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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300μl(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)×103nmol/(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。
Objective To study the effect of multi-house echinococcosis infection on hepatic drug metabolism enzyme activity in Meriones unguiculatus. Methods Ten Meriones unguiculatus mice were randomly divided into two groups. Each experimental group was inoculated intraperitoneally with 300μl of echinococcosis cyst tissue (about 600 protoscoleces), and the control group was given equal volume of normal saline . Five months after infection, gerbils were sacrificed by cervical dislocation and the liver was removed. The suspension and cytosol of liver microsomes were prepared by differential centrifugation. The bicinchoninic acid (BCA) protein quantitation kit was used to determine the protein concentration of the cytosolic and microsomal suspensions. The contents of cytochrome P450 (CYP450) and cytochrome b5 (Cyt b5) in liver microsomes were determined by differential spectroscopy. Determination of the activity of 7-ethoxyresorufin deacetylase (EROD) and 7-methoxyhalodrine demethylase (MROD) by fluorescence spectrophotometry. The activity of NADPH-cytochrome C reductase (NCR), glutathione S-transferase (GST) and flavone monooxygenase (FMO) were determined by UV-Vis spectrophotometry. Results The protein concentration of the suspension and liver microsome in the experimental group were (11.089 ± 1.277) and (3.212 ± 0.924) mg / ml, respectively, and that in the control group were (12.459 ± 1.625) and (3.894 ± 0.395) mg / ml, respectively. The contents of CYP450 and Cyt b5 in experimental group were (0.508 ± 0.142) and (0.515 ± 0.077) nmol / mg, which were significantly lower than those in control group [(0.647 ± 0.090) and (0.596 ± 0.051) nmol / (P <0.05). The activity of GST in experimental group was (1.766 ± 0.339) × 103 nmol / (mg · min), which was significantly lower than that in control group [(2.001 ± 0.160) × 103 nmol / (mg · min)] The activities of FMO and NCR in the liver microsome were (1.142 ± 0.327) and (0.602 ± 0.162) × 103 nmol / (mg · min), respectively, which were significantly higher than those in the control group [(0.882 ± 0.150) and (0.442 ± 0.082) × (P <0.05). However, the activity of EROD and MROD in liver microsomes was not significantly different from that in control group (P> 0.05). Conclusion The results showed that the activity of FMO and NCR in liver microsomes increased obviously and the activity of GST decreased significantly in gerbils infected with Echinococcus multilocularis.