Minakami H, Arai H, Nakano M, Sugioka K, Suzuki S, Sotomatsu A
Department of Obstetrics and Gynecology, Jichi Medical School, Tochigi, Japan.
Biochem Biophys Res Commun. 1988 Jun 30;153(3):973-8. doi: 10.1016/s0006-291x(88)81323-8.
In order to evaluate the O-2 participation in NADPH-dependent microsomal lipid peroxidation, we used reconstructed system which contained detergent-solubilized NADPH-dependent cytochrome P-450 reductase, cytochrome P-450, phospholipid liposomes, NADPH and Fe3+-ADP. Lipid peroxidation, monitored by the formation of thiobarbituric acid-reactive substance, was increased with increasing concentration of detergent-solubilized NADPH cytochrome P-450 reductase, cytochrome P-450 or Fe3+-ADP. Cytochrome P-450-dependent lipid peroxidation was parallel to O-2 generation monitored by chemiluminescence probe with 2-methyl-6-(p-methoxyphenol)-3,7-dihydroimidazo[1,2-a]pyrazin++ +-3-one. Lipid peroxidation was significantly inhibited by superoxide dismutase, but not by catalase or sodium benzoate. The reconstructed system herein described is considered to be very close to NADPH-dependent microsomal lipid peroxidation system.
为了评估O₂参与NADPH依赖性微粒体脂质过氧化反应的情况，我们使用了一个重构系统，该系统包含去污剂溶解的NADPH依赖性细胞色素P-450还原酶、细胞色素P-450、磷脂脂质体、NADPH和Fe³⁺-ADP。通过硫代巴比妥酸反应性物质的形成来监测脂质过氧化反应，随着去污剂溶解的NADPH细胞色素P-450还原酶、细胞色素P-450或Fe³⁺-ADP浓度的增加，脂质过氧化反应增强。细胞色素P-450依赖性脂质过氧化反应与通过化学发光探针2-甲基-6-(对甲氧基苯酚)-3,7-二氢咪唑并[1,2-a]吡嗪-3-酮监测的O₂生成平行。脂质过氧化反应受到超氧化物歧化酶的显著抑制，但不受过氧化氢酶或苯甲酸钠的抑制。本文所述的重构系统被认为与NADPH依赖性微粒体脂质过氧化系统非常接近。
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