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目的:观察重组高迁移率族蛋白B1(high mobility group box 1,HMGB1)对肺上皮细胞致炎细胞因子释放的影响,并初步探讨其机制。方法:采用BEAS-2B人正常肺上皮细胞,观察1、10、100、1 000μg/L重组HMGB1蛋白对细胞培养上清液中致炎细胞因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素(interleukin,IL)-1β、IL-6含量的影响,及10 mg/L Toll样受体4(Toll-like receptor 4,TLR4)抗体处理对HMGB1诱导致炎细胞因子释放的抑制作用。TNF-α、IL-1β和IL-6含量均采用酶联免疫吸附试验检测。结果:不同剂量重组HMGB1诱导肺上皮细胞12 h后,培养上清液中TNF-α、IL-1β、IL-6含量均呈HMGB1剂量依赖性升高;100μg/L HMGB1分别诱导肺上皮细胞3、6、12和24 h后,培养上清液中TNF-α、IL-1β、IL-6含量均呈显著性升高(P<0.05或P<0.01),IL-1β、IL-6含量随诱导时间延长而升高,而TNF-α含量于诱导后6 h达峰值;TLR4抗体处理后TNF-α、IL-1β、IL-6释放均受到部分抑制(P<0.05或P<0.01)。结论:HMGB1对肺上皮细胞释放TNF-α、IL-1β和IL-6具有诱导作用,其机制可能与胞膜TLR4有关。
OBJECTIVE: To investigate the effect of recombinant HMGB1 on the release of proinflammatory cytokines in lung epithelial cells and to explore its mechanism. Methods: BEAS-2B human normal lung epithelial cells were used to observe the effects of 1, 10, 100, 1 000 μg / L HMGB1 recombinant protein on the expression of tumor necrosis factor-α (TNF) α, interleukin (IL) -1β and IL-6, and treatment with 10 mg / L Toll-like receptor 4 (TLR4) antibody on HMGB1-induced proinflammatory cytokine release Inhibition. TNF-α, IL-1β and IL-6 levels were detected by enzyme-linked immunosorbent assay. Results: After induced with different doses of recombinant HMGB1 for 12 h, the levels of TNF-α, IL-1β and IL-6 in culture supernatants were increased in a dose-dependent manner. HMGB1 at 100 μg / L induced a significant increase in the number of lung epithelial cells 3 (P <0.05 or P <0.01). The contents of IL-1β and IL-6 in the culture supernatants were significantly increased at 6, 12 and 24 h (P <0.05 or P <0.01). The release of TNF-α, IL-1β and IL-6 were partially inhibited by TLR4 antibody treatment (P <0.05 or P <0.01) . CONCLUSION: HMGB1 induces the release of TNF-α, IL-1β and IL-6 from lung epithelial cells. The mechanism may be related to the membrane TLR4.