目的 探究雷公藤甲素對類風(fēng)濕關(guān)節(jié)炎成纖維樣滑膜細胞（FLSs）線粒體自噬、NOD樣受體蛋白3（NLRP3）炎癥小體活化和細胞焦亡的影響。方法 將FLSs細胞和類風(fēng)濕關(guān)節(jié)炎FLSs細胞進行傳代培養(yǎng)。將類風(fēng)濕性關(guān)節(jié)炎FLSs細胞采用0、10、20、40 ng/mL雷公藤甲素進行處理。采用蛋白質(zhì)印跡法（Western blotting）檢測線粒體自噬相關(guān)蛋白LC3B、p62、PHB2、NLRP3的蛋白表達。采用酶聯(lián)免疫吸附試驗（ELISA）檢測炎癥因子白細胞介素（IL）-1β和IL-18的水平。采用流式細胞術(shù)檢測細胞焦亡情況，并采用Western blotting法檢測焦亡相關(guān)蛋白GSDMD和GSDMD-N表達。結(jié)果 采用10、20、40 ng/mL雷公藤甲素處理類風(fēng)濕關(guān)節(jié)炎FLSs后，雷公藤甲素各劑量組均能顯著減弱類風(fēng)濕關(guān)節(jié)炎FLSs細胞的線粒體自噬相關(guān)蛋白LC3B的表達，上調(diào)p62和PHB2的表達；NLRP3表達水平均顯著降低，IL-1β和IL-18水平顯著降低；細胞焦亡率顯著降低；GSDMD蛋白表達顯著上升，GSDMD-N蛋白表達下降（P＜0.05、0.01、0.001），且呈劑量相關(guān)性。結(jié)論 雷公藤甲素能通過抑制線粒體自噬、NLRP3炎癥小體激活和細胞焦亡，抑制細胞炎癥反應(yīng)。

Objective To investigate the effects of triptolide on mitochondrial autophagy, NLRP3 inflammatome activation and pyroptosis of fibroblast synovial cells (FLSs) in rheumatoid arthritis. Methods FLSs cells and rheumatoid arthritis FLSs cells were cultured by passage. Rheumatoid arthritis FLSs cells were treated with 0, 10, 20, 40 ng/mL triptolide. The protein expressions of mitochondrial autophagy related proteins LC3B, p62, PHB2, and NLRP3 were detected by Western blotting. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory cytokines IL-1β and IL-18. Pyroptosis was detected by flow cytometry, and the expressions of pyroptosis related proteins GSDMD and GSDMD-N were detected by Western blotting. Results After treatment with 10, 20, 40 ng/mL triptolide, the expression of mitochondrial autophagy associated protein LC3B in FLSs cells of rheumatoid arthritis was significantly reduced in each dose group, and the expression of p62 and PHB2 was upregulated. Expression levels of NLRP3, IL-1β, and IL-18 were significantly decreased. Pyrodeath rate was significantly reduced. Expression of GSDMD protein was significantly increased, and the expression of GSDMD-N protein was decreased in a dose-dependent manner (P < 0.05, 0.01, 0.001). Conclusion Triptolide can inhibit cellular inflammation by inhibiting mitochondrial autophagy, activation of NLRP3 inflammatome and pyroptosis.

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