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中国临床药理学与治疗学 ›› 2004, Vol. 9 ›› Issue (5): 500-503.

• 研究原著 • 上一篇    下一篇

大鼠肝微粒体细胞色素P450 酶系检测方法学研究

朱曼, 王睿, 张永青, 梁蓓蓓   

  1. 解放军总医院临床药理药学研究室, 北京100853
  • 收稿日期:2003-11-29 修回日期:2004-02-11 发布日期:2020-11-22
  • 通讯作者: 朱曼, 女, 硕士研究生, 药师。Tel:010-66937141  E-mai l:zhangwenyuan @chinasatcom.com

Study on hepatic cytochromes P450 methodology in Wistar rats

ZHU Man, WANG Rui, ZHANG Yong-Qing, CHEN Kun   

  1. Department of Clinical Pharmacology, Chinese PLA General Hospital, Beijing 100853, China
  • Received:2003-11-29 Revised:2004-02-11 Published:2020-11-22

摘要: 目的 建立大鼠肝微粒体蛋白含量以及肝微粒体细胞色素P450 酶系含量与活性测定的紫外和荧光分光光度方法。 方法 应用差速离心法提取大鼠肝微粒体, Lowery 法测定肝微粒体蛋白浓度, 应用紫外和荧光分光光度法测定肝微粒体细胞色素P450 酶系含量及活性。 结果 牛血清白蛋白标准曲线的线性范围为25 ~ 250 mg·L-1, 最低检测限为25 mg·L-1, 相关系数r 为0.9975;紫外分光光度法测定细胞色素P450 和细胞色素b5 含量及NADPHCytC还原酶活性的结果显示方法灵敏;测定氨基比林N-脱甲基酶、红霉素N-脱甲基酶活性的甲醛标准曲线, 线性范围为0.05~0.5 mmol·L-1, 最低检测限为0.05 mmol·L-1, 相关系数r 为0.9988;测定7-乙氧基香豆素脱烃酶活性的resorufin 标准曲线线性范围为1~8 μmol·L-1, 最低检测限为1 μmol·L-1,相关系数r 为0.9998。 结论 紫外和荧光分光光度方法测定大鼠肝微粒体细胞色素P450 酶系中6 种酶的含量及活性的灵敏可靠, 重复性较好。

关键词: 紫外分光光度计, 荧光分光光度计, 肝微粒体细胞色素P450

Abstract: AIM: To study the methodology of hepatic cytochromes P450 in Wistar rats. METHODS: After the Wistar rats were administered with gatifloxacin and ciprofloxacin once daily for 7 days, and liver microsome was distilled by different velocity centrifugation.The Lowry method was used to measure the concentration of protein in liver microsome, and the activities of gatifloxacin in hepatic microsomal enzymes (CYP450)of rats were studied by spectrophotometer. RESULTS: The linearity range of ox blood serum albumin standard curve was 25-250 mg·L-1 with the detection limits 25 mg·L-1 and correlation coefficient 0.9988.The contents of cytochromes P450 and cytochromes b5 and the activity of NADPH-cytochrome C reductase indicated that ultraviolet spectrophotometer method was sensitive.The linearity range of formaldehyde standard curve, which was used to measure the activities of aminopyrine-N-demethylase and erytheomycin-N-demethylase was 0.05-0.5 mmol ·L-1 with the detection limits 0.05mmol·L-1 and correlation coefficient 0.9998.The linearity range of resorufin standard curve, which was used to measure the activity of Coumarin 7-hydroxylation was 1-8 μmol·L-1 with the detection limits 1 μmol·L-1 and correlation coefficient 0.9998. CONCLUSION: The methods of measuring the contents and activities of the six hepatic microsomal enzymes are sensitive and credible in Wistar rats by ultraviolet and fluorescence spectrophotometer.

Key words: ultraviolet spectrophotometer, fluorescence spectrophotometer, cytochromes P450

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