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过氧化氢可诱导呼吸道上皮细胞中的DNA单链断裂。

Hydrogen peroxide induces DNA single strand breaks in respiratory epithelial cells.

作者信息

McDonald R J, Pan L C, St George J A, Hyde D M, Ducore J M

机构信息

Department of Pediatrics, University of California, Davis.

出版信息

Inflammation. 1993 Dec;17(6):715-22. doi: 10.1007/BF00920476.

Abstract

The respiratory epithelium is often exposed to oxidant gases, including ozone from photochemical smog and toxic oxygen metabolites released from neutrophils recruited in conditions of airway inflammation. We evaluated DNA single strand break formation by alkaline elution as a measure of oxidant-induced DNA damage to bronchial epithelial cells. Human AdenoSV-40-transformed bronchial epithelial cells (BEAS), subclone R1.4 or nonhuman primate bronchial epithelial cells were cultured in growth factor supplemented Ham's F12 medium on polycarbonate filters. DNA was labeled by incubation with [3H]thymidine. Cells were incubated for 1 h in HBSS or HBSS and increasing concentrations of hydrogen peroxide (H2O2). Cells incubated in H2O2 demonstrated dose-dependent increases in strand break formation, and BEAS cells were more sensitive to H2O2-induced injury than primary bronchial epithelial cells. The addition of catalase or preincubation of cells with the iron chelator desferoxamine prevented H2O2-induced strand breakage. DNA strand break formation may be an important mechanism of oxidant injury in respiratory epithelial cells.

摘要

呼吸道上皮经常暴露于氧化性气体中,包括光化学烟雾中的臭氧以及气道炎症时募集的中性粒细胞释放的有毒氧代谢产物。我们通过碱性洗脱评估DNA单链断裂的形成,以此作为氧化性物质诱导支气管上皮细胞DNA损伤的一种测量方法。人腺病毒SV - 40转化的支气管上皮细胞(BEAS)、亚克隆R1.4或非人灵长类支气管上皮细胞在补充了生长因子的Ham's F12培养基中于聚碳酸酯滤膜上培养。通过与[3H]胸腺嘧啶孵育来标记DNA。将细胞在HBSS或HBSS以及浓度递增的过氧化氢(H2O2)中孵育1小时。在H2O2中孵育的细胞显示出链断裂形成呈剂量依赖性增加,并且BEAS细胞比原代支气管上皮细胞对H2O2诱导的损伤更敏感。加入过氧化氢酶或用铁螯合剂去铁胺对细胞进行预孵育可防止H2O2诱导的链断裂。DNA链断裂的形成可能是呼吸道上皮细胞氧化性损伤的一个重要机制。

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