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共同培养的与潮湿建筑相关的微生物加州链霉菌和黑葡萄穗霉通过氧化应激诱导免疫毒性和遗传毒性反应。

Co-cultivated damp building related microbes Streptomyces californicus and Stachybotrys chartarum induce immunotoxic and genotoxic responses via oxidative stress.

作者信息

Markkanen Penttinen Piia, Pelkonen Jukka, Tapanainen Maija, Mäki-Paakkanen Jorma, Jalava Pasi I, Hirvonen Maija-Riitta

机构信息

Department of Environmental Health, National Public Health Institute, Kuopio, Finland.

出版信息

Inhal Toxicol. 2009 Aug;21(10):857-67. doi: 10.1080/08958370802526873.

DOI:10.1080/08958370802526873
PMID:19459771
Abstract

Oxidative stress has been proposed to be one mechanism behind the adverse health outcomes associated with living in a damp indoor environment. In the present study, the capability of damp building-related microbes Streptomyces californicus and Stachybotrys chartarum to induce oxidative stress was evaluated in vitro. In addition, the role of oxidative stress in provoking the detected cytotoxic, genotoxic, and inflammatory responses was studied by inhibiting the production of reactive oxygen species (ROS) using N-acetyl-l-cysteine (NAC). RAW264.7 macrophages were exposed in a dose- and time-dependent manner to the spores of co-cultivated S. californicus and S. chartarum, to their separately cultivated spore-mixture, or to the spores of these microbes alone. The intracellular peroxide production and cytotoxicity were measured by flow cytometric analysis, nitric oxide production was analyzed by the Griess method, DNA damage was determined by the comet assay, and cytokine production was measured by an immunochemical ELISA (enzyme-linked immunosorbent assay). All the studied microbial exposures triggered oxidative stress and subsequent cellular damage in RAW264.7 macrophages. The ROS scavenger, NAC, prevented growth arrest, apoptosis, DNA damage, and cytokine production induced by the co-culture since it reduced the intracellular level of ROS within macrophages. In contrast, the DNA damage and cell cycle arrest induced by the spores of S. californicus alone could not be prevented by NAC. Bioaerosol-induced oxidative stress in macrophages may be an important mechanism behind the frequent respiratory symptoms and diseases suffered by residents of moisture damaged buildings. Furthermore, microbial interactions during co-cultivation stimulate the production of highly toxic compound(s) which may significantly increase oxidative damage.

摘要

氧化应激被认为是与居住在潮湿室内环境相关的不良健康后果背后的一种机制。在本研究中,对与潮湿建筑相关的微生物加州链霉菌和黑曲霉诱导氧化应激的能力进行了体外评估。此外,通过使用N-乙酰-L-半胱氨酸(NAC)抑制活性氧(ROS)的产生,研究了氧化应激在引发检测到的细胞毒性、遗传毒性和炎症反应中的作用。RAW264.7巨噬细胞以剂量和时间依赖性方式暴露于共培养的加州链霉菌和黑曲霉的孢子、它们单独培养的孢子混合物或这些微生物的单独孢子中。通过流式细胞术分析测量细胞内过氧化物的产生和细胞毒性,通过Griess法分析一氧化氮的产生,通过彗星试验确定DNA损伤,并通过免疫化学ELISA(酶联免疫吸附测定)测量细胞因子的产生。所有研究的微生物暴露均引发了RAW264.7巨噬细胞中的氧化应激和随后的细胞损伤。ROS清除剂NAC可防止共培养诱导的生长停滞、细胞凋亡、DNA损伤和细胞因子产生,因为它降低了巨噬细胞内ROS的水平。相比之下,NAC不能预防单独的加州链霉菌孢子诱导的DNA损伤和细胞周期停滞。生物气溶胶诱导的巨噬细胞氧化应激可能是潮湿受损建筑物居民频繁出现呼吸道症状和疾病背后的重要机制。此外,共培养期间的微生物相互作用会刺激高毒性化合物的产生,这可能会显著增加氧化损伤。

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