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Impact on acetaminophen‐induced hepatotoxicity by long non‐coding RNAs HNF1α‐AS1 and HNF4α‐AS1 in HepaRG cells
Author(s) -
Chen Liming,
Manautou Jose E.,
Zhong Xiaobo
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.506.6
Subject(s) - gene knockdown , chemistry , acetaminophen , pharmacology , cyp2e1 , glutathione , cytochrome p450 , biochemistry , biology , apoptosis , enzyme
Acetaminophen (APAP) is a commonly used over‐the‐counter drug for its analgesic and antipyretic effects. However, APAP overdose leads to severe hepatotoxicity due to the accumulation of N ‐acetyl‐ p ‐benzoquinone imine (NAPQI), the toxic metabolite of APAP generated by cytochrome P450s (P450s). Long non‐coding RNAs HNF1α‐antisense 1 (HNF1α‐AS1) and HNF4α‐antisense 1 (HNF4α‐AS1) are regulatory RNAs, which are recently identified to be involved in the regulation of hepatic P450 expression. This study aims to determine whether HNF1α‐AS1 and HNF4α‐AS1 have impacts on APAP‐induced hepatotoxicity using the most useful hepatic cell line HepaRG as an in vitro model. Small hairpin RNAs (shRNAs) were used to knock down HNF1α‐AS1 and HNF4α‐AS1 in HepaRG cells. Knocking down of these lncRNAs altered APAP‐induced hepatotoxicity indicated by MTT and LDH cytotoxicity assays. Specifically, HNF1α‐AS1 knockdown decreased APAP toxicity at 10 mM with increased cell viability and decreased LDH release, whereas HNF4α‐AS1 knockdown exacerbated APAP toxicity at 30 mM. Alterations of gene expression by knocking down HNF1α‐AS1 and HNF4α‐AS1 were examined in several APAP‐induced hepatotoxicity pathways, including P450‐mediated APAP biotransformation by CYP1A2, 2E1, and 3A4, APAP detoxification with glucuronidation by UDP‐glucuronosyltransferases, UGT1A1 and 1A9, and sulfation by sulfotransferases SULT1A1, and NAPQI detoxification through glutathione conjugation by glutathione‐ S ‐transferases, GSTP1 and GSTT1. HNF1α‐AS1 knockdown significantly decreased mRNA levels of CYP1A2, 2E1, and 3A4, while HNF4α‐AS1 knockdown significantly induced mRNA levels of CYP2E1 and 3A4 in HepaRG cells. HNF1α‐AS1 knockdown decreased mRNA expression of SULT1A1 but not UGT1A1 or 1A9, while HNF4α‐AS1 knockdown showed no significant effect on the selected phase II enzymes. Significantly decreased mRNA expression of GSTP1 was observed after either HNF1α‐AS1 or HNF4α‐AS1 knockdown. However, considering the low intrinsic expression of GSTP1 in HepaRG cells, how this decreased expression of GSTP1 affects APAP toxicity is inconclusive. GSTT1 expression showed no change in all groups. Altogether, our study suggests that HNF1α‐AS1 and HNF4α‐AS1 affect APAP‐induced hepatotoxicity mainly through alterations of P450‐mediated APAP biotransformation process in HepaRG cells, indicating an important role of lncRNAs in drug‐induced liver toxicity. Support or Funding Information This work was supported by grant NIH/NIGMS R01GM118367. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .