中国寄生虫学与寄生虫病杂志

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多房棘球蚴感染对长爪沙鼠肝脏药物代谢酶活性的影响

李焕平,辛奇,鲁俊,袁苗苗,Nabeel PERVAIZ,景涛*   

  1. 兰州大学基础医学院病原生物学研究所,兰州 730000
  • 出版日期:2016-02-28 发布日期:2016-03-11

Effect of Echinococcus multilocularis Infection on the Activities of Drug-metabolizing Enzymes in Gerbil Liver

LI Huan-ping, XIN Qi, LU Jun, YUAN Miao-miao, Nabeel PERVAIZ, JING Tao*   

  1. Research Institute of Pathogenic Biology, School of Basic Medicine, Lanzhou University, Lazhou 730000, China
  • Online:2016-02-28 Published:2016-03-11

摘要:

目的 研究多房棘球蚴感染对长爪沙鼠肝脏药物代谢酶活性的影响。 方法 将10只长爪沙鼠随机均分为2组,实验组每鼠腹腔接种多房棘球蚴囊组织匀浆300 μl(约含600个原头节),对照组每鼠给予等量生理盐水。感染后5个月,颈椎脱臼法处死沙鼠,取出肝脏,以差速离心法制备肝微粒体悬液和细胞液。采用二喹啉甲酸(BCA)蛋白定量分析试剂盒测定细胞液和微粒体悬液的蛋白浓度。用差示光谱法测定肝微粒体中细胞色素P450(CYP450)和细胞色素b5(Cyt b5)的含量。荧光分光光度法测定7-乙氧基试卤灵脱乙基酶(EROD)和7-甲氧基试卤灵脱甲基酶(MROD)的活性。紫外-可见光分光光度法测定NADPH-细胞色素C还原酶(NCR)、谷胱甘肽巯基转移酶(GST)和黄素单加氧酶(FMO)的活性。 结果 实验组细胞液和肝微粒体悬液的蛋白浓度为(11.089±1.277)和(3.212±0.924)mg/ml,对照组的分别为(12.459±1.625)和(3.894±0.395)mg/ml。实验组肝微粒体CYP450和Cyt b5的含量为(0.508±0.142)和(0.515±0.077)nmol/mg蛋白,明显低于对照组[(0.647±0.090)和(0.596±0.051)nmol/mg蛋白](P<0.05)。实验组细胞液GST活性为(1.766±0.339)×103 nmol/(mg·min),明显低于对照组[(2.001±0.160)×103 nmol/(mg·min)](P<0.05);实验组肝微粒体FMO和NCR的活性分别为(1.142±0.327)和(0.602±0.162)×103 nmol/(mg·min),明显高于对照组[(0.882±0.150)和(0.442±0.082)×103 nmol/(mg·min)](P<0.05);而肝微粒体EROD和MROD的活性与对照组相比,差异无统计学意义(P>0.05)。 结论 长爪沙鼠感染多房棘球蚴后,肝微粒体FMO和NCR活性明显升高,GST活性明显降低。

关键词: 多房棘球蚴, 长爪沙鼠, 药物代谢酶, 比色法

Abstract:

Objective To study the effect of Echinococcus multilocularis infection on the activities of drug-metabolizing enzymes in gerbil liver. Methods Ten gerbils (Meriones unguiculatus) were randomly divided into 2 groups, the experimental group in which each animal was intraperitoneally injected with 300 μl E. multilocularis cyst homogenates (containing about 600 protoscoleces), and the control group in which each animal received 300 μl saline(i.p.). Five months after infection, the animals were sacrificed by cervical dislocation, and livers were collected. The liver microsomal and cytosolic fractions were obtained by differential centrifugation. Protein concentration was measured by the BCA method. CYP450 and Cyt b5 contents in the microsomal fraction were measured with differential spectroscopy. Activities of 7-ethoxyresorufin(EROD) and 7-methoxyresorufin(MROD) were measured by fluorescence spectrometry. The activities of NADPH-cytochrome C reductase(NCR), glutathione-S-transferase (GST), and flavine monooxygenases (FMO) were measured by UV-visible spectrophotometry. Results The protein content of cytosolic fractions and liver microsomes in experimental group was (11.089±1.277) and (3.212±0.924) mg/ml, those in control group was (12.459±1.625) and (3.894±0.395) mg/ml, respectively. The contents of CYP450 and Cyt b5 in the experimental group [(0.508±0.142), (0.515±0.077) nmol/mg protein, respectively] were both significantly lower than those in the control [(0.647±0.090), (0.596±0.051) nmol/mg protein](P<0.05). The GST activity decreased significantly in the experimental group [(1.766±0.339)×103 nmol/(mg·min)] compared with the control [(2.001±0.160)×103 nmol/(mg·min)](P<0.05). But the FMO and NCR activities increased significantly in the experimental group [(1.142±0.327) nmol/(mg·min) and (0.602±0.162)×103 nmol/(mg·min), respectively] compared with the control [(0.882±0.150) nmol/(mg·min) and (0.442±0.082)×103 nmol/(mg·min)](P<0.05). However, there were no significant differences in EROD and MROD activities between the two groups(P>0.05). Conclusion The activities of FMO and NCR markedly increase, while that of GST significantly decreases in the gerbil liver after E. multilocularis infection.

Key words: Echinococcus multilocularis, Meriones unguiculatus, Drug-metabolizing enzyme, Colorimetry