【摘要】 目的探讨炭黑颗粒对人支气管上皮细胞(16HBE)自噬水平及炎症反应的影响。方法体外培养16HBE细胞,分为0μg/ml(空白对照组)及12.5、25、50、100μg/ml炭黑颗粒染毒组,染毒时间24 h。采用CCK-8法检测细胞存活率,Western blot法测定细胞内自噬体蛋白LC3-Ⅱ的表达水平,酶联免疫吸附(ELISA)法检测细胞上清液中细胞因子白介素8(IL-8)、白介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)和白介素6(IL-6)的含量。结果人支气管上皮细胞16HBE经12.5、25、50、100μg/ml炭黑颗粒悬液染毒24 h后,细胞存活率均低于空白对照组,且随浓度增加细胞存活率逐渐下降,呈明显的剂量-反应关系;各剂量染毒组16HBE细胞的自噬体蛋白LC3-Ⅱ表达水平均升高,其中100μg/ml组LC3-Ⅱ蛋白表达高于空白对照组,差异有统计学意义(P<0.05);25、50、100μg/ml染毒组的细胞上清液中IL-8含量高于空白对照组,100μg/ml染毒组的IL-1β含量高于对照组,差异有统计学意义(P<0.05)。结论炭黑颗粒可导致人支气管上皮细胞内自噬体蛋白LC3-Ⅱ表达水平增加,诱导细胞释放高水平的IL-8、IL-1β等炎性细胞因子。更多还原

【Abstract】 Objective To explore the effects of carbon black particles on autophagy and inflammatory responses in human bronchial epithelial cells(16 HBE). Methods The 16 HBE cells were treated with carbon black particles at the doses of 0, 12.5,25, 50 and 100 μg/ml for 24 h. Cell viability, LC3-Ⅱ protein level in cells and the secretions of IL-8, IL-1β, TNF-α and IL-6 in supernatants were assessed by CCK-8 kit, Western blot and ELISA, respectively. Results The cell viability was significantly decreased induced by carbon black particles in a dose-response manner(P<0.05). The expression of LC3-Ⅱ protein in 16 HBE cells was increased in all carbon black particles-treated goups, but the LC3-Ⅱ protein level in the group treated with 100 μg/ml carbon black particles was significantly higher than that in control group(P <0.05). Compared with control group, the IL-8 expression was significantly increased in groups treated by carbon black particles at 25 μg/ml or higher, and the IL-1β level was also significantly elevated in group treated by 100 μg/ml carbon black particles(all P<0.05). Conclusion Carbon black particles may increase LC3-Ⅱ protein expression and induce IL-8 and IL-1β secretion in 16 HBE cells.更多还原