Early Inflammatory Response to Asbestos Exposure in Rat and Hamster Lungs: Role of Inducible Nitric Oxide Synthase

doi:10.1006/taap.2002.9388

Toxicology and Applied Pharmacology

Volume 181, Issue 2, 1 June 2002, Pages 93-105

https://doi.org/10.1006/taap.2002.9388 Get rights and content

Abstract

Recent studies have suggested that inducible nitric oxide synthase (iNOS) plays a role in the development of asbestos-related pulmonary disorders. The pulmonary reactions of rats and hamsters upon exposure to asbestos fibers are well known to be disparate. In addition, in vitro experiments have indicated that mononuclear phagocytes from hamsters, in contrast to those from rats, lack the iNOS pathway. Therefore, the purpose of this study was to investigate whether rats and hamsters differ in lung iNOS expression in vivo upon exposure to asbestos fibers and whether differences in iNOS induction are associated with differences in the acute pulmonary inflammatory reaction. Body weight, alveolar–arterial oxygen difference, differential cell count in bronchoalveolar lavage fluid, total protein leakage, lung myeloperoxidase activity and lipidperoxidation, wet/dry ratio, iNOS mRNA and protein expression, and nitrotyrosine staining of lung tissue were determined 1 and 7 days after intratracheal instillation of asbestos fibers in CD rats and Syrian golden hamsters. Exposure of rats to asbestos fibers resulted in enhanced pulmonary iNOS expression and nitrotyrosine staining together with an acute inflammation that was characterized by an influx of neutrophils, enhanced myeloperoxidase activity and lipid peroxidation, damage of the alveolar–capillary membrane, edema formation, and impairment of gas exchange. In comparison, instillation of asbestos fibers in hamsters resulted in a significantly milder inflammatory reaction of the lung with no induction of iNOS in pulmonary cells. The data obtained provide important information to understand the underlying mechanisms of species differences in the pulmonary response upon exposure to asbestos fibers.

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Citation Excerpt :
Several forms of lung diseases due to asbestos fibers' inhalation continue to be a significant problem globally and to challenge health care providers [22–24]. Although there are conflicting data, most published studies suggest that the NO production induced by inhalation of asbestos fibers in sufficient concentrations plays a dominant role in lung and pleura damage [25–51]. It has been supported that FeNO levels were increased in asbestos-exposed patients with borderline parenchymal changes on high resolution computed tomography [24].
Nitric oxide (NO) regulates various physiological and pathophysiological functions in the lungs. However, there is much less information about the effects of NO in the pleura. The present review aimed to explore the available evidence regarding the role of NO in pleural disease. NO, has a double-edged role in the pleural cavity. It is an essential signaling molecule mediating various physiological cell functions such as lymphatic drainage of the serous cavities, the immune response to intracellular multiplication of pathogens, and downregulation of neutrophil migration, but also induces genocytotoxic and mutagenic effects when present in excess. NO is implicated in the pathogenesis of asbestos-related or exudative pleural disease and mesothelioma. From a clinical point of view, the fraction of exhaled NO has been suggested as a potential non-invasive tool for the diagnosis of benign asbestos-related disorders. Under experimental conditions, NO-mimetics were found to attenuate hypoxia-induced therapy resistance in mesothelioma. Similarly, hybrid agents consisting of an NO donor coupled with a parent anti-inflammatory drug showed an enhancement of the anti-inflammatory activity of anti-inflammatory drugs. However, given the paucity of research work performed over the last years in this area, further research should be undertaken to establish reliable conclusions with respect to the feasibility of determining or targeting the NO signaling pathway for pleural disease diagnosis and therapeutic management.
Allele-specific effects of ecSOD on asbestos-induced fibroproliferative lung disease in mice
2011, Free Radical Biology and Medicine
Previous work by others suggests that there is a strain-dependent variation in the susceptibility to inflammatory lung injury in mice. Specifically, the 129/J mice appear to be more resistant to asbestos-induced pulmonary fibrosis than the C57BL/6 strain. A separate line of evidence suggests that extracellular superoxide dismutase (ecSOD) may play an important role in protecting the lung from such injuries. We have recently reported that the 129/J strain of mice has an ecSOD genotype and phenotype distinctly different from those of the C57BL/6 mice. In order to identify ecSOD as a potential “asbestos-injury resistance” gene, we bred congenic mice, on the C57BL/6 background, carrying the wild type (sod3^wt) or the 129/J (sod3¹²⁹) allele for ecSOD. This allowed us to examine the role of ecSOD polymorphism in susceptibility to lung injury in an otherwise identical genetic background. Interestingly, asbestos treatment induces a significant (~ 40%) increase in plasma ecSOD activity in the sod3¹²⁹ mice, but not in the sod3^wt mice. Asbestos administration results in a loss of ecSOD activity and protein from lung tissue of both congenic strains, but the lung ecSOD activity remains significantly higher in sod3¹²⁹ mice. As expected, asbestos treatment results in a significant recovery of ecSOD protein in bronchoalveolar lavage fluid (BALF). The BALF of sod3¹²⁹ mice also have significantly lower levels of proteins and inflammatory cells, especially neutrophils, accompanied by a significantly lower extent of lung injury, as measured by a pathology index score or hydroxyproline content. Immunohistochemistry reveals a significant loss of ecSOD from the tips of the respiratory epithelial cells in response to asbestos treatment and that the loss of immunodetectable ecSOD is compensated for by enzyme expression by infiltrating cells, especially in the sod3^wt mice. Our studies thus identify ecSOD as an important anti-inflammatory gene, responsible for most, if not all of the resistance to asbestos-induced lung injury reported for the 129/J strain of mice. The data further suggest allele-specific differences in the regulation of ecSOD expression. These congenic mice therefore represent a very useful model to study the role of this enzyme in all inflammatory diseases. Polymorphisms in human ecSOD have also been reported and it appears logical to assume that such variations may have a profound effect on disease susceptibility.
Extracellular superoxide dismutase inhibits inflammation by preventing oxidative fragmentation of hyaluronan
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Extracellular superoxide dismutase (EC-SOD) is expressed at high levels in lungs. EC-SOD has a polycationic matrix-binding domain that binds to polyanionic constituents in the matrix. Previous studies indicate that EC-SOD protects the lung in both bleomycin- and asbestos-induced models of pulmonary fibrosis. Although the mechanism of EC-SOD protection is not fully understood, these studies indicate that EC-SOD plays an important role in regulating inflammatory responses to pulmonary injury. Hyaluronan is a polyanionic high molecular mass polysaccharide found in the extracellular matrix that is sensitive to oxidant-mediated fragmentation. Recent studies found that elevated levels of low molecular mass hyaluronan are associated with inflammatory conditions. We hypothesize that EC-SOD may inhibit pulmonary inflammation in part by preventing superoxide-mediated fragmentation of hyaluronan to low molecular mass fragments. We found that EC-SOD directly binds to hyaluronan and significantly inhibits oxidant-induced degradation of this glycosaminoglycan. In vitro human polymorphic neutrophil chemotaxis studies indicate that oxidative fragmentation of hyaluronan results in polymorphic neutrophil chemotaxis and that EC-SOD can completely prevent this response. Intratracheal injection of crocidolite asbestos in mice leads to pulmonary inflammation and injury that is enhanced in EC-SOD knock-out mice. Notably, hyaluronan levels are increased in the bronchoalveolar lavage fluid after asbestos-induced pulmonary injury, and this response is markedly enhanced in EC-SOD knock-out mice. These data indicate that inhibition of oxidative hyaluronan fragmentation probably represents one mechanism by which EC-SOD inhibits inflammation in response to lung injury.
Molecular analysis of a multistep lung cancer model induced by chronic inflammation reveals epigenetic regulation of p16 and activation of the DNA damage response pathway
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The molecular hallmarks of inflammation-mediated lung carcinogenesis have not been fully clarified, mainly due to the scarcity of appropriate animal models. We have used a silica-induced multistep lung carcinogenesis model driven by chronic inflammation to study the evolution of molecular markers, genetic alterations. We analyzed markers of DNA damage response (DDR), proliferative stress, telomeric stress: δ-H2AX, p16, p53, TERT. Lung cancer-related epigenetic, genetic alterations, including promoter hypermethylation status of p16(CDKN2A), APC, CDH13, Rassf1, Nore1A, as well as mutations of Tp53, epidermal growth factor receptor, K-ras, N-ras, c-H-ras, have been also studied. Our results showed DDR pathway activation in preneoplastic lesions, in association with inducible nitric oxide synthase, p53 induction. p16 was also induced in early tumorigenic progression, was inactivated in bronchiolar dysplasias, tumors. Remarkably, lack of mutations of Ras, epidermal growth factor receptor, a very low frequency of Tp53 mutations suggest that they are not required for tumorigenesis in this model. In contrast, epigenetic alterations in p16(CDKN2A), CDH13, APC, but not in Rassf1, Nore1A, were clearly observed. These data suggest the existence of a specific molecular signature of inflammation-driven lung carcinogenesis that shares some, but not all, of the molecular landmarks of chemically induced lung cancer.
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¹: To whom correspondence should be addressed at Institut für Chirurgische Forschung-GroBhadern Klinikum der Universität München Marchioninistr, 27 D-81366 München, Germany. Fax: +49-89-7095-8897. E-mail: [email protected].

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Regular ArticleEarly Inflammatory Response to Asbestos Exposure in Rat and Hamster Lungs: Role of Inducible Nitric Oxide Synthase

Abstract

Environ. Res.

Arch. Biochem. Biophys.

Toxicol. Lett.

Ann. Occup. Hyg.

Biochim. Biophys. Acta

Toxicol. Lett.

Gene

Biochem. Biophys. Res. Commun.

Toxicol. Lett.

Cytokine

Anal. Biochem.

Arch. Biochem. Biophys.

Free Radical Biol. Med.

Anal. Biochem.

Nitric Oxide

Atherosclerosis

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Intratracheal instillation of silica up-regulates inducible nitric oxide synthase gene expression and increases nitric oxide production in alveolar macrophages and neutrophils

Am. J. Respir. Cell Mol. Biol.

Comparison of inducible nitric oxide synthase gene expression and lung inflammation following intratracheal instillation of silica, coal, carbonyl iron, or titanium dioxide in rats

J. Toxicol. Environ. Health

Tumours of the respiratory tract in rats and hamsters following chronic inhalation of engine exhaust emissions

J. Appl. Toxicol.

Species variation in lung antioxidant enzyme activities

J. Appl. Physiol.

The role of inflammation, oxidative stress, and proliferation in silica-induced lung disease: A species comparison

J. Environ. Pathol. Toxicol. Oncol.

Multiple genetic alterations in hamster pancreatic ductal adenocarcinomas

Cancer Res.

Transoral tracheal intubation of rodents using a fiberoptic laryngoscope

Lab. Anim. Sci.

Nitric oxide participates in early events associated with NNMU-induced acute lung injury in rats

Am. J. Physiol.

Species differences in NO formation by rat and hamster alveolar macrophages in vitro

Am. J. Respir. Cell Mol. Biol.

Alveolar macrophage cytokine and growth factor production in a rat model of crocidolite-induced pulmonary inflammation and fibrosis

J. Toxicol. Environ. Health

Formation of nitric oxide-derived inflammatory oxidants by myeloperoxidase in neutrophils

Nature

Comparison of pulmonary and pleural responses of rats and hamsters to inhaled refractory ceramic fibers

Toxicol. Sci.

Interstitial pulmonary fibrosis induced in hamsters by intratracheally administered chrysotile asbestos

Am. Rev. Respir. Dis.

Cytotoxicity of long and short crocidolite asbestos fibers in vitro and in vivo

Cancer Res.

Inhibition of alveolar type II cell ATP and surfactant synthesis by nitric oxide

Am. J. Physiol.

Nitration of surfactant protein A results in decreased ability to aggregate lipids

Am. J. Physiol.

Regular Article
Early Inflammatory Response to Asbestos Exposure in Rat and Hamster Lungs: Role of Inducible Nitric Oxide Synthase