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MiR‐214 Promotes the Alcohol‐Induced Oxidative Stress via Down‐Regulation of Glutathione Reductase and Cytochrome P450 Oxidoreductase in Liver Cells
Author(s) -
Dong Xiaolong,
Liu Hong,
Chen Fangjie,
Li Dejun,
Zhao Yanyan
Publication year - 2014
Publication title -
alcoholism: clinical and experimental research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.267
H-Index - 153
eISSN - 1530-0277
pISSN - 0145-6008
DOI - 10.1111/acer.12209
Subject(s) - oxidative stress , glutathione , malondialdehyde , cyp2e1 , hepatocyte , microbiology and biotechnology , chemistry , biochemistry , pharmacology , biology , cytochrome p450 , in vitro , metabolism , enzyme
Background The involvement of oxidative stress in the pathophysiological process of alcohol‐induced liver injury has been studied for decades. However, the role of micro RNA s (mi RNA s) targeting to oxidative stress genes in the pathogenesis of alcohol‐induced liver injury has not yet been determined. The aim of this study was to identify the targeting of miR‐214 to both glutathione reductase ( GSR ) and cytochrome P450 oxidoreductase ( POR ) genes and elucidate their impact on alcohol‐induced oxidative stress in liver cells. Methods The miR‐214 expression vector and reporter vectors of GSR and POR 3′‐ UTR were constructed. Human hepatoma cell (Bel7402), human embryonic kidney 293 cell ( HEK 293), and rat normal hepatocyte ( BRL ) were transfected and stimulated with ethanol (EtOH). Wistar rats were fed with EtOH for 4 weeks. The GSR and POR protein levels were detected by Western blot, and their activities were measured using the spectrophotometric method. The miR‐214 expression was detected by real‐time PCR . The index of oxidative stress including the total antioxidant capacity (T‐ AOC ) and malondialdehyde ( MDA ) level was detected by commercial kits. Results miR‐214 bound specifically to the GSR and POR 3′‐ UTR and repressed the expressions and activities of both GSR and POR . EtOH up‐regulated the miR‐214 expression, down‐regulated the GSR and POR protein levels and activities, and induced the oxidative stress in human and rat liver cells. EtOH‐fed Wistar rats further confirmed that alcohol up‐regulates the miR‐214 expression in liver and repressed both GSR and POR in vivo. Conclusions These findings demonstrated a new mechanism by which the alcohol repressed the GSR and POR expression via up‐regulation of miR‐214 and in turn induced oxidative stress in liver cells.