Li Xiaoyan, Lu Yiyan, Ou Xiaojun, Zeng Sijing, Wang Ying, Qi Xiaoxiao, Zhu Lijun, Liu Zhongqiu
International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, China.
State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China.
PeerJ. 2020 Nov 18;8:e10182. doi: 10.7717/peerj.10182. eCollection 2020.
This study aimed to systematically profile the alterations and sex- and age-related differences in the drug metabolizing enzymes (DMEs) in a KRAS-mutant mouse model of lung cancer (KRAS mice).
In this study, the LC-MS/MS approach and a probe substrate method were used to detect the alterations in 21 isoforms of DMEs, as well as the enzymatic activities of five isoforms, respectively. Western blotting was applied to study the protein expression of four related receptors.
The proteins contents of CYP2C29 and CYP3A11, were significantly downregulated in the livers of male KRAS mice at 26 weeks (3.7- and 4.4-fold, respectively, < 0.05). SULT1A1 and SULT1D1 were upregulated by 1.8- to 7.0- fold at 20 ( = 0.015 and 0.017, respectively) and 26 weeks ( = 0.055 and 0.031, respectively). There were positive correlations between protein expression and enzyme activity for CYP2E1, UGT1A9, SULT1A1 and SULT1D1 ( ≥ 0.5, < 0.001). Western blotting analysis revealed the downregulation of AHR, FXR and PPARα protein expression in male KRAS mice at 26 weeks. For sex-related differences, CYP2E1 was male-predominant and UGT1A2 was female-predominant in the kidney. UGT1A1 and UGT1A5 expression was female-predominant, whereas UGT2B1 exhibited male-predominant expression in liver tissue. For the tissue distribution of DMEs, 21 subtypes of DMEs were all expressed in liver tissue. In the intestine, the expression levels of CYP2C29, CYP27A1, UGT1A2, 1A5, 1A6a, 1A9, 2B1, 2B5 and 2B36 were under the limitation of quantification. The subtypes of CYP7A1, 1B1, 2E1 and UGT1A1, 2A3, 2B34 were detected in kidney tissue.
This study, for the first time, unveils the variations and sex- and age-related differences in DMEs in C57 BL/6 (WT) mice and KRAS mice.
本研究旨在系统分析肺癌KRAS突变小鼠模型（KRAS小鼠）中药物代谢酶（DME）的变化以及与性别和年龄相关的差异。
在本研究中，采用液相色谱-串联质谱法（LC-MS/MS）和探针底物法分别检测21种DME同工型的变化以及5种同工型的酶活性。应用蛋白质印迹法研究4种相关受体的蛋白表达。
在26周龄的雄性KRAS小鼠肝脏中，CYP2C29和CYP3A11的蛋白含量显著下调（分别为3.7倍和4.4倍，P<0.05）。SULT1A1和SULT1D1在20周龄（分别为P = 0.015和0.017）和26周龄（分别为P = 0.055和0.031）时上调1.8至7.0倍。CYP2E1、UGT1A9、SULT1A1和SULT1D1的蛋白表达与酶活性之间存在正相关（r≥0.5，P<0.001）。蛋白质印迹分析显示，26周龄雄性KRAS小鼠中AHR、FXR和PPARα蛋白表达下调。关于性别差异，在肾脏中CYP2E1以雄性为主，UGT1A2以雌性为主。在肝脏组织中，UGT1A1和UGT1A5的表达以雌性为主，而UGT2B1则以雄性为主。关于DME的组织分布，21种DME亚型均在肝脏组织中表达。在肠道中，CYP2C29、CYP27A1、UGT1A2、1A5、1A6a、1A9、2B1、2B5和2B36的表达水平低于定量限。在肾脏组织中检测到CYP7A1、1B1、2E1以及UGT1A1、2A3、2B34亚型。
本研究首次揭示了C57 BL/6（野生型）小鼠和KRAS小鼠中DME的变化以及与性别和年龄相关的差异。
