Therapeuic Effects And Mechanisms Of P7C3-A20 In Non-alcoholic Fatty Liver Disease

Nonalcoholic fatty liver disease(NAFLD)is a Severe clinical syndrome caused by excessive fat deposition in liver but without the history of excessive drinkingor other specific hepatic impairment factors,whichhas related to insulin resistance and genetic susceptibility hepatic metabolic disease.This syndrome includes simple fatty liver(SFL),nonalcoholic steatohepatitis(NASH)and hepatic cirrhosis.In the United States,the prevalence of NAFLD among adults is about 30%.Domestic epidemiological studies have shown that NAFLD has become the second leading cause of liver dysfunction and chronic liver disease in China.Nicotinamide adenine dinucleotide(NAD),also known as Coenzyme?,could conduct hydrogen in the electron transport chain Complex?.The synthesis of NAD consists of two pathways,De novo pathway and Salvage pathway.The second one is the main way to maintain the level of NAD in cell.Nicotinamide phosphoribosyltransferases transferase(NAMPT)is the limited enzymein De novo pathway which maintainsthe NAD level.The synthesis of ATP request the activity of NAD,which is also widely involved in body growth,angiogenesis,apoptosis,and tissue inflammation.Previous studies on NAMPT function in our research group had found therelationship between NAMPT and NAFLD.The prevalenceof NAFLD in theaged mice maybe increased by the lower expression of NAMPT which is gradually decreased with increasing age.In addition,the decrease of NAMPT expression was important for the onset and progression of NAFLD in elderly mice.However,it is not clear whether NAMPT's related activators can treat NAFLD and the therapeutic mechanism.Therefore,the therapeutic effect and mechanism of compound P7C3-A20 on NAFLD requset furtherresearch.Methods: 1.The experimental animals were divided into four groups,and each group was not less than 6,including Chow group,Chow+A20 group,HFD group and HFD+A20 group.Chow group was fed with regular feed and HFD group was fed with high fat feed.The mice feeded with HFD in 16 W were induces by P7C3-A20 ip,with a dose of 20mg/kg/d.After giving the drug 2W,the treatment effect of P7C3-A20 on NAFLD was preliminarily evaluated from weight and metabolic rate among 4 groups.Then the serum lipid and liver enzyme system were detected.Finally,the effect of P7C3-A20 on NAFLD was detected from inflammatory factor,liver fibrosis process and apoptosis.2.There wereobvious changes of FGF1 and FGF21 in serum between HFD and HFD+A20 in our early stage.it is suggest that there are some potential relationship between P7C3-A20 and AMPK pathway activation.There are some specificmarker in AMPK pathway such as Akt,CREB and AMPK expresses in liver need to be tested.Mesntime the phosphorylation markers knowed asphosphorus Akt,phosphorus and phosphorus-CREB also need to be detected.These experimentsmay explore the specific sites of P7C3-A20 on AMPK pathway and explain the therapeuticmechanism.3.AMPK?2+/-mice were hybridized to prepare AMPK?2-/-mice.The WT mice were set as the control group and AMPK?2-/-mice were set as the experimental group.Both groups were given intraperitoneal injection of P7C3-A20,3 days,20mg/kg/d.The changes of FGF1 and FGF21 in serum were detected the expression of AMPK pathway related markers in the liver.This experiment may explore whether the treatment mechanism of P7C3-A20 for NAFLD is achieved through the AMPK pathway.Results: 1.HFD group was able to induce the NAFLD model.Secondly,after the HFD group gave P7C3-A20,compared with the control group of HFD,the mice showed significant reduction in weight,and the metabolic rate was increased.The indexes of liver enzyme in serum were improved,and FGF1 and FGF21 were significantly increased.Lipid deposition,cell inflammatory level and apoptosis of mouse liver were all decreased.Liver fibrosis and fiber composition were all improved,indicating that P7C3-A20 did have a therapeutic effect on NAFLD..2.The experiment with AMPK?2-/-mice suggest that P7C3-A20 could inhibited the up-regulation of markers on AMPK pathway,and both CREB and AMPK protein and phosphorylated protein showed no significant up-regulation.Compound P7C3-A20 of AMPK pathway after CREB and AMPK protein expression is raised,the phosphorylated proteins,namely phosphorus-CREB and phosphorus-AMPK is also raised.But Akt did not make a difference.Conclusion: 1.Compound P7C3-A20 has therapeutic effect on NAFLD.2.the AMPK pathway in the NAFLD animal model could be activated by P7C3-A20 and the expression of CREB and AMPK and phosphorylated protein were up-regulated.3.By AMPK?2-/-mouse validation.Compound P7C3-A20 was not able to up-regulate the expression of marker proteins and serum FGF1 and FGF21 in mouse.