The Protective Effect Of Algin On Liver Of Hepatolenticular Degeneration In Mice

Objective:Hepatolenticular degeneration（HLD）is a kind of autosomal recessive inherited diseases induced by copper metabolic abnormality,and no special efficient therapies has been found presently in the world.In this study,Kunming mice were given a copper sulfate-loaded diet to establish the mouse models of HLD,and to o explore the protective effect of Algin on Liver of HLD mice.Methods:70 adult healthy male Kunming mice,aged 4-6 weeks,weighted 25-30g,were adaptively feed in the animal room,natural light,free to eat and drink water for 1 week.Then 10 animals were randomly selected as a control group,and the remaining 60animals were established with HLD models by copper sulfate-loaded diet method.Then,50 successful HLD models were included in the statistical range and randomly divided into control group,model group,positive drug group,low-dose treatment group,medium-dose treatment group,high-dose treatment group consisting of 10 models in each group respectively.The Algin was given by gavage for 4 consecutive weeks according to the following method:The mice of low-dose,medium-dose and high-dose treatment groups were orally given Algin as 50 mg/kg,100 mg/kg and 200 mg/kg body weight respectively each day for 4 weeks;the mice of positive drug group were orally given sodium dimercaptosuccinate（20 mg/kg body weight）as synchronization each day for 4weeks;while the mice of control and model groups were orally given same amount of normal solution at the same time for 4 weeks.After treatment,the changes of consciousness,reaction,movement,fur and hair,diet,feces and urine of mice were observed.The mouse urine was continuously collected with metabolic cage for 24 h.The concentration of copper ion in urine were determined by inductively coupled plasma-mass spectrometry（ICP-MS）.The hepatic functional indexes including activeities of alanine transaminase（ALT）and aspartate transaminase（AST）,concentrations of total protein（TP）and albumin（ALB）in serum detected by automatic biochemical analyzer.The pathological changes of hepatocytes were observed by HE staining and Masson staining was used to observe the hepatic fibrosis.The apoptosis of hepatocytes was detected by TUNEL assay and the immunohistochemical assay was used to determine the expression level of ceruloplasmin（CER）in hepatocytes.Results:The concentration of copper ion in urine increased significantly（P<0.01）.The activities of ALT and AST of hepatic functional indexes were elevated while the serum levels of TP and ALB reduced significantly than those in the control group（P<0.01）.The hepatocytes occurred degeneration and apoptosis,while the CER down-expressed significantly in hepatocytes（P<0.05）.After treatment with Algin,the spirit,behavioral functions of mice in high-dose group improved significantly than those in the model group.The concentration of copper ion in urine and the hepatic functional indexes ALT and AST activities in serum were decreased significantly than those in the model group（P<0.05）.The degenerated cell indexes（DCI）,hepatic fibrosis degrees（MOD）and apoptotic cell indexes（ACI）decreased,while the expression level of CER in hepatocytes（PCI）increased significantly than those in the model group（P<0.05）.Conclusions:Algin could play a protective effect for liver by enhancing CER expression and promoting Cu²⁺excretion to reduce the numbers of degenerative and apoptotic hepatocytes and the degrees of hepatic fibrosis,so that to improve the hepatic functions of HLD mince.The therapeutic effect of high-dose Algin（200mg/kg）was better than that of medium and low dose Algin.