Abstract: ObjectiveTo establish a rat model of alcoholic liver injury and to explore the effect of D-limonene on iron overload in rats with chronic alcoholic liver injury.MethodsSeventy male wistar rats were fed adaptively for seven days and then randomly divided into seven groups(n=10 for each group),including a control group,a alcohol-model group(alcoholic liver damage),the low,medium and high D-limonene treated groups(100,200,400 mg·kg^-1·day^-1),a limonene control group(400 mg/kg),and a glycyrrhizic acid group.The experiment lasted for six weeks.The histopathology of the liver was observed after hematoxylin-eosin(HE)staining.Ultrastructural changes of the livers were observed with transmission electron microscope(TEM).The liver iron content was measured with inductively coupled plasma-mass spectrometry(ICP-MS).In addition,the levels of plasma hepcidin,plasma ferritin and plasma hemojuvelin(HJV)were assessed with enzyme-linked immunosobent assay(ELISA).ResultsIn the alcohol-model group,the structure disturbance of hepatic lobule,the diffuse fatty degeneration in the liver tissue,and obvious hyperplasia of the hepatic fibrous tissue with visible spotty necrosis were observed.Compared with those of the alcohol-model group,the degree of structure disturbance in hepatic lobule,liver fatty degeneration and damage were all significantly reduced in various doses D-limonene groups.TEM showed that hepatic ultrastructure lesions,including the structure of endoplasmic reticulum,mitochondria and lipid droplet,were reduced in different doses D-limonene groups compared with alcohol-model group.The rats in alcohol-model group had a significant increase in liver iron content(279.14±5.87 μg/mL)and level of plasma ferritin(57.76±5.83 ng/mL)(P<0.05) compared with those of the normal group;but both the level of hepcidin(61.85±4.85 ng/mL)and HJV(46.21±5.24 ng/mL)were obviously decreased(P<0.05).Compared with those of the model group,both the level of liver iron(180.14±5.76 μg/mL)and plasma ferritin(35.39±3.94 ng/mL)were decreased(P<0.05) and the level of hepcidin(81.36±4.8 ng/mL)and HJV(67.29±5.89 ng/mL) were increased in high dose D-limonene treatments groups(P<0.05).ConclusionD-limonene can restrain iron overload in the rats exposed to ethanol.