目的 基于核因子-κB（nuclear factor-κB，NF-κB）信號(hào)通路研究千斤拔總黃酮（total flavonoids of Moghania philippinensis，MPTF）抗人類風(fēng)濕性關(guān)節(jié)炎成纖維樣滑膜細(xì)胞（rheumatoid arthritis-fibroblast-like synoviocyte，RA-FLS）炎癥的體外作用機(jī)制。方法 采用腫瘤壞死因子-α（tumor necrosis factor-α，TNF-α）誘導(dǎo)RA-FLS炎癥模型，檢測(cè)細(xì)胞活性及白細(xì)胞介素-1β（interleukin-1β，IL-1β）、IL-6、IL-8、基質(zhì)金屬蛋白酶3（matrix metalloproteinase 3，MMP3），MMP9，p62，Beclin-1和微管相關(guān)蛋白1輕鏈3（microtubule-associated protein 1 light chain 3，LC3）的蛋白表達(dá)，探究MPTF的抗炎作用和對(duì)自噬的影響。通過(guò)使用自噬流抑制劑和激動(dòng)劑來(lái)檢測(cè)不同自噬流量對(duì)抗炎作用的潛在作用。通過(guò)Western blotting檢測(cè)NF-κB抑制因子α（inhibitor of NF-κB-α，IκBα）的蛋白表達(dá)及p-p65/p65的值，并利用其抑制劑和激動(dòng)劑探究MPTF是否通過(guò)NF-κB信號(hào)通路介導(dǎo)的自噬發(fā)揮抗炎作用。結(jié)果 在TNF-α誘導(dǎo)的RA-FLS炎癥模型中，MPTF可以提高細(xì)胞活力（P＜0.05、0.01），顯著降低IL-1β、IL-6、IL-8、MMP3、MMP9和p62的蛋白表達(dá)水平（P＜0.05、0.01），提高Beclin-1的蛋白表達(dá)水平和LC3II/LC3Ⅰ的值（P＜0.05、0.01）；且MPTF可以顯著提高IκBɑ的蛋白表達(dá)水平（P＜0.01），降低p-p65/p65的值（P＜0.05、0.01）。結(jié)論 MPTF通過(guò)抑制NF-κB信號(hào)通路增強(qiáng)自噬，從而發(fā)揮抗RA-FLS炎癥的作用。

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

廣東省基礎(chǔ)與應(yīng)用基礎(chǔ)研究基金-省企聯(lián)合基金面上項(xiàng)目（2023A1515220188）