摘要目的:探讨黄芪甲苷对脂多糖诱导的人鼻上皮细胞炎症反应与线粒体功能障碍的缓解作用，并初步揭示其潜在机制。方法:根据干预措施将人鼻腔上皮RPMI 2650细胞分为4组：对照组（磷酸盐缓冲液处理24 h）、脂多糖组（10 μg/ml脂多糖处理24 h）、脂多糖＋10 μmol/L黄芪甲苷组（10 μg/ml脂多糖联合10 μmol/L黄芪甲苷共同处理24 h）、脂多糖＋20 μmol/L黄芪甲苷组（10 μg/ml脂多糖联合20 μmol/L黄芪甲苷共同处理24 h）。通过细胞计数试剂盒-8实验检测细胞活力；通过酶联免疫吸附试验检测肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-6和IL-10水平；采用2′,7′-二氯二氢荧光素二乙酸酯荧光探针法检测活性氧水平；通过JC-1线粒体膜电位检测试剂盒检测线粒体膜电位；通过蛋白质印迹法检测基质金属蛋白酶3（MMP3）、含有血小板反应蛋白基序的解聚素样金属蛋白酶（ADAMTS5）、Ⅱ型胶原蛋白（Collagen Ⅱ）、核转录因子红系2相关因子2（Nrf2）、Kelch样ECH关联蛋白1（Keap1）和血红素加氧酶-1（HO-1）蛋白的相对表达量。组间比较采用单因素方差分析和Tukey HSD事后检验。结果:脂多糖＋10 μmol/L黄芪甲苷组的细胞活力，IL-10水平，Collagen Ⅱ、Nrf2、HO-1蛋白的相对表达量[（68.00±4.36）%、（116.00±12.53）pg/ml、0.36±0.06、0.51±0.07、0.20±0.05]均高于脂多糖组[（48.67±4.51）%、（48.00±6.56）pg/ml、0.15±0.04、0.32±0.04、0.04±0.03]，TNF-α、IL-6水平，MMP3、ADAMTS5、Keap1蛋白的相对表达量[（561.00±30.12、504.30±31.53）pg/ml、0.77±0.05、0.69±0.04、0.38±0.06]则均低于脂多糖组[（771.00±35.68、626.00±27.51）pg/ml、1.00±0.10、1.00±0.09、1.00±0.10]，差异有统计学意义（ P<0.05、0.01）；而活性氧水平[（88.00±8.00）%]虽也低于脂多糖组[（97.33±7.51）%]，但差异没有统计学意义（ P>0.05）。脂多糖＋20 μmol/L黄芪甲苷组的细胞活力，IL-10水平，Collagen Ⅱ、Nrf2、HO-1蛋白的相对表达量[（83.33±5.51）%、（149.70±13.20）pg/ml、0.69±0.04、0.92±0.06、0.37±0.06]均高于脂多糖组，TNF-α、IL-6、活性氧水平，MMP3、ADAMTS5、Keap1蛋白的相对表达量[（324.30±21.55、384.30±18.01）pg/ml、（10.33±3.06）%、0.36±0.06、0.31±0.06、0.38±0.14]则均低于脂多糖组，差异均有统计学意义（均 P<0.01）。 结论:黄芪甲苷可通过激活Nrf2/HO-1信号通路，缓解脂多糖诱导的人鼻上皮细胞炎症反应与线粒体功能障碍。

abstractsObjective:To investigate the alleviating effects of astragaloside Ⅳ on lipopolysaccharide-induced inflammatory response and mitochondrial dysfunction in human nasal epithelial cells, and to preliminarily elucidate its underlying mechanisms.Methods:Human nasal epithelial RPMI 2650 cells were divided into 4 groups based on intervention: the control group (treated with phosphate buffered saline for 24 h), the lipopolysaccharide group (treated with 10 μg/ml lipopolysaccharide for 24 h), the lipopolysaccharide+10 μmol/L astragaloside Ⅳ group (treated with 10 μg/ml lipopolysaccharide combined with 10 μmol/L astragaloside Ⅳ for 24 h), and the lipopolysaccharide+20 μmol/L astragaloside Ⅳ group (treated with 10 μg/ml lipopolysaccharide combined with 20 μmol/L astragaloside Ⅳ for 24 h). The cell viability was detected by cell counting kit-8 assay. The levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and IL-10 were detected by enzyme-linked immunosorbent assay. The level of reactive oxygen species was detected by 2′,7′-dichlorodihydrofluorescein diacetate fluorescent probe method. Mitochondrial membrane potential was detected by JC-1 mitochondrial membrane potential detection kit. The relative expression levels of matrix metalloproteinase 3 (MMP3), a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS5), type Ⅱ Collagen (Collagen Ⅱ), nuclear factor-erythroid 2-related factor 2 (Nrf2), Kelch-like ECH-associated protein 1 (Keap1) and heme oxygenase-1 (HO-1) proteins were detected by Western blotting assay. One-way analysis of variance and Tukey HSD post hoc test were used for comparison between groups.Results:The cell viability, and the level of IL-10, and the relative expression levels of Collagen Ⅱ, Nrf2, and HO-1 proteins in the lipopolysaccharide+10 μmol/L astragaloside Ⅳ group [(68.00±4.36)%, (116.00±12.53) pg/ml, 0.36±0.06, 0.51±0.07, and 0.20±0.05] were higher than those in the lipopolysaccharide group [(48.67±4.51)%, (48.00±6.56) pg/ml, 0.15±0.04, 0.32±0.04, and 0.04±0.03], while the levels of TNF-α and IL-6, and the relative expression levels of MMP3, ADAMTS5, and Keap1 proteins [(561.00±30.12, 504.30±31.53) pg/ml, 0.77±0.05, 0.69±0.04, and 0.38±0.06] were lower than those in the lipopolysaccharide group [(771.00±35.68, 626.00±27.51) pg/ml, 1.00±0.10, 1.00±0.09, and 1.00±0.10], and the differences were statistically significant ( P<0.05, 0.01). The level of reactive oxygen species [(88.00±8.00)%] was also lower than that in the lipopolysaccharide group [(97.33±7.51)%], but the difference was not statistically significant ( P>0.05). The cell viability, the level of IL-10, and relative expression levels of Collagen Ⅱ, Nrf2 and HO-1 proteins in the lipopolysaccharide+20 μmol/L astragaloside Ⅳ group [(83.33±5.51)%, (149.70±13.20) pg/ml, 0.69±0.04, 0.92±0.06, and 0.37±0.06] were higher than those in the lipopolysaccharide group, while the levels of TNF-α, IL-6, and reactive oxygen species, and the relative expression levels of MMP3, ADAMTS5 and Keap1 proteins [(324.30±21.55, 384.30±18.01) pg/ml, (10.33±3.06)%, 0.36±0.06, 0.31±0.06, and 0.38±0.14] were lower than those in the lipopolysaccharide group, and the differences were statistically significant (all P<0.01). Conclusions:Astragaloside Ⅳ may alleviate lipopolysaccharide-induced inflammatory response and mitochondrial dysfunction in human nasal epithelial cells by activating the Nrf2/HO-1 signaling pathway.