目的 探討橄欖苦苷對脂多糖（LPS）誘導的小鼠腹腔巨噬細胞（RAW264.7）炎癥的保護作用及機制。方法 MTT法檢測橄欖苦苷（0、10、20、40 μmol/L）對RAW264.7細胞活性的影響；用橄欖苦苷（10、20、40 μmol/L）預處理細胞1 h后，LPS誘導炎癥模型，Griess試劑檢測細胞內Nitrite釋放；Western blotting方法檢測細胞iNOS、COX-2、Nrf2、Keap-1、HO-1、NQO1蛋白表達水平；流式細胞儀檢測細胞內NO、ROS、Ca²⁺的水平；熒光顯微鏡檢測線粒體膜電位（MMP）水平；ELISA法檢測細胞上清液中腫瘤壞死因子-α（TNF-α）、白介素-6（IL-6）的釋放。結果 與模型組比較，橄欖苦苷組Nitrite釋放水平顯著降低（P<0.05、0.01、0.001），iNOS、COX-2蛋白的表達顯著降低（P<0.001），NO的產(chǎn)生顯著減少（P<0.05、0.01）；TNF-α、IL-6的釋放受到顯著抑制（P<0.001）；Ca²⁺釋放受到顯著抑制（P<0.01）；ROS生成受到顯著抑制（P<0.01）；JC-1單體降低，恢復聚合物狀態(tài)（紅色熒光變多），MMP穩(wěn)定性增強；Keap1蛋白表達顯著降低， Nrf2、HO-1、NQO1蛋白的表達均顯著升高（P<0.05、0.01、0.001）。結論 橄欖苦苷對LPS誘導的RAW264.7細胞炎癥具有抑制作用，其作用機制可能與激活Nrf2/HO-1信號通路相關。

Objective To explore the anti-inflammatory activity of Oleuropein in LPS-stimulated RAW264.7 cells via Nrf2/HO-1 signaling pathway. Methods MTT method was used to detect the effect of oleuropein (0, 10, 20, 40 μmol/L) on the activity of RAW264.7 cells. After being pretreated with oleuropein (10, 20, 40 μmol/L) for one hour, LPS induced inflammation model was established. Griess reagent was used to detect the release of nitrite in cells. Western blotting method was used to detect the expression of iNOS, COX-2, Nrf2, keap-1, HO-1 and NQO1 in cells. Flow cytometry was used to detect the expression of NO, ROS, and Ca²⁺ in cells. The levels of MMP, TNF-α and IL-6 in supernatant were detected by ELISA. Results Compared with model group, the release level of nitrite in the oleuroside group decreased significantly (P<0.05, 0.01, 0.001), the expression of iNOS and COX-2 decreased significantly (P<0.001), the production of NO decreased significantly (P<0.05, 0.01), the release of TNF-α and IL-6 was inhibited significantly (P<0.001), and the release of Ca2 + was inhibited significantly (P<0.01); ROS production was significantly inhibited (P<0.01); JC-1 monomer decreased, polymer state was restored (more red fluorescence), MMP stability was enhanced; Keap1 protein expression was significantly decreased, Nrf2, HO-1, NQO1 protein expression were significantly increased (P<0.05, 0.01 and 0.001). Conclusion Oleuropein has a significant inhibitory effect on LPS-induced RAW264.7 cells inflammatory response, which reduces the inflammatory process by activating the Nrf2/HO-1 signaling pathway.

R962.2

國家青年科學自然基金項目（81803807）；廣西自治區(qū)青年科學基金項目（2018JJB140265）；廣西高校引進海外高層次人才“百人計劃”項目（05018064）；廣西科技基地和人才專項（2018AD19034）。