miR-150对药物代谢酶CYP3A4的调控作用

doi:10.12092/j.issn.1009-2501.2018.07.005

中国临床药理学与治疗学 ›› 2018, Vol. 23 ›› Issue (7): 749-754.doi: 10.12092/j.issn.1009-2501.2018.07.005

miR-150对药物代谢酶CYP3A4的调控作用

刘莉¹，彭金富²，郭成贤²，阳喜定³，李海刚¹，阳国平²

¹长沙医学院药学院，长沙 410200，湖南；²中南大学湘雅三医院，长沙 410013；³中南大学湘雅二医院，长沙 410011，湖南

收稿日期:2017-12-20 修回日期:2018-04-10 出版日期:2018-07-26 发布日期:2018-07-20
通讯作者: 阳国平，男，教授，硕士生导师，主要从事临床药理学与药代动力学研究。 Tel：0731-88618339 E-mail：ygp9880@163.com
作者简介:刘莉，女，助教，主要从事临床药理学与药代动力学研究。 Tel：13469079764 E-mail：liuli2013good@163.com
基金资助:
湖南省教育厅科学研究项目资助（17C0169）；国家自然科学基金（81373476）；新型药物制剂研发湖南省重点实验室培育基地资助（2016TP1029）；湖南省教育厅科学研究项目资助（15C0160）

Regulative effects of miR-150 on CYP3A4

LIU Li¹, PENG Jinfu², GUO Chengxian², YANG Xiding³, LI Haigang¹, YANG Guoping²

¹Department of Pharmacy, Changsha Medical University, Changsha 410200, Hunan, China; ²Department of Pharmacy, the Third Xiangya Hospital, Central South University, Changsha 410013, Hunan, China; ³ Department of Pharmacy, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan, China

Received:2017-12-20 Revised:2018-04-10 Online:2018-07-26 Published:2018-07-20

摘要/Abstract

摘要：

目的: 观察miR-150对药物代谢酶CYP3A4的调控作用。方法: 用不同浓度游离长链脂肪酸（FFA）刺激张氏肝细胞，使之成为脂肪变性肝细胞。通过荧光定量PCR和Western blot测定CYP3A4 mRNA和蛋白表达以及miR-150表达。生物信息学分析并构建野生型CYP3A4 3′-UTR双荧光素酶报告载体，与miR-150模拟物共转染。另外，在张氏肝细胞和脂肪变性肝细胞中分别转染miR-150模拟物和抑制剂，检测CYP3A4 mRNA和蛋白表达。结果: 张氏肝细胞经1 mmol/L FFA诱导24 h后，与对照组相比，脂肪变性肝细胞中CYP3A4 mRNA和蛋白表达均显著下降（P<0.05），miR-150表达上升（P<0.05）。经生物信息学分析发现CYP3A4为miR-150的靶基因，转染质粒和miR-150模拟物后，荧光活性下降。miR-150模拟物可使CYP3A4 mRNA表达呈浓度梯度下降（P<0.05），也可使CYP3A4蛋白表达下降；miR-150抑制剂使CYP3A4 mRNA表达上升（P<0.05）。结论: miR-150可直接与CYP3A4的3'-UTR结合，进而直接调控CYP3A4的表达。

关键词: miR-150, CYP3A4, 调控作用

Abstract:

AIM: To explore the role of miR-150 in the regulation of CYP3A4 and the underlying mechanism. METHODS: CYP3A4 mRNA and protein expression were tested by RT-PCR and Western blot when the Chang liver cells were stimulated by FFA containing fatty acids free 1% BSA. Bioinformatics miRNA databases were used to identify CYP3A4-related miRNAs and the wild-type CYP3A4 3′-UTR plasmids were constructed, miR-150 and reporter plasmid were co-transfected into the Chang liver cells to test the expression ratio of the reported fluorescence and the correction fluorescence. Then the miR-150 mimic were transfected into the Chang liver cells, while miRNA inhibitor were transfected into the steatosis cells. RT-PCR and Western blot were used to detect CYP3A4 mRNA and protein expression. RESULTS: The CYP3A4 mRNA and protein level were statistically significantly decreased（P<0.05）, while the mature miR-150 levels were increased (P<0.05）when the Chang liver cells were stimulated after 24 h by 1 mmol/L FFA. CYP3A4 was found to be the target gene of miR-150 with a bioinformatics analysis. Then CYP3A4 mRNA level was significantly decreased (P<0.05). Meanwhile, the expression level of CYP3A4 protein also decreased when the miR-150 mimic were transfected into the Chang liver cells. But CYP3A4 mRNA expression has no statistical significance (P=0.071) in miR-150 inhibitor group. CONCLUSION: The miR-150 can combine with CYP3A4 3′-UTR directly, and can further regulate the expression of CYP3A4 mRNA.

Key words: miR-150, CYP3A4, regulation

中图分类号:

R965.2

刘莉，彭金富，郭成贤，阳喜定，李海刚，阳国平. miR-150对药物代谢酶CYP3A4的调控作用[J]. 中国临床药理学与治疗学, 2018, 23(7): 749-754.

LIU Li, PENG Jinfu, GUO Chengxian, YANG Xiding, LI Haigang, YANG Guoping. Regulative effects of miR-150 on CYP3A4[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(7): 749-754.

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miR-150对药物代谢酶CYP3A4的调控作用

Regulative effects of miR-150 on CYP3A4

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