| 孙利平,宋亚玲,李春艳.阿米卡星调控微小 RNA-223对脂多糖诱导的肺泡上皮细胞凋亡以及肿瘤坏死因子 α、白细胞介素 6表达的影响[J].安徽医药,2022,26(7):1310-1314. |
| 阿米卡星调控微小 RNA-223对脂多糖诱导的肺泡上皮细胞凋亡以及肿瘤坏死因子 α、白细胞介素 6表达的影响 |
| Effects of amikacin on apoptosis, TNF-α and IL-6 expression of effects of amikacin on LPS-induced apoptosis of alveolar epithelial cells and the expression of tumor necrosis factor-α and interleukin-6 by regulating miR-223 |
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| DOI：10.3969/j.issn.1009-6469.2022.07.009 |
| 中文关键词: 阿米卡星 微小 RNA-223 肺泡上皮细胞 细胞凋亡 炎症反应 急性肺损伤 |
| 英文关键词: Amikacin miR-223 Alveolar epithelial cells Apoptosis Inflammatory response Acute lung injury |
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| 中文摘要: |
| 目的探讨阿米卡星对脂多糖( LPS)诱导的肺泡上皮细胞凋亡以及肿瘤坏死因子 α(TNF-α)、白细胞介素( IL)-6表达的影响和分子机制。方法该研究于 2020年 1—7月完成，采用 LPS诱导肺泡上皮细胞，构建急性肺损伤细胞模型。将肺泡上皮细胞分为对照组、模型组( LPS处理 24 h)、实验组(由 LPS和不同剂量的阿米卡星处理 24 h)、 anti-miR-NC组(转染微小 RNA阴性对照后进行 LPS处理)、 anti-miR-223组(转染微小 RNA-223抑制剂后进行 LPS处理)、实验 3+miR-NC组(转染微小 RNA阴性对照后进行 LPS和 15 μg/L阿米卡星处理)、实验 3+miR-223组(转染微小 RNA-223激动剂后进行 LPS和 15 μg/L阿米卡星处理)。流式细胞术检测细胞凋亡；蛋白质印迹法( Western blotting)检测兔源裂解的胱天蛋白酶 3(cl-caspase3)表达。酶联免疫吸附测定( ELISA)检测细胞上清液中 TNF-α和 IL-6含量。实时荧光定量 PCR(RT-qPCR)检测 miR-223(微小 RNA-223)表达。结果与对照组比较，模型组肺泡上皮细胞凋亡率( 25.81±1.82)%、cl-caspase3(0.81±0.06)和 miR-223表达( 0.17±0.02)、细胞上清液中 TNF-α(793.92±34.49)ng/L和 IL-6含量( 412.04±23.94)ng/L升高( P <0.05)。与模型组比较，实验组肺泡上皮细胞凋亡率 |
| 英文摘要: |
| Objective To investigate the effect and molecular mechanism of amikacin on the apoptosis, tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) expression of lipopolysaccharide (LPS)-induced alveolar epithelial cells.Methods The experimentwas completed from January to July 2020, LPS was used to induce alveolar epithelial cells to construct an acute lung injury cell model.Alveolar epithelial cells were assigned into control group, model group (treated with LPS for 24-hour), experimental group (treated with LPS and different doses of amikacin for 24-hour), and anti-miR-NC group (transfected with microRNA negative control followed by LPS treatment), anti-miR-223 group (transfected with microRNA-223 inhibitor and then performed LPS treatment), experiment 3+miR-NC group (transfected with microRNA-223 inhibitor) LPS and 15 μg/L amikacin treatment after microRNA negative control), experiment 3+ miR-223 group (LPS and 15 μg/L amikacin treatment after microRNA-223 agonist transfection). Flow cytometry detected apoptosis; Western blotting detected cl-caspase 3 expression. Enzyme-linked immunosorbent assay (ELISA) kit detected the contents of TNF-α and IL-6 in cell supernatants. Real-time quantitative PCR (RT-qPCR) detected miR-223 expression.Results Compared with the con-trol group, the apoptosis rate of alveolar epithelial cells in the model group was (25.81±1.82)%, the expression of cl-caspase3 (0.81± 0.06) and miR-223 (0.17±0.02), TNF-α in cell supernatant (793.92±34.49) ng/L and IL-6 content (412.04±23.94) ng/L increased, and the difference between groups was statistically significant (P<0.05). Compared with the model group, the apoptosis rate of alveolar epi-thelial cells in the experimental group (22.07±1.69)%, the expression of cl-caspase 3 (0.68±0.05) and miR-223 (0.28±0.03), the con-tents of TNF-α (478.38±25.85) ng/L and IL-6 (202.95±15.55) ng/L in the cell supernatant were decreased, and the difference between groups was statistically significant (P<0.05). Compared with the anti-miR-NC group, the apoptosis rate of alveolar epithelial cells in the anti-miR-223 group was (10.12±1.13)%, the expression of cl-caspase 3 (0.27±0.03) and miR-223 (1.69±0.08), The contents of TNF-α (403.48±29.94) ng/L and IL-6 (166.35±15.61) ng/L in the cell supernatant decreased, and the difference between groups was statistical-ly significant (P<0.05). Compared with experiment 3+miR-NC group, the apoptosis rate of alveolar epithelial cells in experiment 3+ miR-223 group was (24.03±1.64)%, the expression of cl-caspase 3 (0.75±0.06) and miR-223 (4.67±0.22), The contents of TNF-α (726.18±33.57) ng/L and IL-6 (385.07±22.15) ng/L in the cells supernatant were increased, and the difference between the groups was statistically significant (P<0.05).Conclusions Amikacin has obvious anti-inflammatory and anti-apoptotic effects on LPS-induced al-veolar epithelial cells, and its mechanism may be related to the inhibition of miR-223 expression. |
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