Modulation of miR-26a-5p and miR-15b-5p exosomal expression associated with clopidogrel-induced hepatotoxicity in HepG2 cells

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dc.contributor.authorde Freitas, Renata C. Costa
dc.contributor.authorBortolin, Raul H.
dc.contributor.authorLopes, Mariana B.
dc.contributor.authorTamborlin, Letícia
dc.contributor.authorMeneguello, Letícia [UNESP]
dc.contributor.authorSilbiger, Vivian N.
dc.contributor.authorHirata, Rosario D.C.
dc.contributor.authorHirata, Mário H.
dc.contributor.authorLuchessi, Augusto D. [UNESP]
dc.contributor.authorLuchessi, André D.
dc.contributor.institutionFederal University of Rio Grande do Norte
dc.contributor.institutionUniversidade Estadual de Campinas (UNICAMP)
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.date.accessioned2018-12-11T17:23:43Z
dc.date.available2018-12-11T17:23:43Z
dc.date.issued2017-12-12
dc.description.abstractClopidogrel is an essential antiplatelet drug used to prevent thrombosis complications associated with atherosclerosis. However, hepatotoxicity is a potential adverse effect related to clopidogrel therapy. Exosome-derived miRNAs may be useful for improved monitoring of drug response and hepatotoxicity risk. In the present study, the expression of several exosomal miRNAs (miR-26a-5p, miR-145-5p, miR-15b-5p, and miR-4701-3p) and cell-derived mRNA targets (PLOD2, SENP5, EIF4G2, HMGA2, STRADB, and TLK1) were evaluated in HepG2 cells treated with clopidogrel (6.25, 12.5, 25, 50, and 100 μM) for 24 and 48 h. Then, clopidogrel cytotoxicity was evaluated by analyzing DNA fragmentation and the cell cycle profile using flow cytometry. Differential expression of exosome-derived miRNAs and cell-derived mRNAs was analyzed by RT-qPCR. Exposure of HepG2 cells to high concentrations of clopidogrel (50 and 100 μM) for 24 h caused significant DNA fragmentation (17.6 and 44.4%, respectively; p < 0.05) and 48 h (26.8 and 48.9%, respectively; p < 0.05), indicating cellular toxicity. Upregulation of miR-26a-5p and downregulation of miR-15b-5p was observed in cells exposed to 100 μM clopidogrel for 24 and 48 h. The miR-26a-5p target mRNAs HMGA2, EIF4G2, STRADB, and SENP5 were downregulated in HepG2 cells following exposure to cytotoxic concentrations of clopidogrel (50 and 100 μM) for 24 h, and HMGA2 levels remained low after 48 h of treatment. TLK1, a target of miR-15b-5p, was downregulated by 50 and 100 μM clopidogrel at 24 h. In conclusion, our results suggest that exposure to high concentrations of clopidogrel modulates the expression of exosomal miR-26a-5p and miR-15b-5p and their target mRNAs in HepG2 cells. Dysregulation of these miRNAs maybe modulate the regulatory pathways involved in clopidogrel-induced liver injury.en
dc.description.affiliationDepartment of Clinical and Toxicological Analysis Federal University of Rio Grande do Norte
dc.description.affiliationLaboratory of Biotechnology School of Applied Sciences University of Campinas
dc.description.affiliationPost graduation in Biological Science Institute of Biosciences São Paulo State University (UNESP)
dc.description.affiliationDepartment of Clinical and Toxicological Analyses School of Pharmaceutical Sciences University of São Paulo
dc.description.affiliationUnespPost graduation in Biological Science Institute of Biosciences São Paulo State University (UNESP)
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipIdFAPESP: 2010/18095-0
dc.description.sponsorshipIdCNPq: 447120/2014-0
dc.description.sponsorshipIdCNPq: 448753/2014-6
dc.identifierhttp://dx.doi.org/10.3389/fphar.2017.00906
dc.identifier.citationFrontiers in Pharmacology, v. 8, n. DEC, 2017.
dc.identifier.doi10.3389/fphar.2017.00906
dc.identifier.file2-s2.0-85037836568.pdf
dc.identifier.issn1663-9812
dc.identifier.scopus2-s2.0-85037836568
dc.identifier.urihttp://hdl.handle.net/11449/177068
dc.language.isoeng
dc.relation.ispartofFrontiers in Pharmacology
dc.relation.ispartofsjr1,587
dc.rights.accessRightsAcesso aberto
dc.sourceScopus
dc.subjectClopidogrel
dc.subjectHepatotoxicity
dc.subjectHepG2 cell line
dc.subjectMicroRNAs
dc.titleModulation of miR-26a-5p and miR-15b-5p exosomal expression associated with clopidogrel-induced hepatotoxicity in HepG2 cellsen
dc.typeArtigo

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