M1 polarization bias and subsequent nonalcoholic steatohepatitis progression is attenuated by nitric oxide donor DETA NONOate via inhibition of CYP2E1-induced oxidative stress in obese mice

J Pharmacol Exp Ther. 2015 Jan;352(1):77-89. doi: 10.1124/jpet.114.218131. Epub 2014 Oct 27.


Activation of M1 macrophages in nonalcoholic steatohepatitis (NASH) is produced by several external or endogenous factors: inflammatory stimuli, oxidative stress, and cytokines are known. However, any direct role of oxidative stress in causing M1 polarization in NASH has been unclear. We hypothesized that CYP2E1-mediated oxidative stress causes M1 polarization in experimental NASH, and that nitric oxide (NO) donor administration inhibits CYP2E1-mediated inflammation with concomitant attenuation of M1 polarization. Because CYP2E1 takes center stage in these studies, we used a toxin model of NASH that uses a ligand and a substrate of CYP2E1 for inducing NASH. Subsequently, we used a methionine and choline-deficient diet-induced rodent NASH model where the role of CYP2E1 in disease progression has been shown. Our results show that CYP2E1 causes M1 polarization bias, which includes a significant increase in interleukin-1β (IL-1β) and IL-12 in both models of NASH, whereas CYP2E1-null mice or diallyl sulfide administration prevented it. Administration of gadolinium chloride (GdCl3), a macrophage toxin, attenuated both the initial M1 response and the subsequent M2 response, showing that the observed increase in cytokine levels is primarily from macrophages. Based on the evidence of an adaptive NO increase, the NO donor administration in vivo that mechanistically inhibited CYP2E1 catalyzed the oxidative stress during the entire study in NASH-abrogated M1 polarization and NASH progression. The results obtained show the association of CYP2E1 in M1 polarization, and that inhibition of CYP2E1 catalyzed oxidative stress by an NO donor (DETA NONOate [(Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate]) can be a promising therapeutic strategy in NASH.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cytochrome P-450 CYP2E1 / genetics
  • Cytochrome P-450 CYP2E1 / metabolism*
  • Disease Progression*
  • Gene Expression Regulation / drug effects
  • Lipid Peroxidation / drug effects
  • Liver / drug effects
  • Liver / metabolism
  • Macrophages / cytology
  • Macrophages / drug effects*
  • Macrophages / immunology
  • Male
  • Mice
  • Mice, Obese
  • Nitric Oxide Donors / pharmacology
  • Nitric Oxide Donors / therapeutic use
  • Nitroso Compounds / pharmacology*
  • Nitroso Compounds / therapeutic use
  • Non-alcoholic Fatty Liver Disease / drug therapy*
  • Non-alcoholic Fatty Liver Disease / immunology
  • Non-alcoholic Fatty Liver Disease / metabolism*
  • Non-alcoholic Fatty Liver Disease / pathology
  • Oxidative Stress / drug effects*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Tyrosine / metabolism


  • Nitric Oxide Donors
  • Nitroso Compounds
  • RNA, Messenger
  • 2,2'-(hydroxynitrosohydrazono)bis-ethanamine
  • Tyrosine
  • Cytochrome P-450 CYP2E1