteoarthritis. PPI network and gene enrichment analysis showed that RJNTF may regulate the polarization of macrophages by inhibiting NF-κB signaling pathway，thus playing a therapeutic role in knee osteoarthritis synovitis. In the animal experiment，HE results showed that compared with the blank group，the synovial membrane of the model group was significantly thicker than that of the blank group， the infiltration of synovial cells was increased，and the proliferation of pannus was significant. Compared with the model group，both the treatment group and the control group had thinner synovial thickness，less infiltrating cells on the inner surface of the synovial membrane and less proliferative pannus. ELISA results showed that compared with the blank group， the content of TNF- α in the synovial tissue of the model group was significantly increased （P＜0.01）. Compared with the model group，the content of TNF-α in the treatment group was significantly decreased （P＜0.01）. Western Blot results showed that compared with the blank group，p-NF-κB p65 was significantly activated in the model group （P＜0.01），and the expression of p-NF-κB p65 was significantly inhibited in the treatment group （P＜0.05）. In vitro experiments，immunofluorescence results showed that compared with the blank group，the protein expressions of p-NF-κB p65 and CD16 / 32 in the model group were significantly enhanced. Compared with the model group，the protein expressions of p-NF- κB p65 and CD16/32 in the treatment group and the inhibitor group was significantly decreased. Western Blot results showed that compared with the blank group，the protein expression of TNF-α in the model group increased （P＜0.01）. Compared with the model group，the treatment group and the inhibitor group could reduce the protein expression of TNF-α in M1 macrophages （P＜0.05，P＜0.01）. Compared with the blank group， the model group could significantly activate the phosphorylation of NF-κB p65 in macrophages （P＜0.01）. There was no significant effect on the total protein expression of NF- κB p65 in the treatment group and the inhibitor group，but compared with the model group， the treatment group （P＜0.05） and the inhibitor group （P＜0.01） could significantly reduce the phosphorylation of macrophage NF-κB p65. Conclusion RJNTF may regulate synovial macrophage polarization through NF-κB signaling pathway to treat knee osteoarthritis synovitis.

R285.5

国家自然科学基金项目（82474553）；福建省自然科学基金面上项目（2022J01843）；国家中医药管理局高水平中医药重点学科建设项目 （中医骨伤科学）（zyyzdxk-2023106）；福建中医药大学中医骨伤科学学科开放课题资助（XGS2023004）；福建省中青年教师教育科研项目 （JAT210138）。