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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 effect of formaldehyde on human bronchial epithelial cells and the secretion of related inflammatory effector molecules. Methods In vitro exposure system of formaldehyde gas-liquid interface was established by fully confluent and well-differentiated human bronchial epithelial cells BEAS-2B and petri-PERM gas permeable culture medium. A control (sterile air) group was set up and a high dose (0.30 mg / (0.10 mg / m3) formaldehyde exposure group and low dose (0.10 mg / m3) formaldehyde exposure + N-acetylcysteine (NAC, 0.01 mol / L) protection group were cultured for 1 h and 8 h Cell proliferation toxicity, SOD activity, GSH, GSSG, NO, Interleukin 2 (IL-2) and intracellular nitric oxide synthase (NOS) activity, and observed cell morphology. Results The proliferation and SOD activity of BEAS-2B cells exposed to low and high doses of formaldehyde exposure group were higher than those of the control group at 1 h and 8 h after exposure. The cell proliferation toxicity and the activity of SOD in the low-dose formaldehyde exposure + NAC group were lower than those in the control group GSH / GSSG in BEAS-2B cells exposed to low and high dose formaldehyde exposure groups was lower than that of the control group, GSH / GSSG values in low dose formaldehyde exposure + NAC protection groups were higher than those in low dose formaldehyde exposure groups; low dose formaldehyde The concentrations of NO, IL-2 and NOS in the culture supernatant of BEAS-2B cells exposed group were higher than those in the control group. The concentrations of NO, IL-2 and NOS in the low-dose formaldehyde exposure + NAC group were lower than those in the low-dose formaldehyde exposure group All were statistically significant (P <0.05). Low-dose formaldehyde exposure + NAC protection group and the control group of the above indicators were no significant difference between. Compared with the control group, the morphology of the cells in the formaldehyde exposure group changed after 8 hours of exposure, while the low dose formaldehyde exposure + NAC protection group showed no obvious morphological changes. CONCLUSION: Formaldehyde can cause oxidative damage to bronchial epithelial cells, change intracellular redox status, and induce the bronchial epithelial cells to synthesize inflammatory mediators, which may be involved in the initiation of airway inflammation. Antioxidant NAC can inhibit the oxidation of bronchial epithelial cells exposed to low doses of formaldehyde Injury and release of inflammatory mediators.