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暴露于可吸入性α-石英的人肺上皮细胞中的细胞毒性和DNA损伤。

Cytotoxicity and DNA-damage in human lung epithelial cells exposed to respirable alpha-quartz.

作者信息

Fanizza Carla, Ursini Cinzia Lucia, Paba Emilia, Ciervo Aureliano, Di Francesco Arianna, Maiello Raffaele, De Simone Paolo, Cavallo Delia

机构信息

Department of Occupational Hygiene, ISPESL, National Institute for Occupational Safety and Prevention, Via Fontana Candida 1, 00040 Monteporzio Catone, Rome, Italy.

出版信息

Toxicol In Vitro. 2007 Jun;21(4):586-94. doi: 10.1016/j.tiv.2006.12.002. Epub 2006 Dec 16.

Abstract

Occupational exposure to respirable crystalline silica is associated with the development of silicosis, lung cancer and airways diseases. In order to assess cytotoxic effects and direct-oxidative DNA damage induced by short-term exposure to different doses of respirable alpha-quartz (NIST SRM1878a), we conducted a study using A549 cells. The cells were exposed to alpha-quartz at 25, 50, 100 microg/ml for 4 h and analysed by scanning electron microscope (SEM) and LDH release assay for cytotoxic effect evaluation. Cells were also exposed to 10, 25, 50, 100 microg/ml of alpha-quartz for 2 h and 4 h and analysed by Fpg comet test to evaluate direct and oxidative DNA damage. SEM observations of treated cells showed bleb development at lower doses and alterations of microvilli morphology at the highest dose. A slight LDH release was found only at 100 microg/ml. Fpg comet test showed a dose-related oxidative DNA damage in cells exposed for 2 h to quartz. Cells exposed for 4h at the same concentrations showed a dose-related direct DNA damage and the presence of oxidative DNA damage at lower doses. The bleb induction on cell surface evidenced by SEM at lower doses correlates with the presence of oxidative DNA damage at 4 h. The cell surface modifications observed by SEM at 100 microg/ml indicate that high doses of quartz induce more evident cytotoxic effects confirmed by LDH analysis and correlate with the genotoxicity showed by comet assay.

摘要

职业性接触可吸入结晶二氧化硅与矽肺、肺癌和气道疾病的发生有关。为了评估短期接触不同剂量的可吸入α-石英(NIST SRM1878a)所诱导的细胞毒性作用和直接氧化性DNA损伤,我们使用A549细胞进行了一项研究。将细胞暴露于浓度为25、50、100微克/毫升的α-石英中4小时,并通过扫描电子显微镜(SEM)和乳酸脱氢酶(LDH)释放试验进行分析,以评估细胞毒性作用。细胞还分别暴露于浓度为10、25、50、100微克/毫升的α-石英中2小时和4小时,并通过Fpg彗星试验进行分析,以评估直接和氧化性DNA损伤。对处理过的细胞进行SEM观察发现,较低剂量时出现了小泡形成,最高剂量时微绒毛形态发生了改变。仅在100微克/毫升时发现有轻微的LDH释放。Fpg彗星试验显示,暴露于石英2小时的细胞中存在剂量相关的氧化性DNA损伤。在相同浓度下暴露4小时的细胞显示出剂量相关的直接DNA损伤,且在较低剂量时存在氧化性DNA损伤。较低剂量下SEM所证实的细胞表面小泡诱导与4小时时氧化性DNA损伤的存在相关。在100微克/毫升时SEM观察到的细胞表面修饰表明,高剂量石英诱导了更明显的细胞毒性作用,这通过LDH分析得到证实,并与彗星试验所显示的遗传毒性相关。

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