脂氧素A4对脂多糖诱导气道上皮细胞炎症反应的影响  

The effect of lipoxin A4 on lipopolysaccharide-induced inflammatory responses in airway epithelial cells

作　　者：吴镇杰 WU Zhenjie(Respiratory and Critical Care Medicine Department,Taizhou Hospital of Zhejiang Province,Taizhou 317000,Zhejiang,China)

机构地区：[1]浙江省台州医院呼吸与危重症医学科,浙江台州3170001

出　　处：《中国现代医生》2025年第6期40-44,共5页China Modern Doctor

摘　　要：目的研究脂氧素A4(lipoxin A4,LXA4)对脂多糖(lipopolysaccharide,LPS)诱导气道上皮细胞炎症反应的影响,并进一步探讨其作用机制。方法通过LPS刺激正常人支气管上皮细胞(normal human bronchial epithelial cells,BEAS-2B)构建体外炎症模型。将BEAS-2B细胞分为4组。对照组不做任何处理;LPS组:BEAS-2B细胞与100ng/ml LPS共同孵育24h;LXA4组:100nmol/L LXA4预处理30min;LPS+LXA4组:将BEAS-2B细胞在含100nmol/L LXA4的培养基中预处理30min,再与100ng/ml LPS共同孵育24h。通过细胞活性检测试验检测LXA4的细胞毒性,通过酶联免疫吸附试验检测细胞培养上清液中白细胞介素(interleukin,IL)-6、IL-8及肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的浓度,通过免疫印迹(Western blot)法检测细胞基质金属蛋白酶-9(matrix metallo proteinase-9,MMP-9)蛋白的表达。最后用Western blot法检测核转录因子κB(nuclear factor-κB,NF-κB)信号通路相关蛋白的表达。结果①在药物浓度<100nmol/L的情况下,LXA4对BEAS-2B细胞未产生明显的细胞毒性。用100ng/ml LPS刺激BEAS-2B细胞24h后,细胞培养上清液IL-6、IL-8和TNF-α的浓度与对照组相比显著增加(P<0.01)。100nmol/L的LXA4预处理30min可显著减少LPS诱导的BEAS-2B细胞产生IL-6、IL-8和TNF-α(P<0.05)。②与对照组比较,LPS组MMP-9蛋白的表达明显增加(P<0.01);与LPS组比较,LPS+LXA4组MMP-9蛋白的表达明显减少(P<0.05)。③随着LPS刺激时间的延长,上皮细胞胞浆p65的表达明显减少。同时,LPS刺激BEAS-2B细胞可诱发核转录因子抑制蛋白κB-α(inhibitor ofκB-α,IκB-α)降解,在刺激30min后作用最显著。而用100nmol/L LXA4预处理30min后,IκB-α与胞浆NF-κB p65的表达减少程度明显减弱(P<0.05)。结论LXA4抑制LPS诱导的气道上皮细胞炎症介质IL-6、IL-8和TNF-α的产生及MMP-9蛋白的表达,其作用机制可能是通过抑制NF-κB信号通路实现。Objective To investigate the impact of lipoxin A4(LXA4)on the inflammatory response in airway epithelial cells induced by lipopolysaccharide(LPS),and to further explore its underlying mechanism of action.Methods An in vitro inflammation model was established by stimulating normal human bronchial epithelial cells(BEAS-2B)with LPS.BEAS-2B cells were divided into four groups.Control group:No treatment was applied;LPS group:BEAS-2B cells were incubated with 100 ng/ml LPS for 24 hours.LXA4 group:BEAS-2B cells were pretreated with 100 nmol/L LXA4 for 30 minutes.LPS+LXA4 group:BEAS-2B cells were pretreated with 100 nmol/L LXA4 for 30 minutes in culture medium and then incubated with 100 ng/ml LPS for an additional 24 hours.Cell viability assays were conducted to assess the cytotoxicity of LXA4.Enzyme linked immunosorbent assay was used to measure the concentrations of interleukin(IL)-6,IL-8,and tumor necrosis factor-α(TNF-α)in the cell culture supernatants.Western blot analysis was employed to detect the expression of matrix metalloproteinase-9(MMP-9)protein in the cells.Finally,Western blot analysis was performed to examine the expression of proteins related to the nuclear factor-κB(NF-κB)signaling pathway.Results①LXA4 exhibited negligible cytotoxicity towards BEAS-2B cells at concentrations below 100 nmol/L.Relative to control group,the exposure of BEAS-2B cells to 100ng/ml of LPS for a period of 24 hours led to a substantial augmentation in the levels of IL-6,IL-8,and TNF-αwithin the cellular supernatants(P<0.01).Pretreatment with 100 nmol/L LXA4 for a period of 30 minutes markedly attenuated the levels of IL-6,IL-8,and TNF-αin LPS-stimulated BEAS-2B cells(P<0.05).②Compared with control group,the expression of MMP-9 protein was significantly increased in LPS group(P<0.01).When compared to LPS group,the expression of MMP-9 protein was significantly decreased in LPS+LXA4 group(P<0.05).③The expression of p65 within the cytoplasm of BEAS-2B cells exhibited a notable decline as the duration of LPS stimulation

关 键 词：脂氧素 脂多糖 气道上皮细胞 炎症 

分 类 号：R562[医药卫生—呼吸系统]