Protective Effect Of FGF21 On NAFLD Mice

Non-alcoholic fatty liver(NAFLD)is a high incidence disease at present in humans and animals,but it has not found a good way to treat it in clinic.Therefore,early prevention is an effective way to protect the liver from developing into non-alcoholic fatty liver(NAFLD).Fibroblast growth factor 21(FGF21)is a new metabolic regulator involved in lipid utilization.In order to clarify the protective effect of FGF21 on nonalcoholic fatty liver,as well as the changes of energy,inflammation and lipid were studied the protective effect of FGF21 in vivo and in vitro.Test one Study on exogenous inject FGF21 and its effect on non-alcoholic Fatty liver in HFD mice in vivo,the effect of FGF21 on immune response indicators and high cholesterol hematological parameters was investigated.In the experiment,a total of 24 female mice were selected,12 of which were fed with HFD and the other were fed with normal diet.After 30 days,the mice of two groups were respectively divided into four groups on average: 1.HFD mice were the HFD group;2.mice were the control group;3.mice were the HFD + FGF21 group;4.mice were the control+FGF21 group.The mice were killed on the third and sixth day after the injection,and the livers were isolated and analyzed.Blood was collected and analyzed for CD3 and CD19 by flow cytometry and IL-4,IL-6 and TNF-? were detected by ELISA.Serum TG,TC,NEFA,LDL-c,and HDL-c levels were measured by automatic biochemical analyzer.As a result,although HFD injection FGF21 group CD3 and CD19 significant reduce(P <0.01),and the levels of IL-4,IL-6,TNF-?,TC,TG LDL-c,and NEFA in HFD exogenous injection FGF21+HFD group were all lower than those in the HFD group(P <0.05).Test two To investigate the effect of FGF21 deficiency on non-alcoholic fatty liver in vitro,we used mice AML12 cells for testing,and used si RNA silencing FGF21,adenovirus to over express LKB1 for 12 hours,then added silencing and over-expression reagent to continue culture for 48 hours and 12 hours after starvation.Then 1.2mol/L NEFA was added to culture for12 hours,and the cells were collected to be examined.Lipid metabolism was evaluated by Laser confocal fluorescence of lipid droplets.Measurement of reactive oxygen species(ROS)by flow Cytometry.ELISA was used to detect FGF21,SIRT1,IL-4,IL-6,TNF-?.Biochemical analysis was used to detect GLU,TG,PA.WB was used to detect AMPK,p-AMPK.p-PCR wasused to detect FGF21,LKB1,SREBP-1c,IL-8,TNF-?.The result that silencing FGF21 obviously reduced the levels of AMPK phosphorylation through LKB1 in hepatocytes cells(P<0.05).Silencing FGF21 reduced the rate of p-AMPK/AMPK by inducing LKB1 and the expression of SREBP-1c downstream of AMPK(P <0.05).Glucose levels were significantly higher in NEFA-treated LKB1-overexpressing cells than in NEFA-treated cells(P <0.01).The glucose level in NEFA-treated FGF21-silenced cells was higher than that of NEFA-treated cells(P >0.05),but had no significant effect.When both exist at the same time,the expression of GLU is higher than that of NEFA-treated LKB1-overexpressing cells.IL-6 and TNF-? levels were significantly higher in NEFA-treated FGF21-silenced cells than in NEFA cells(P <0.05).The expression of ROS increased when FGF21 was silenced.Silencing FGF21 leads to the increase of lipid droplets,LKB1-overexpression appears to have no effect on lipids droplets.In summary,Exogenous FGF21 has a protective effect on mild non-alcoholic fatty liver in mice,which is reflected in the decrease of energy,immune index and inflammation.It was also proved by experiments in vitro that the increase of free fatty acids in the absence of FGF21,resulted in an increase in oxidative stress,abnormal energy,lipid metabolism and inflammation in hepatocytes.It is proved that FGF21 has a good prospect of protecting nonalcoholic fatty liver and provides theoretical and experimental basis for preventing the development of liver from developing into nonalcoholic fatty liver with high energy intake of FGF21 in the future.