Liver hepcidin mRNA expression is inappropriately low in alcoholic patients compared with healthy controls.
OBJECTIVE
Hepcidin plays a crucial role in iron metabolism, preventing its absorption at the basolateral enterocyte membrane. Hepcidin regulation is complex and regulated at the transcriptional level. The relation between iron overload and alcoholic liver disease is well known, but its mechanism is not clear. We present an observational, case-control study, aimed at evaluating the effects of alcohol on the expression of hepcidin in human participants. We intended to assess whether iron overload related to alcohol ingestion was caused by hepcidin-impaired expression by determining hepcidin mRNA expression and relating it to iron stores, both in alcoholic patients and in normal controls.


METHODS
We compared liver hepcidin mRNA expression between 25 active drinkers with alcoholic liver disease, without cirrhosis, and 20 healthy controls. All individuals were evaluated for HFE mutations, complete blood count, coagulation, glucose, kidney function, liver function, viral hepatitis, C-reactive protein, interleukin 6, tumor necrosis factor α, and serum iron, ferritin, and transferrin saturation. Total RNA was isolated from liver samples, cDNA was obtained by reverse transcription, and hepatic expression levels of hepcidin were determined by real-time PCR using the comparative Ct method (2(-ΔΔCt)).


RESULTS
Serum ferritin and transferrin saturation were significantly higher in patients. Hepcidin was downregulated in patients compared with the controls by a mean factor of -0.44 (log10 2(-ΔΔCt)) (P=0.009). Hepcidin expression was not significantly different between the several grades of fibrosis, necroinflammatory activity, and liver iron stores. Heavy alcohol consumption caused the highest hepcidin mRNA suppression. The hepcidin mRNA expression/serum ferritin ratio was significantly lower in alcoholic patients (P<0.0001).


CONCLUSIONS
Hepcidin liver expression is inappropriately low in alcoholic patients with active alcoholism and preserved hepatic function, and we conclude that this is the mechanism for alcohol consumption-associated iron overload in humans.
Faculty of Medicine, University of Coimbra, Coimbra, Portugal. costamatos.luis@gmail.com
Luís
L
Costa-Matos
Paulo
P
Batista
Nuno
N
Monteiro
Maria
M
Simões
Conceição
C
Egas
Jorge
J
Pereira
Helena
H
Pinho
Natália
N
Santos
João
J
Ribeiro
Maria A
MA
Cipriano
Pedro
P
Henriques
Fernando
F
Girão
Alfredo
A
Rodrigues
Armando
A
Carvalho
D000328:Adult; D000428:Alcohol Drinking / Q000009:adverse effects; D000437:Alcoholism / Q000150:complications* / Q000378:metabolism / Q000473:pathology; D000704:Analysis of Variance; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000378:metabolism*; D016022:Case-Control Studies; D000431:Ethanol / Q000009:adverse effects / Q000378:metabolism; D005260:Female; D005293:Ferritins / Q000032:analysis*; D064451:Hepcidins; D006801:Humans; D019190:Iron Overload / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D008099:Liver / Q000378:metabolism* / Q000503:physiopathology; D008108:Liver Diseases, Alcoholic / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D018709:Statistics, Nonparametric; D014168:Transferrin / Q000032:analysis*
D023181:Antimicrobial Cationic Peptides; C578702:HAMP protein, human; D064451:Hepcidins; D012333:RNA, Messenger; D014168:Transferrin; D000431:Ethanol; D005293:Ferritins