| 温玲,李宝琪,赵艳敏,等.基于 NF-κB信号通路探讨阿奇霉素对脂多糖诱导肺泡上皮细胞增殖、凋亡的作用[J].安徽医药,2025,29(7):1303-1307. |
| 基于 NF-κB信号通路探讨阿奇霉素对脂多糖诱导肺泡上皮细胞增殖、凋亡的作用 |
| Effect of azithromycin on the proliferation and apoptosis of alveolar epithelial cells induced by lipopolysaccharide based on NF-κB signaling pathway |
| |
| DOI：10.3969/j.issn.1009-6469.2025.07.008 |
| 中文关键词: 阿奇霉素 肺泡上皮细胞 急性肺损伤 核因子 κB信号通路 增殖 凋亡 |
| 英文关键词: Azithromycin Alveolar epithelial cells Acute lung injury Nuclear transcription factor-κB signaling pathway Pro-liferation Apoptosis |
| 基金项目:秦皇岛市科学技术研究与发展计划项目( 202301A102) |
| 作者 | 单位 | E-mail | | 温玲 | 秦皇岛市第一医院,儿科 ICU,河北秦皇岛 066000 | | | 李宝琪 | 秦皇岛市第一医院,儿科 ICU,河北秦皇岛 066000 | pete1007@163.com | | 赵艳敏 | 秦皇岛市第一医院,药学部,河北秦皇岛 066000 | | | 郑舒扬 | 秦皇岛市第一医院,儿科 ICU,河北秦皇岛 066000 | | | 苏颖 | 秦皇岛市第一医院,儿科 ICU,河北秦皇岛 066000 | |
|
| 摘要点击次数: 863 |
| 全文下载次数: 226 |
| 中文摘要: |
| 目的探究阿奇霉素对脂多糖( LPS)诱导的人肺泡上皮细胞增殖、凋亡及核转录因子 -κB(NF-κB)信号通路的调控作用。方法 2023年 2—7月，体外培养人肺泡上皮细胞 A549，分为空白组、 LPS组、实验组( LPS+1、LPS+2、LPS+4、LPS+8)、阿奇霉素组、抑制剂组、阿奇霉素 +抑制剂组和阿奇霉素 +激活剂组。白细胞介素( IL)-6、IL-8及 IL-1β通过酶联免疫吸附分析检测；细胞计数试剂盒 -8(CCK-8)进行细胞活力检测； 5-乙炔基 -2'脱氧尿嘧啶核苷( EdU)进行增殖率检测； Hoechst 33258染色法测定细胞凋亡率；核转录因子 -κB p65(NF-κB p65)、磷酸化( p-)NF-κB p65、细胞周期素 D1(Cyclin D1)及胱天蛋白酶 3(caspase-3)蛋白表达水平检测使用蛋白质印迹法。结果 LPS组较空白组细胞 IL-6、IL-8和 IL-1β水平升高( P<0.05)活力降低( P<0.05)。与 LPS组相比， LPS+4和 LPS+8组 IL-6、IL-8、IL-1β表达水平降低［ LPS组:(40.24±4.59)ng/L、(249.53±8.60)n，g/L、(85.22±2.64) ng/L；LPS+4组:(23.23±2.86)ng/L、(170.99±10.62)ng/L、(25.87±2.91)ng/L；LPS+8组:(20.38±2.57)ng/L、(127.57±5.49)ng/L、(27.56±2.89)ng/L］(P<0.05)细胞活力升高［ LPS+4组:(57.77±4.24)%；LPS+8组:(71.80±7.40)%］(P<0.05)。该研究在 LPS组基础上，选择与之差异有统计学意，义且较低浓度的 4 mg/L阿奇霉素作为阿奇霉素组，并相应地分别加入 BAY 11-7082和 Prostratin进行 NF-κB通路验证实验。相较于空白组， LPS组增殖率、 CyclinD1蛋白水平下降，凋亡率、 caspase-3、p-NF-κB p65蛋白水平升高( P<0.05)。阿奇霉素组与抑制剂组中上述指标的变化有显著扭转( P<0.05)。与阿奇霉素组相比，阿奇霉素 +抑制剂组细胞增殖率、 CyclinD1蛋白表达水平进一步升高，细胞凋亡率、 Caspase-3、p-NF-κB p65蛋白表达水平进一步降低，阿奇霉素 +激活剂组则显著扭转了上述指标的变化( P<0.05)。结论阿奇霉素能够通过抑制 NF-κB信号通路发挥对 A549细胞炎症损伤的保护作用，促进细胞增殖并抑制凋亡。 |
| 英文摘要: |
| Objective To explore the regulation of azithromycin on proliferation and apoptosis of human alveolar epithelial cells in-duced by lipopolysaccharide (LPS) and nuclear transcription factor-κB (NF-κB) signaling pathway.Methods From February to July2023, human alveolar epithelial cells A549 were cultured in vitro and divided into blank group, LPS group, experimental group (LPS+1,LPS+2, LPS+4, LPS+8), azithromycin group, inhibitor group, azithromycin + inhibitor group and azithromycin + activator group. After24 h of intervention, the expression levels of inflammatory factors interleukin (IL)-6, IL-8 and IL-1β were determined by enzyme-linked immunosorbent assay. Cell count Kit 8 (CCK-8) was used to measure cell viability. Cell proliferation rate was determined by 5-acetyney-2 'deoxyuracil nucleoside (EdU). The apoptosis rate was determined by Hoechst 33258 staining. The protein expression levels of Cas-pase-3, Cyclin D1, nuclear transcription factor-κB p65 (NF-κB p65) and phosphorylation (p-)NF-κB p65 were determined by Western blotting.Results Compared with blank group, the expression levels of inflammatory cytokines IL-6, IL-8 and IL-1β were increased in LPS group (P<0.05), while the cell viability decreased (P<0.05). Compared with LPS group, the expression levels of inflammatory cyto-kines IL-6, IL-8 and IL-1β in LPS+4 and LPS+8 groups were decreased [LPS group: (40.24±4.59) ng/L, (249.53±8.60) ng/L, (85.22±2.64) ng/L; LPS+4 group: (23.23±2.86) ng/L, (170.99±10.62) ng/L, (25.87±2.91) ng/L; LPS+8 group: (20.38±2.57) ng/L, (127.57±5.49)ng/L, (27.56±2.89) ng/L] (P<0.05), while the cell viability was increased [LPS+4 group: (57.77±4.24) %; LPS+8 group: (71.80±7.40) %] (P<0.05). In this study, 4 mg/L azithromycin group with significant difference and lower concentration was selected as azithromycin group based on LPS group. The NF-κB pathway validation experiment was conducted by adding BAY 11-7082, an inhibitor of NF-κB signaling pathway, and Prostratin, a pathway activator, respectively. Compared with blank group, the cell proliferation rate and the ex-pression level of CyclinD1 protein in LPS group were decreased, while the apoptosis rate, caspase-3 and p-NF-κB p65 protein levels in-creased (P<0.05). The changes of the above indexes were significantly reversed in the azithromycin group and the inhibitor group (P< 0.05). Compared with azithromycin group, while cell proliferation rate and CyclinD1 protein expression level in azithromycin + inhibi-tor group were further increased, cell apoptosis rate, caspase-3 and p-NF-κB p65 protein expression levels were further decreased.Azithromycin + activator group significantly reversed the changes of the above indexes (P<0.05).Conclusion Azithromycin can pro-tect A549 cells from inflammatory damage by inhibiting NF-κB signaling pathway, promoting cell proliferation and inhibiting apoptosis. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
