• cytochromes P-450
• delta- [or 5-] aminolevulinic acid [synthase]
• heme
• liver cell cultures
• liver cell organoids
• porphyria
• porphyrins

• heme iron absorption
• iron transport
• iron uptake in duodenum
• non-heme iron absorption

• Baicalin
• Curcumin
• Nrf2/HO‐1 pathway
• ethanol
• hepatic toxicity
• oxidative stress

• 2017-7-12 Science and Technology Bureau, Chengguan District, Lanzhou, Gansu, China
• 2017-RC-79 Talent Innovation and Entrepreneurship Project, Lanzhou, Gansu, China

• 5-aminolevulinic acid
• Binge Alcohol Exposure Liver Injury
• Heme Oxygenase-1
• Inflammation
• Macrophages

This study was designed to explore the role of cytochrome P4502E1 (CYP2E1) expression in the course of brain edema induced by subacute poisoning with 1,2-dichloroethane (1,2-DCE). Mice were randomly divided into five groups: the control group, the 1,2-DCE poisoned group, and the low-, medium- and high-dose diallyl sulfide (DAS) intervention groups. The present study found that CYP2E1 expression levels in the brains of the 1,2-DCE-poisoned group were upregulated transcriptionally; in contrast, the levels were suppressed by DAS pretreatment in the intervention groups. In addition, the expression levels of both Nrf2 and HO-1 were also upregulated transcriptionally in the brains of the 1,2-DCE-poisoned group, while they were suppressed dose-dependently in the intervention groups. Moreover, compared with the control group, MDA levels and water contents in the brains of the 1,2-DCE-poisoned group increased, whereas NPSH levels and tight junction (TJ) protein levels decreased significantly. Conversely, compared with the 1,2-DCE- poisoned group, MDA levels and water contents in the brains of the intervention groups decreased, and NPSH levels and TJ protein levels increased significantly. Furthermore, pathological changes of brain edema observed in the 1,2-DCE-poisoned group were markedly improved in the intervention groups. Collectively, our results suggested that CYP2E1 expression could be transcriptionally upregulated in 1,2-DCE-poisoned mice, which might enhance 1,2-DCE metabolism in vivo, and induce oxidative damage and TJ disruption in the brain, ultimately leading to brain edema.

• 1
• 2-DCE)
• 2-dichloroethane (1
• brain edema
• cytochrome P450 2E1 (CYP2E1)
• diallyl sulfide (DAS)
• heme oxygenase-1 (HO-1)
• nuclear factor erythroid 2-related factor 2 (Nrf2)

Excess alcohol consumption is a global crisis contributing to over 3 million alcohol-related deaths per year worldwide and economic costs exceeding $200 billion dollars, which include productivity losses, healthcare, and other effects (e.g., property damages). Both clinical and experimental models have shown that excessive alcohol consumption results in multiple organ injury. Although alcohol metabolism occurs primarily in the liver, alcohol exposure can lead to pathophysiological conditions in multiple organs and tissues, including the brain, lungs, adipose, liver, and intestines. Understanding the mechanisms by which alcohol-mediated organ dysfunction occurs could help to identify new therapeutic approaches to mitigate the detrimental effects of alcohol misuse. Hypoxia-inducible factor (HIF)-1 is a transcription factor comprised of HIF-1α and HIF-1β subunits that play a critical role in alcohol-mediated organ dysfunction. This review provides a comprehensive analysis of recent studies examining the relationship between HIF-1α and alcohol consumption as it relates to multiple organ injury and potential therapies to mitigate alcohol’s effects.

• HIF-1α
• alcohol
• ethanol

• 5-Aminolevulinic acid
• Brain
• Heme metabolism
• Porphobilinogen deaminase
• Volatile anaesthetics, porphyrinogenic drugs

AIM: To understand the role of toll-like receptor 4 (TLR4) signaling in the regulation of iron-regulatory hormone, hepcidin by chronic alcohol consumption.

METHODS: For chronic alcohol intake studies, TLR4 mutant mice on C3H/HeJ background and wildtype counterpart on C3H/HeOuJ background were pair-fed with regular (control) and ethanol-containing Lieber De Carli liquids diets. Gene expression was determined by real-time quantitative PCR. Protein-protein interactions and protein expression were determined by co-immunoprecipitation and western blotting. The occupancy of hepcidin gene promoter was determined by chromatin immunoprecipitation assays.

RESULTS: Chronic alcohol intake suppressed hepcidin mRNA expression in the livers of wildtype, but not TLR4 mutant, mice. The phosphorylation and nuclear translocation of nuclear factor (NF)-κB p65 subunit protein was observed in alcohol-fed wildtype, but not in alcohol-fed TLR4 mutant, mice. Similarly, alcohol induced the binding of NF-κB p50 subunit protein to hepcidin gene promoter in wildtype, but not in TLR4 mutant, mice. In contrast, the phosphorylation of Stat3 in the liver was stronger in alcohol-treated TLR4 mutant mice compared to alcohol-treated wildtype mice. The occupancy of hepcidin gene promoter by Stat3 was observed in alcohol-fed mutant, but not in wildtype, mice. An interaction between NF-κB p65 subunit protein and small heterodimer partner protein (SHP) was observed in the livers of both wildtype and TLR4 mutant mice fed with the control diet, as shown by co-immunoprecipitation studies. Alcohol intake elevated cytosolic SHP expression but attenuated its interaction with NF-κB in the liver, which was more prominent in the livers of wildtype compared to TLR4 mutant mice.

CONCLUSION: Activation of TLR4 signaling and NF-кB are involved in the suppression of hepcidin gene transcription by alcohol in the presence of inflammation in the liver.

• Iron
• Small heterodimer partner protein
• Nuclear factor-κB
• Alcoholic liver disease
• Inflammation

• Animals
• Binding Sites
• Disease Models, Animal
• Down-Regulation
• Ethanol
• Hepcidins/genetics
• Hepcidins/metabolism
• Liver/metabolism
• Liver Diseases, Alcoholic/etiology
• Liver Diseases, Alcoholic/genetics
• Liver Diseases, Alcoholic/metabolism
• Male
• Mice, Inbred C3H
• Mice, Mutant Strains
• Mutation
• Phosphorylation
• Promoter Regions, Genetic
• RNA, Messenger/metabolism
• STAT3 Transcription Factor/metabolism
• Signal Transduction
• Toll-Like Receptor 4/genetics
• Toll-Like Receptor 4/metabolism
• Transcription Factor RelA/metabolism
• Transcription, Genetic

AIM: To understand the role of mitochondrial-produced superoxide (O₂^•-) in the regulation of iron-regulatory hormone, hepcidin by alcohol in the liver.

METHODS: For alcohol experiments, manganese superoxide dismutase knockout mice heterozygous for Sod2 gene expression (Sod2^+/-) and age-matched littermate control mice (LMC), expressing Sod2 gene on both alleles, were exposed to either 10% (w/v) ethanol in the drinking water or plain water (control) for 7 d. Total cellular O₂^•- levels in hepatocytes isolated from the livers of mice were measured by electron paramagnetic resonance spectroscopy. The mitochondrial-targeted, O₂^•--sensitive fluorogenic probe, MitoSOX Red and flow cytometry were utilized to measure O₂^•- in mitochondria. Gene and protein expression were determined by Taqman Real-time quantitative PCR and Western blotting, respectively.

RESULTS: Sod2^+/- mice expressed 40% less MnSOD protein (SOD2) in hepatocytes compared to LMC mice. The deletion of Sod2 allele did not alter the basal expression level of hepcidin in the liver. 10% ethanol exposure for 1 wk inhibited hepatic hepcidin mRNA expression three-fold both in Sod2^+/- and LMC mice. O₂^•- levels in hepatocytes of untreated Sod2^+/- mice were three-fold higher than in untreated LMC mice, as observed by electron paramagnetic resonance spectroscopy. O₂^•- levels in mitochondria of Sod2^+/ mice were four-fold higher than in mitochondria of untreated LMC mice, as measured by MitoSOX Red fluorescence and flow cytometry. Alcohol induced a two-fold higher increase in O₂^•- levels in hepatocytes of LMC mice than in Sod2^+/- mice compared to respective untreated counterparts. In contrast, 1 wk alcohol exposure did not alter mitochondrial O₂^•- levels in both Sod2^+/- and control mice.

CONCLUSION: Mitochondrial O₂^•- is not involved in the inhibition of liver hepcidin transcription and thereby regulation of iron metabolism by alcohol. These findings also suggest that short-term alcohol consumption significantly elevates O₂^•- levels in hepatocytes, which appears not to originate from mitochondria.

• Hepcidin
• Alcohol
• Iron
• Superoxide
• Superoxide dismutase
• Liver
• Mitochondria
• Manganese superoxide dismutase
• Electron paramagnetic resonance

• P30 GM103335 NIGMS NIH HHS
• R01 AA017738 NIAAA NIH HHS