MECHANISMS OF TETRANITROMETHANE TOXICITY AND CARCINOGENICITY

四硝基甲烷毒性和致癌机制

• 批准号: 6289888
• 负责人: DANIEL MORGAN
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6289888
• 关键词: SDS polyacrylamide gel electrophoresis; capillary electrophoresis; carcinogen testing; chemical carcinogen; chemical carcinogenesis; environmental toxicology; high performance liquid chromatography; membrane proteins; mutagen testing; nitroalkane; phosphorylation; respiratory neoplasm; respiratory toxin; toxin metabolism; tyrosine

The aim of this project is to investigate the mechanism(s) of tetranitromethane (TNM) pulmonary toxicity and carcinogenicity. TNM is commonly used in protein chemistry because of its specificity for nitrating the ortho position of tyrosine (Tyr) residues in proteins. Because of this highly specific reaction of TNM studies were conducted to evaluate the ability of TNM to nitrate cell membrane Tyr residues and to investigate the subsequent effects of Tyr nitration on cellular functions dependent upon Tyr kinase pathways. One such cellular function is the lipopolysaccharide/interferon (LPS/IFN) stimulated production of nitric oxide (NO) by macrophages. Rat alveolar macrophages (AM) were treated with 0, 0.312, 0.625, or 1.25 M TNM in serum-free media for 2 hours, then stimulated with LPS/IFN for 24 hours, and NO production measured. TNM treatment caused a dose-related decrease in the amount of NO released by stimulated AMs. Cell viability was not affected by these doses of TNM. Direct measurement of Tyr nitration using antibodies to N-tyr revealed an increased staining of AM membranes after exposure to TNM. These data suggest that TNM toxicity may be due in part to inhibition of Tyr kinase pathways caused by nitration of membrane Tyr. - tetranitromethane, pulmonary, nitrotyrosine, alveolar macrophage, nitric oxide, in vitro

该项目的目的是研究四硝基甲烷（TNM）肺毒性和致癌性的机制。 TNM 因其对蛋白质中酪氨酸 (Tyr) 残基邻位硝化的特异性而常用于蛋白质化学。由于 TNM 的这种高度特异性反应，进行了研究来评估 TNM 硝化细胞膜 Tyr 残基的能力，并研究 Tyr 硝化对依赖于 Tyr 激酶途径的细胞功能的后续影响。其中一种细胞功能是脂多糖/干扰素 (LPS/IFN) 刺激巨噬细胞产生一氧化氮 (NO)。将大鼠肺泡巨噬细胞 (AM) 用无血清培养基中的 0、0.312、0.625 或 1.25 M TNM 处理 2 小时，然后用 LPS/IFN 刺激 24 小时，并测量 NO 产量。 TNM 治疗导致受刺激的 AM 释放的 NO 量出现剂量相关的减少。细胞活力不受这些 TNM 剂量的影响。使用 N-tyr 抗体直接测量 Tyr 硝化显示，暴露于 TNM 后 AM 膜的染色增加。这些数据表明，TNM 毒性可能部分归因于膜 Tyr 硝化对 Tyr 激酶途径的抑制。 - 四硝基甲烷，肺，硝基酪氨酸，肺泡巨噬细胞，一氧化氮，体外