Hardwick S J, Adam A, Smith L L, Cohen G M
Department of Pharmacology, University of London, UK.
Environ Health Perspect. 1990 Apr;85:129-33. doi: 10.1289/ehp.85-1568319.
In order to facilitate the study of oxidative stress in lung tissue, rat lung slices with impaired antioxidant defenses were prepared and used. Incubation of lung slices with the antineoplastic agent 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) (100 microM) in an amino acid-rich medium for 45 min produced a near-maximal (approximately 85%), irreversible inhibition of glutathione reductase, accompanied by only a modest (approximately 15%) decrease in pulmonary nonprotein sulfhydryls (NPSH) and no alteration in intracellular ATP, NADP+, and NADPH levels. The amounts of NADP(H), ATP, and NPSH were stable over a 4-hr incubation period following the removal from BCNU. The viability of the system was further evaluated by measuring the rate of evolution of 14CO2 from D-[14C(U)]-glucose. The rates of evolution were almost identical in the compromised system when compared with control slices over a 4-hr time period. By using slices with compromised oxidative defenses, preliminary results have been obtained with paraquat, nitrofurantoin, and 2,3-dimethoxy-1,4-naphthoquinone.
为便于研究肺组织中的氧化应激，制备并使用了抗氧化防御受损的大鼠肺切片。在富含氨基酸的培养基中，将抗肿瘤药物1,3-双(2-氯乙基)-1-亚硝基脲(BCNU)(100微摩尔)与肺切片孵育45分钟，可产生近乎最大程度(约85%)的、不可逆的谷胱甘肽还原酶抑制作用，同时肺非蛋白巯基(NPSH)仅适度降低(约15%)，细胞内ATP、NADP⁺和NADPH水平无变化。从BCNU中取出后，在4小时的孵育期内，NADP(H)、ATP和NPSH的量保持稳定。通过测量D-[14C(U)]-葡萄糖产生14CO2的速率，进一步评估了该系统的活力。在4小时的时间段内，与对照切片相比，受损系统中的产生速率几乎相同。通过使用氧化防御受损的切片，已获得了百草枯、呋喃妥因和2,3-二甲氧基-1,4-萘醌的初步结果。
