| ObjectivesMethionine deficiency(MD)and choline deficiency(CD)can cause non-alcoholic fatty liver disease(NAFLD),but the mechanism is obviously different.The animal model of Non-alcoholic steatohepatitis(NASH)induced by combined methionine and choline deficiency(MCD)is one of the commonly used model methods to study the prevention and treatment of NAFLD.But the role of methionine deficiency and choline deficiency in this model is still unclear.This article compared MD,CD,MCD-induced NAFLD models in fatty liver,liver oxidative stress,lipid peroxidation and liver The change of elements,so as to explore the role of MD and CD in the NASH model induced by MCD,provides an important theoretical basis for the mechanism research of NAFLD.MethodsForty male adult C57 BL / 6J mice(25 ± 5g)were randomly divided into 4 groups and fed with MD diet,CD diet,MCD diet and Methionine and choline sufficient(MCS)control diet.After 1 week of adaptive feeding,the model group and the control group were fed with corresponding diets.After 8 weeks,fasting(12h),weighing and blood samples were collected under anesthesia,followed by saline perfusion and execute,the whole liver was collected,the appearance of the liver was observed,and the liver wet weight was measured.After preparing serum from blood samples,use automatic biochemical analyzer to determine alanine aminotransferase(ALT),aspartate transaminase(AST),serum iron(SI),serum triglycerides(serum-TG),serum total cholesterol(serum-TC).Liver tissues were fixed with 4% paraformaldehyde fixative solution,used for hematoxylin-eosin(HE)staining and oil red O fat staining,observe the sample tissue morphology and fatty degeneration.some liver tissues were homogenized treatment,use the kit to determine the total antioxidant capacity(T-AOC),malondialdehyde(MDA),triglyceride(TG),Total cholesterol(T-CHO)and free fatty acid(FFA),using inductively coupled plasma emission spectrometer(ICP-OES)to measure non-heme iron(NHI)of liver tissue and liver iron,liver copper,liver zinc,liver manganese,liver selenium,liver molybdenum element content.ResultsGeneral situation: MD and CD had no interactive effects on the body weight of NASH mice induced by MCD(P > 0.05),and the main effect of MD(P < 0.01),which is similar to the MCS group compared with that,the weight of mice in MD and MCD groups decreased significantly(P < 0.01),and there was no statistically significant change in the weight of mice in CD group(P > 0.05).Compared with MD group,there was no significant change in the weight of mice in MCD group significance(P > 0.05).compared with the CD group,the body weight of the MCD group was significantly reduced(P < 0.01).The effects of MD and CD on MCD-induced liver wet weight and liver index of NASH mice have an interactive effect(P < 0.01),both of which are antagonistic.The main effect of MD has a main effect(P < 0.001),which is the same as that of MCS group.Compared with the MD,CD and MCD group,the liver wet weight and liver index were significantly reduced(P < 0.01).Compared with that of the CD group,the liver wet weight of the MCD group was significantly reduced(P < 0.01).Compared with that of the MD group,the liver index of the MCD group was obviously increased(P < 0.05).Liver pathological slices: The results of liver histopathological slices showed that the mice in the MCD group had ballooning fat vacuoles in the liver,while the mice in the MD and CD groups had no ballooning fat vacuoles.Serum indicators: MD and CD have an interactive effect on the serum ALT,AST,SI,serum-TC,serum-TG of NASH mice induced by MCD(P < 0.01),both of which show a synergistic effect,and the effects of MD and CD are both there is a main effect(P < 0.05).Compared with that of the MCS group,the changes of serum ALT,AST,and SI in the MD and CD groups were not statistically significant(P > 0.05),and the ALT,AST,and SI in the MCD group were significantly increased(P < 0.01).Compared with that of the MD group,the ALT,AST,and SI of the MCD group were significantly increased(P < 0.01).Compared with that of the CD group,the ALT,AST,and SI of the MCD group were significantly increased(P < 0.01).Compared with that of the MCS group,Serum-TG in the MCD group was significantly reduced(P < 0.01),and Serum-TC in the MD,CD,and MCD groups was significantly reduced(P < 0.01).Compared with that of the MD and CD groups,the MCD group was smaller rat serum-TC was significantly decreased(P < 0.01).Liver oxidative stress indicators: MD and CD have an interactive effect on mouse liver T-CHO and MDA(P < 0.05),showing a synergistic effect,and there is a main effect on the effects of MD and CD(P < 0.05).Compared with that of the MCS group,the changes of T-AOC and MDA in the MD and CD groups were not statistically significant(P > 0.05),the liver T-AOC of the MCD group was significantly reduced(P < 0.01),and the MDA was significantly increased(P < 0.001).Liver steatosis indicators: MD and CD have an interactive effect on the liver TG,T-CHO and FFA of mice(P < 0.01),both of which show a synergistic effect,and the effects of MD and CD both have a main effect(P < 0.05).Compared with that of the MCS group,the liver of TG,T-CHO,and FFA in the MD group were significantly increased(P < 0.01),and the change of liver TG,T-CHO,and FFA in the CD group were not statistically significant(P > 0.05),liver TG,T-CHO and FFA of MCD group were significantly increased(P < 0.001).Compared with that of MD group,liver TG of MCD group was significantly increased(P < 0.01).Compared with that of CD group compared with that in MCD group,liver TG,T-CHO and FFA contents were significantly increased(P < 0.001).Liver trace elements: MD and CD have interaction effects on liver NHI,liver copper,liver zinc,liver manganese,and liver molybdenum in mice(P < 0.01).Synergistic effect,antagonistic effect on liver zinc,MD has a main effect on NHI,liver copper,liver manganese,liver molybdenum(P < 0.05),CD has a main effect on liver copper,liver manganese(P < 0.05).MD and CD have no interaction effect on liver iron in MCD-induced NASH mice(P > 0.05),and MD has a main effect on liver iron(P < 0.001).Compared with that of the MCS group,the liver NHI and liver iron of the MD and MCD groups were significantly increased(P < 0.01),and the changes of liver NHI and liver iron of the CD group were not statistically significant(P >0.05).and Compared with that of the MD group,the liver NHI and liver iron of the MCD group were not statistically significant(P > 0.05).Compared with that of the CD group,the liver NHI and liver iron of the MCD group were significantly increased(P < 0.01).Compared with that of the MCS group,the changes of liver manganese,liver copper,liver zinc and liver molybdenum concentrations in the MD and CD groups were not statistically significant(P > 0.05),and the liver manganese in the MCD group was significantly increased(P < 0.01),liver copper,liver zinc and liver molybdenum were significantly reduced(P < 0.05).ConclusionThe role of MD is to reduce the weight,liver wet weight,liver index,serum-TC,serum-TG,and liver T-AOC,liver copper concentration,liver molybdenum concentration,and increase serum ALT,serum AST,SI,and liver MDA,TG,T-CHO,FFA,NHI,liver iron,liver copper content of mice induced by MCD.The role of CD is to reduce serum-TG,serum-TC,liver T-AOC and liver copper concentration,and increase serum ALT,serum AST,SI,liver MDA,TG,T-CHO,FFA,and Liver manganese concentration of mice induced by MCD.MD and CD on MCD-induced NASH mouse serum ALT,AST,SI,Serum-TG,Serum-TC,liver TG,T-CHO,FFA,liver T-AOC,MDA,liver NHI,liver copper,liver manganese,The effect of hepatic molybdenum is synergistic.the performance of MCD-induced NASH mice liver wet weight,liver index,liver zinc is antagonistic.the effect on body weight and liver iron concentration is MD alone. |
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