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为从药物酶的角度建立一种客观评价鱼类“首过效应”的方法,利用荧光定量PCR法测定尼罗罗非鱼(Oreochomis niloticus Linn)肝、肾组织中P-糖蛋白(P-gp)基因表达量,分析了单剂量(40 mg.kg 1)口服给药恩诺沙星(enrofloxacin,ENR)后,尼罗罗非鱼肠道、肝组织中mRNA水平的相对表达量与ENR血药浓度的时实相关性。实验结果显示:在尼罗罗非鱼肠道、肝组织中,P-gp基因在分子量127 bp处出现了与预期大小相符的特异性扩增片段。对尼罗罗非鱼口灌给药ENR后,ENR能迅速通过肠道进入血浆,其在肠道、肝和血浆中的消除速度较快,其药物时量曲线关系符合一级吸收的二室开放动力学模型。当血浆中ENR浓度达到最高达峰时(1 h),实验组肠道和肝中P-gp基因的相对表达量相对于对照组均表现出显著性差异(P<0.05);当肠道中ENR浓度达到最高峰时(2 h),实验组肠道P-gp基因的相对表达量则表现出极显著差异(P<0.01);当肝中ENR浓度达到最高峰时(2 h),实验组肝P-gp基因的相对表达量与对照组相比则表现出显著性差异(P<0.05)。该结果证实了鱼类P-gp基因参与药物代谢过程,提供了一种从分子水平揭示水产动物体内药物代谢规律的思路。
In order to establish a method of objective evaluation of fish “first pass effect” from the perspective of pharmaceutical enzymes, the P and glycoproteins (P and P) in liver and kidney tissues of Oreochromis niloticus Linn were determined by fluorescence quantitative PCR -gp) gene expression, the relative expression levels of mRNA in intestinal and hepatic tissues of Nile tilapia were analyzed after oral administration of enrofloxacin (ENR) with a single dose (40 mg.kg 1) ENR plasma concentrations of real-time correlation. The results showed that the P-gp gene appeared in the intestine and liver tissue of the Nile tilapia at a molecular weight of 127 bp at a specific amplified fragment corresponding to the expected size. ENR after oral administration of Nile tilapia, ENR can quickly enter the plasma through the intestine, its elimination in the intestine, liver and plasma faster, the relationship between the drug volume curve consistent with the absorption of the second room Open dynamics model. When the concentration of ENR peaked in plasma (1 h), the relative expression of P-gp gene in the intestine and liver of the experimental group showed significant difference compared with the control group (P <0.05). When the ENR When the concentration peaked (2 h), the relative expression of P-gp gene in experimental group showed a significant difference (P <0.01). When the ENR concentration reached its peak (2 h), the experimental group The relative expression of P-gp gene in liver showed a significant difference compared with the control group (P <0.05). This result confirmed the involvement of fish P-gp gene in the process of drug metabolism and provided a way to reveal the regularity of drug metabolism in aquatic animals from the molecular level.