摘要
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11.32 mg/g±1.43 mg/g,CYP450含量为0.71 nmol/mg±0.08 nmol/mg,NADPH细-胞色素C还原酶活性为3.01 nmol/(min.mg)±0.26 nmol/(min.mg),AND活性为2.36 nmol/(min.mg)±0.18 nmol/(min.mg),ERND活性为1.21 nmol/(min.mg)±0.11 nmol/(min.mg),AH活性为0.25 nmol/(min.mg)±0.03nmol/(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。
Twelve 60-day-old 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 11.32 mg/g±1. 43 mg/g, content of cytochrome P450 was 0.71 nmol/mg±0.08 nmol/mg, activity of NADPH-cytochrome C reductase was 3.01 nmol/（min · mg）±0.26 nmol/（min · mg）,the activity of AND was 2.36 nmol/（min · mg）±0.18 nmol/（min · mg）,the activity of ERND was 1.21 nmol/（min · mg）± 0. 11 nmol/（min · mg）, and the activity of AH was 0.25 nmol/（min · mg）±0.03 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.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2006年第2期147-150,共4页
Chinese Veterinary Science
