Study Of Ethanol-Induced Chondriosome Injury And Alcoholic Liver Disease

Background: In west countries, alcohol abuse is a leading cause of serious liver disease. The incidence of ALD in our country has increased markedly and alcohol becomes the second reason of hepatic lesion after virus hepatitis in these years. The classical clinical manifestations of ALD are alcoholic fatty liver, alcoholic hepatitis and alcoholic liver fibrosis, which even can become inconvertible alcoholic cirrhosis. So the prevention and cure of ALD become more and more important in medical topic. The study and research of ALD has increasingly increased in world, but most of them are concentrating on the direct toxic action to liver cell by alcohol itself or its metabolic product and the contribution by the changes of nutritional disturbance, liver metabolism abnormality and lipid peroxidation in ALD. Immune reaction, cytogene abnormal regulation and apoptosis taking part in the process of pathological lesion in ALD were found in these years and apoptosis was considered as the major factor in the pathogenesis of ALD. The mechanism of liver cell apoptosis has not been well known, even though some research display that liver cell apoptosis is connected with activation of CYP2E1, addition of iron burden, augment of activity of TNF or TNF receptor, oxidative stress with oxide increasing and anti-oxide decreasing, apoptosis effect induced by myofibroblast and TGF β and other apoptosis factors. Some researches found that hepatocyte mitochondria swelled and its function deteriorated. The usage of chronic alcohol can inhibit the duplication of mtDNA, thus the number of mitochondra in hepatocyte is reduced. The content of GSH in mitochondra of ALD was found decrease obviously in some other studies, just because of the changes in the structure and function of the mitochondra play an important part in the function and damage of hepatocyte. So our investigation that the alcoholic disease should be concentrate on the mechanism and the pathway of the damage that to the mitochondrium.Objective: By setting up rat models of alcoholic fatty liver, to determin the oxidation and anti-oxidation index, and to observe the ultramicro-structure changes of mitochondrium in liver cells. Investigate the changes of PTP, ΔΨm, Ca²⁺ ,which are the correlated indexes when the alcohol induce the apoptosis, to approach the effect and mechanism of alcoholic disease through mitochondria pathway, thus to reveal a new view of mechanism of alcoholic liver disease and to provide experiment basis for clinical therapy.Materials and Methods: Forty male Wistar rats weighting between 140g and 160g were divided into two groups randomly: model group and control group,20 in model group were given alcoholic intragastric administration plus Oliver oil diet.20 in control group were given equal normal saline. By use the method of alcohol infusion to the stomach and olive oil balanced diet, we have succeeded establishing the models of ALD. The ultramicro-structure changes of mitochondria were observed under electron microscope; Extract mitochondria of liver, and detect patency of permeability transition pore (PTP) ; Isolate and putify hepatocyte, and marked them with fluorochrome rhodamine123 (Rh123) , Noyl-Acrydine Orange ( NAO ) and Fluo-3/AM,then Mitochondrial membrane potential (ΔΨm), Cardioplin in mitochondria membrane and [Ca²⁺]i in liver cell were measured by flow cytometry (FCM).Results: By use the method of alcohol infusion to the stomach and olive oil balanced diet , we have succeeded establishing the models of alcoholic disease. Membrane broken and cristae broken or disappeared were observed in mitochondria in different shapes under electron microscope. Some mitochondria show U shape or megamitochondrion. Compared with the control group, rats in the model group : (1) liver mitochondria PTP broke, and mitochondria swelled, A₅₄₀ decreased(p<0.01); (2) Decreased mitochondria membrane potential (p<0.01); (3) Decreased Cardioplin in mitochondria membrane(P<0.01); (4) Increased [Ca²⁺]i in liver cell(P<0.01);Conclusions: Alcohol intake might lead to mitochondria PTP broke, decreased mitochondria membrane potential, impair mitochondria membrane , and mitochondria Ca²⁺ out flow in liver cell ,.Ethanol-induced chondriosome injury may be an important mechanism of alcoholic disease.