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鸡肝微粒体细胞色素P450酶系检测方法的建立
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杨海峰,江善祥
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（南京农业大学 动物医学院， 江苏 南京210095）
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摘要：
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为建立鸡肝微粒体细胞色素P450(CYP450)酶系活性的测定方法，应用超速离心法提取了12只60日龄依莎公鸡的肝微粒体，采用Bradford法测定肝微粒体蛋白含量，紫外分光光度法测定CYP450酶系活性。结果显示，肝微粒体蛋白含量为1132mg/g±143mg/g，CYP450含量为071nmol/mg±008nmol/mg，NADPH细胞色素C还原酶活性为301nmol/(min·mg)±026nmol/(min·mg)，AND活性为236nmol/(min·mg)±018nmol/(min·mg)，ERND活性为121nmol/(min·mg)±011nmol/(min·mg)，AH活性为025nmol/(min·mg)±003nmol/(min·mg)。结果表明，建立的鸡肝微粒体CYP450酶系的检测方法灵敏可靠，重复性较好。 |
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关键词：
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细胞色素P450； 肝微粒体； 鸡； 方法学 |
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中图分类号：S 859.7 文献标识码：
A 文章编号：1673-4696(2006)02-0147-04 |
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Methodological study on determination of activities of hepatic
microsomalcytochrome P450s in chickens |
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YANG Haifeng, JIANG Shanxiang |
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(College of Veterinary Medicine,Nanjing Agricultural University, Nanjing 210095, China) |
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Abstract: Twelve 60dayold cocks were selected to establish a method for determination of activities of hepatic microsomal cytochrome P450s. Liver microsome was distilled by different velocity centrifugation. The Bradford method was used to measure the concentration of protein in liver microsome. Activities of cytochrome P450s were measured by spectrophotometry. Protein concentration in hepatic microsome was 1132 mg/g±143 mg/g, content of cytochrome P450 was 071 nmol/mg±008 nmol/mg, activity of NADPHcytochrome C reductase was 301 nmol/(min·mg)±026 nmol/(min·mg),the activity of AND was 236 nmol/(min·mg)±018 nmol/(min·mg),the activity of ERND was 121 nmol/(min·mg)±011 nmol/(min·mg), and the activity of AH was 025 nmol/(min·mg)±003 nmol/(min·mg). Results suggested that the established method for determining contents and activities of hepatic microsomal cytochrome P450s in chickens was sensitive, reliable with good reproducibility. |
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Key words: cytochrome P450; liver microsome; chickens; methodology |
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