Granzyme B and miR-378a Interaction in Acetaminophen Toxicity in Children.
Granzyme B and miR-378a Interaction in Acetaminophen Toxicity in Children.
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DOI：
10.2174/2211536608666190808144456
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发表时间：
2020
影响因子：
--
通讯作者：
Gill, Pritmohinder
中科院分区：
文献类型：
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作者：
McCullough, Sandra;Dweep, Harsh;McGill, Mitchell R;Bhattacharyya, Sudeepa;James, Laura;Frankowski, Sara;Woodall, Aaron;Kearns, Gregory;Gill, Pritmohinder
肝脏I期药物代谢酶CYP2E1， CYP1A2和CYP3A4催化对乙酰氨基酚（APAP）的生物转化，在毒性介导中起重要作用。其他肝脏和非肝脏I期酶在APAP毒性中的潜在作用尚未确定。将APAP过量住院患儿分为无毒性（ALT≤45 IU/L, n=5）和中度毒性（ALT≥500 IU/L, n=5）亚组，对包含84个I期药物代谢相关基因的PCR阵列进行检测。无毒性亚组ALDH6A1、CYP4F12和GZMB显著下调，中毒性亚组ALDH1A1、CYP27A1和GZMB显著下调。qRT-PCR证实ALDH1A1、CYP4F12和GZMB显著下调。芯片分析发现GZMB 3'UTR是miR-378a-5p的靶标。据报道，过表达miR-378a-5p可使GZMB 3'UTR报告质粒的荧光素酶活性降低50%。与模拟对照相比，转染miR-378a-5p模拟物的NK-92细胞延长了APAP对GZMB蛋白表达的影响。此外，在接受NAC治疗的APAP过量儿童血液样本中，miR-378a-5p显著上调。总的来说，我们的研究表明存在一种新的信号通路，miR-378a-5p抑制APAP过量儿童GZMB的表达。
Hepatic phase I drug-metabolizing enzymes CYP2E1, CYP1A2 and CYP3A4 catalyze the biotransformation of Acetaminophen (APAP) and are important in the mediation of toxicity. The potential role of other hepatic and non-hepatic Phase I enzymes in APAP toxicity has not been established. PCR array containing 84 genes involved in phase I drug metabolism was examined in subgroups of hospitalized children for APAP overdose, categorized as no toxicity (ALT ≤ 45 IU/L, n=5) and moderate toxicity (ALT ≥ 500 IU/L, n=5). Significant downregulation was observed for ALDH6A1, CYP4F12 and GZMB in the no toxicity subgroup and ALDH1A1, CYP27A1 and GZMB in the moderate toxicity subgroup. qRT-PCR confirmed significant downregulation for ALDH1A1, CYP4F12, and GZMB. In-silico analysis identified GZMB 3’UTR to be a target of miR-378a-5p. Overexpression of miR-378a-5p reduced the luciferase activity of GZMB 3’UTR reporter plasmid reportedly by 50%. NK-92 cells transfected with the miR-378a-5p mimic extended the effect of APAP on GZMB protein expression compared to mimic controls. In addition, miR-378a-5p was significantly upregulated in blood samples of children with APAP overdose undergoing NAC treatment. Overall, our study suggests the presence of a novel signaling pathway, whereby miR-378a-5p inhibits GZMB expression in children with APAP overdose.