摘要
目的研究睾酮在人及小鼠、大鼠、犬、兔、猪、豚鼠、牛、羊等实验动物肝微粒体代谢中的差异。方法睾酮分别于上述种属肝微粒体进行孵育,加入NADP Na、葡萄糖-6-磷酸、葡萄糖-6-磷酸脱氢酶等试剂启动体外P450酶反应。样品采用高效液相色谱紫外检测器及高分辨质谱进行分析。代谢轮廓用热图进行可视化,并用主成分分析进行差异研究。结果 6β-羟基化不是睾酮唯一的代谢途径。本研究在不同种属中共发现了18种睾酮的代谢产物。尽管6β-羟基化在不同种属中与睾酮的底物消除基本呈正相关,但该途径无法补偿其他途径造成的底物消除。豚鼠具有最强的睾酮代谢能力,而羊对睾酮的代谢能力最弱。小鼠、犬、兔肝微粒体中,睾酮的代谢轮廓与人最相似。结论 6β羟基化途径的定量不能用作所有物种CYP 3A活性评判的精准依据。如果药物证明由CYP 3A4进行代谢,在临床前期研究中,应结合药理学模型的需要,尽量采用小鼠、犬、兔作为动物模型,以保证药物体内暴露与人体实验尽量接近。
Objective To investigate the species difference of testosterone metabolism among human and 7 animal species including mice, rats, guinea pigs, rabbits, dogs, pigs, sheep, and cattle. Methods Testosterone was incubated with liver microsomes of humans and investigated animals, together with CYP - mediated metabolic reaction factors including NADP Na, glucose - 6 - phosphate, and glucose - 6 - phosphate dehydrogenase. Incubation samples were introduced into HPLC to determinate testosterone and its metabolites. Heatmap was used to visualize the metabolic profile while principle component analysis was used to tell the difference of metabolic profile among species. High resolution mass spectrometry was used to assist the identification of metabolite structures. Results 6 ~ - hydroxylation is not the unique metabolic pathway of testosterone. Eighteen NADP - dependent metabolites were observed in humans or other investigated animals. Although 6~ -hydroxylation showed positive correlation with the substrate elimination, this specific metabolic pathway could not compensate all substrate consumption. Guinea pig generates the most metabolites while sheep showed the weakest capacity on testosterone metabolism. Mice, dogs, and rabbits showed the most similar metabolic profile to humans. Conclusion 6 β - hydroxylation metabolites of testosterone was not applicable to evaluate the CYP 3A activity of all species. Mice, dogs, and rabbits are recommended in pre - clinical trial if the target is proved to be metabolized by human CYP 3A4.
出处
《遵义医学院学报》
2017年第5期463-468,共6页
Journal of Zunyi Medical University
基金
国家自然科学基金资助项目(NO:81402985
NO:81560673
NO:81660685)
贵州省科技重大专项(NO:黔科合重大专项字NO:[2015]6010)
贵州省科学技术基金资助项目(NO:黔科合J字[2015]2158
黔科合JZ字[2015]2010)
贵州省出国留学人员择优资助计划项目(NO:[2015]03)
国家级/省级大学生创新创业项目(NO:201510661002)
贵州省药剂学研究生卓越人才培养计划(黔教研合ZYRC字[NO:2014]019)
遵义医学院博士启动基金
遵义市科技局联合基金(遵市科合社字NO:[2016]15)
关键词
睾酮
种属差异
CYP
3A
代谢
testosterone
species difference
CYP 3A
metabolism
