MiR-155 and other microRNAs downregulate drug metabolizing cytochromes P450 in inflammation
MiR-155 and other microRNAs downregulate drug metabolizing cytochromes P450 in inflammation
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DOI：
10.1016/j.bcp.2019.113725
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发表时间：
2020-01-01
影响因子：
5.8
通讯作者：
Zanger, Ulrich M.
中科院分区：
文献类型：
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作者：
Kugler, Nicole;Klein, Kathrin;Zanger, Ulrich M.
在急性和慢性炎症条件下，由于药物代谢酶和转运蛋白的协同下调，肝脏的解毒能力严重受损。通过对C反应蛋白(CRP)升高的供者肝组织进行整体转录组学分析，我们观察到阳性和阴性急性时相反应的程度相当，其中最高上调的基因集包括免疫反应和防御途径，而下调主要发生在代谢和分解代谢途径，包括许多重要的药物代谢酶和转运蛋白。我们假设microRNAs(MiRNA)，通常作为基因表达的负调控者，参与了这一过程。微阵列和实时定量聚合酶链式反应分析发现，来自CRP升高、胆汁淤积症、脂肪变性或非酒精性脂肪性肝炎的供者的肝组织中存在差异表达的miRNAs。利用含有天然和突变的3‘-未翻译基因区的荧光素酶报告基因构建，验证了RXRα(miR-130b-3p)、CYP2C8(miR-452-5p)、CYP2C9(miR-155-5p)、CYP2C19(miR-155-5p、miR-6807-5p)和CYP3A4(miR-224-5p)上的几个预测的miRNA结合位点。转染miRNA模拟物的HepaRG细胞显示出mRNA水平和几种细胞色素P450酶活性的协同降低，特别是miR-155-5p、miR-452-5p和miR-6807-5p，这是在所有四种病理条件下唯一解除调控的miRNA。此外，我们观察到肝组织miRNA水平与肝脏CYP表型之间存在强烈的负相关关系。由于miR-155以其在免疫、炎症和癌症中的多功能作用而闻名，我们的数据表明，该miR-155和其他miRNAs有助于在炎症条件下协调下调药物代谢酶和转运蛋白。
In conditions of acute and chronic inflammation hepatic detoxification capacity is severely impaired due to coordinated downregulation of drug metabolizing enzymes and transporters. Using global transcriptome analysis of liver tissue from donors with pathologically elevated C-reactive protein (CRP), we observed comparable extent of positive and negative acute phase response, where the top upregulated gene sets included immune response and defense pathways while downregulation occurred mostly in metabolic and catabolic pathways including many important drug metabolizing enzymes and transporters. We hypothesized that microRNAs (miRNA), which usually act as negative regulators of gene expression, contribute to this process. Microarray and quantitative real-time PCR analyses identified differentially expressed miRNAs in liver tissues from donors with elevated CRP, cholestasis, steatosis, or non-alcoholic steatohepatitis. Using luciferase reporter constructs harboring native and mutated 3'-untranslated gene regions, several predicted miRNA binding sites on RXR alpha (miR-130b-3p), CYP2C8 (miR-452-5p), CYP2C9 (miR-155-5p), CYP2C19 (miR-155-5p, miR-6807-5p), and CYP3A4 (miR-224-5p) were validated. HepaRG cells transfected with miRNA mimics showed coordinate reductions in mRNA levels and several cytochrome P450 enzyme activities particularly for miR-155-5p, miR-452-5p, and miR-6807-5p, the only miRNA that was deregulated in all four pathological conditions. Furthermore we observed strong negative correlations between liver tissue miRNA levels and hepatic CYP phenotypes. Since miR-155 is well known for its multifunctional roles in immunity, inflammation, and cancer, our data suggest that this and other miRNAs contribute to coordinated downregulation of drug metabolizing enzymes and transporters in inflammatory conditions.