目的 探討毛蕊花糖苷對棕櫚酸處理的HepG2細胞線粒體保護作用，并探討其作用機制。方法 將HepG2細胞設置為對照組、棕櫚酸（100 μmol/L）組、毛蕊花糖苷（50 μmol/L）組、棕櫚酸（100 μmol/L）＋毛蕊花糖苷（50 μmol/L）組。MitoSox熒光探針在激光共聚焦顯微鏡下檢測線粒體超氧化物水平的變化；JC-1熒光探針在激光共聚焦顯微鏡下觀察線粒體膜電位的改變；Tomm20、微管相關蛋白輕鏈3（LC3）特異性抗體進行免疫熒光染色，觀察共定位情況；線粒體提取試劑盒提取HepG2細胞內線粒體，蛋白質免疫印跡技術檢測線粒體上LC-3Ⅱ蛋白相對表達水平的變化。結果 棕櫚酸組細胞內MitoSox紅色熒光顯著增強（P＜0.05），棕櫚酸＋毛蕊花糖苷組HepG2細胞內紅色熒光強度顯著降低（P＜0.05）；棕櫚酸組HepG2細胞內綠/紅熒光強度比例顯著升高（P＜0.05），棕櫚酸＋毛蕊花糖苷組HepG2細胞內綠/紅熒光強度比例顯著降低（P＜0.05）；Tomm20和LC3共定位分析結果顯示，棕櫚酸組線粒體與LC3在空間上沒有明顯的共定位，棕櫚酸＋毛蕊花糖苷組HepG2細胞內線粒體形態(tài)完整，線粒體與LC3顯示出明顯的共定位；蛋白質免疫印跡結果顯示，與棕櫚酸組比較，棕櫚酸＋毛蕊花糖苷組HepG2細胞內線粒體上LC3-Ⅱ蛋白相對表達水平顯著升高（P＜0.05）。結論 在棕櫚酸處理的HepG2細胞模型中，毛蕊花糖苷可能通過誘導線粒體自噬發(fā)揮線粒體保護作用。

Objective To investigate the protective effect of acteoside on mitochondria of HepG2 cells treated with palmitic acid, and to explore its mechanism. Methods HepG2 cells were set as the control group, palmitic acid (100 μmol/L) group, acteoside (50 μmol/L) group, and palmitic acid (100 μmol/L) + acteoside (50 μmol/L) group. Changes in mitochondrial superoxide levels were detected by MitoSox fluorescent probes under a laser confocal microscope. The JC-1 fluorescent probe was used to observe the changes of mitochondrial membrane potential under a laser confocal microscope. Immunofluorescence staining was performed with Tomm20 and LC3 specific antibodies to observe the co-localization situation. Mitochondria were extracted from HepG2 cells using the mitochondrial extraction kit, and the changes in the relative expression level of LC-3Ⅱ protein on mitochondria were detected by Western blotting. Results The red fluorescence of MitoSox in the cells of the palmitic acid group was significantly enhanced (P < 0.05), while the intensity of red fluorescence in the cells of HepG2 in the palmitic acid + acteoside group was significantly decreased (P < 0.05). The ratio of intracellular green/red fluorescence intensity in HepG2 cells in the palmitic acid group was significantly increased (P < 0.05), while the ratio of intracellular green/red fluorescence intensity in HepG2 cells in the palmitic acid + acteoside group was significantly decreased (P < 0.05). The results of Tomm20 and LC3 co-localization analysis showed that there was no obvious co-localization of mitochondria and LC3 in space in the palmitic acid group, while the mitochondria morphology in HepG2 cells was intact, and mitochondria and LC3 showed obvious co-localization. The results of Western blotting showed that the relative expression levels of LC3-II protein on mitochondria in HepG2 cells in the palmitic acid + acteoside group were significantly increased compared with those in the palmitic acid group (P < 0.05). Conclusion In HepG2 cells treated with palmitic acid, acteoside may exert mitochondrial protection by inducing mitochondrial autophagy.

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新疆維吾爾自治區(qū)自然科學基金資助項目（2022D01B41）；新疆維吾爾醫(yī)學?？茖W校自然科學基金重點實驗室開放課題項目（SYS2024-002）