甲醛气液界面暴露对人支气管上皮细胞的氧化损伤效应  

Oxidative damage of human bronchial cells induced by formaldehyde: an experimental study of air-liquid interface exposure in vitro

作　　者：管东波[1] 宋宏[2] 

机构地区：[1]佛山市南海区疾病预防控制中心职业卫生科,广东528200 [2]中山大学公共卫生学院

出　　处：《环境与健康杂志》2013年第9期759-762,F0003,共5页Journal of Environment and Health

基　　金：国家自然科学基金(30872080)

摘　　要：目的探讨甲醛对人支气管上皮细胞的氧化损伤效应及相关炎症效应分子的分泌水平。方法利用完全融合并分化完全的人支气管上皮细胞BEAS-2B及petri-PERM透气培养皿建立甲醛气液界面体外暴露系统,设立对照(无菌空气)组、高剂量(0.30 mg/m3)甲醛暴露组、低剂量(0.10 mg/m3)甲醛暴露组、低剂量(0.10 mg/m3)甲醛暴露+N-乙酰半胱氨酸(NAC,0.01 mol/L)保护组,分别培养1 h和8 h,测定细胞增殖毒性、超氧化物歧化酶(SOD)活力和细胞内还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)、细胞培养上清中一氧化氮(NO)、白介素2(IL-2)浓度以及细胞内一氧化氮合酶(NOS)活力,并观察细胞形态。结果染毒1 h和8 h后,低剂量和高剂量甲醛暴露组的BEAS-2B细胞增殖毒性和SOD活力高于对照组,低剂量甲醛暴露+NAC保护组细胞增殖毒性、SOD活力低于低剂量甲醛暴露组;低剂量和高剂量甲醛暴露组BEAS-2B细胞内GSH/GSSG值低于对照组,低剂量甲醛暴露+NAC保护组GSH/GSSG值高于低剂量甲醛暴露组;低剂量甲醛暴露组BEAS-2B细胞培养上清中NO、IL-2浓度和NOS活力高于对照组,低剂量甲醛暴露+NAC保护组NO、IL-2浓度和NOS活力低于低剂量甲醛暴露组,差异均有统计学意义(P<0.05)。低剂量甲醛暴露+NAC保护组与对照组的上述各指标间差异均无统计学意义。与对照组相比,甲醛暴露组染毒8 h后细胞形态有改变,而低剂量甲醛暴露+NAC保护组无明显形态学改变。结论甲醛能对支气管上皮细胞造成氧化损伤,改变细胞内氧化还原状态,并可能诱导支气管上皮细胞合成炎症介质参与气道炎症的启动;而抗氧化剂NAC可以抑制低剂量甲醛暴露对支气管上皮细胞的氧化损伤和炎症介质释放。Objective To investigate the oxidative damage effects of formaldehyde on human bronchial epithelial cells and the secretion of inflammatory effect molecules. Methods Formaldehyde exposure system in vitro was established with hologram and completely differentiated human bronchial epithelial ceils and petri-PERM culture dish. Four groups were set：control group（fihrated air）,high dose of formaldehyde group（0.30 mg/m3）, low dose of formaldehyde group （0.10 mg/m3） and low dose of formaldehyde exposure （0.10 mg/m3） ＋ N-acetylcystein （NAC, 0.01 mol/L） protection group, and each group had two time spots （1 h and 8 h）. After exposure, the contents of GSH and GSSG in cells,the activities of SOD and NOS,and the concentrations of NO and IL-2 in the culture media were detected, the change of cell morphology was observed. Results Significant morphological differences were found in the cells between the experimental and the control groups, especially in high dose and 8 h exposure group. Compared with the control group, the ratios of GSH and GSSG in BEAS-2B cells in the low dose and the high dose group were lower, the activities of SOD and NOS in cells and the concentrations of NO and IL-2 were significantly higher （P〈0.05, P〈0.01）. Compared with the low dose formaldehyde group,the ratio of GSH and GSSG in BEAS-2B cells in low dose formaldehyde plus N-acetylcystein group was higher, the activities of SOD and NOS in cells and the concentrations of NO and IL-2 were lower, the differences were significant （P〈0.05, P〈0.01）. Conclusion Formaldehyde may cause oxidative damage, change the intracellular redox state and induce the synthesis of the inflammatory factors in bronchial epithelial cells, which may be involved in the start of the airway inflammation, and N-acetylcystein may present antagonism to these adverse effects.

关 键 词：甲醛 气液界面 人支气管上皮细胞 氧化损伤 

分 类 号：R994.6[医药卫生—毒理学]