目的 基于核因子-κB（nuclear factor-κB，NF-κB）信号通路研究千斤拔总黄酮（total flavonoids of Moghania philippinensis，MPTF）抗人类风湿性关节炎成纤维样滑膜细胞（rheumatoid arthritis-fibroblast-like synoviocyte，RA-FLS）炎症的体外作用机制。方法 采用肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）诱导RA-FLS炎症模型，检测细胞活性及白细胞介素-1β（interleukin-1β，IL-1β）、IL-6、IL-8、基质金属蛋白酶3（matrix metalloproteinase 3，MMP3），MMP9，p62，Beclin-1和微管相关蛋白1轻链3（microtubule-associated protein 1 light chain 3，LC3）的蛋白表达，探究MPTF的抗炎作用和对自噬的影响。通过使用自噬流抑制剂和激动剂来检测不同自噬流量对抗炎作用的潜在作用。通过Western blotting检测NF-κB抑制因子α（inhibitor of NF-κB-α，IκBα）的蛋白表达及p-p65/p65的值，并利用其抑制剂和激动剂探究MPTF是否通过NF-κB信号通路介导的自噬发挥抗炎作用。结果 在TNF-α诱导的RA-FLS炎症模型中，MPTF可以提高细胞活力（P＜0.05、0.01），显著降低IL-1β、IL-6、IL-8、MMP3、MMP9和p62的蛋白表达水平（P＜0.05、0.01），提高Beclin-1的蛋白表达水平和LC3II/LC3Ⅰ的值（P＜0.05、0.01）；且MPTF可以显著提高IκBɑ的蛋白表达水平（P＜0.01），降低p-p65/p65的值（P＜0.05、0.01）。结论 MPTF通过抑制NF-κB信号通路增强自噬，从而发挥抗RA-FLS炎症的作用。

Objective To investigate the mechanism of the total flavonoids of Moghania philippinensis (MPTF) in vitro from the perspectives of human rheumatoid arthritis-fibroblast-like synoviocyte (RA-FLS) inflammation based on the nuclear factor-κB (NF-κB) signaling pathway. Methods The RA-FLS inflammation model was induced by tumor necrosis factor-α (TNF-α). The cell viability and the expressions of interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), matrix metalloproteinase 3 (MMP3), MMP9, p62, Beclin-1 and microtubule-associated protein 1 light chain 3 (LC3) proteins were determined to assess the anti-inflammatory capabilities and the effects on autophagy of MPTF. The potential impact of different autophagy fluxes on the anti-inflammatory effects was detected by using the inhibitor and the agonist. Finally, the expressions of inhibitor of NF-κB-α (IκBα) and p-p65/p65 proteins were examined by Western blotting, and the inhibitor and agonist were utilized to make an inquiry of whether MPTF exerts anti-inflammatory effects through the autophagy mediated by the NF-κB signaling pathway. Results MPTF could improve cell viability (P < 0.05, 0.01), significantly reduce the expression of IL-1β、IL-6、IL-8、MMP3、MMP9 and p62 (P < 0.05, 0.01), improve the expression of Beclin-1 and LC3II/LC3Ⅰ (P < 0.05, 0.01). Moreover, MPTF could improve the expression of IκBɑ (P < 0.01), and reduce the expression of p-p65/p65 (P < 0.05, 0.01). Conclusion MPTF can increase autophagy by inhibiting the NF-κB signaling pathway, thereby exerting an anti-RA-FLS inflammatory effect.

R285.5

广东省基础与应用基础研究基金-省企联合基金面上项目（2023A1515220188）