Pipe R K
Natural Environment Research Council, Plymouth Marine Laboratory, Citadel Hill, Plymouth, UK.
Dev Comp Immunol. 1992 Mar-Jun;16(2-3):111-22. doi: 10.1016/0145-305x(92)90012-2.
Generation of superoxide anion by stimulated haemocytes of Mytilus edulis was demonstrated using dihydrorhodamine 123 and quantified using reduction of nitroblue tetrazolium (NBT). In the presence of zymosan or phorbol myristate acetate, there was an increased reduction of NBT to formazan. The addition of superoxide dismutase (SOD) and iodoacetamide to the incubation medium resulted in a significant reduction in deposition of reduced formazan. Incubation of haemocytes with the SOD inhibitor diethyldithiocarbamate (DDC) gave rise to a small but significant increase in NBT reduction. The production of hydrogen peroxide by haemocytes was quantified using horseradish peroxidase-dependent oxidation of phenol red. The presence of SOD in the incubation medium together with zymosan resulted in a significant increase in H2O2 production. Haemocytes incubated with DDC prior to the assay or with sodium nitroprusside during the assay showed a decrease in H2O2 production with increasing concentration of the inhibitor.
利用二氢罗丹明123证明了紫贻贝受刺激血细胞中超氧阴离子的产生，并通过硝基蓝四唑（NBT）的还原进行定量。在存在酵母聚糖或佛波酯肉豆蔻酸酯的情况下，NBT还原为甲臜的程度增加。向孵育培养基中添加超氧化物歧化酶（SOD）和碘乙酰胺导致还原甲臜沉积显著减少。用SOD抑制剂二乙基二硫代氨基甲酸盐（DDC）孵育血细胞导致NBT还原有小幅但显著的增加。利用辣根过氧化物酶依赖的酚红氧化对血细胞产生过氧化氢进行定量。孵育培养基中SOD与酵母聚糖共同存在导致H2O2产生显著增加。在测定前用DDC孵育血细胞或在测定期间用硝普钠孵育血细胞，随着抑制剂浓度增加，H2O2产生减少。
