Study On The Effect And Mechanism Of Tibetan Medicine “Ese” On Non-alcoholic Fatty Liver Disease With Typy Ⅱ Diabetes

Compared with the general people,patients with type Ⅱ diabetes are more likely to suffer from non-alcoholic fatty liver disease(NAFLD).Diet is an important part of the pathogenesis and treatment process of type Ⅱ diabetes combined with NAFLD.Therefore,the development of functional food is of great significance for the prevention and treatment of diabetes combined with non-alcoholic fatty liver disease.As a traditional noble drink in Tibet,the medicine Malus toringoides(Rehd.)Hughes(E se)is also an important medicine for the clinical treatment of hyperglycemia.In the preliminary study,Malus toringoides(Rehd.)Hughes has shown hypolipidemic and anti-oxidant effects,and has proved the plant contains abundant phlorizin,which has hypoglycemic activity.However,the treatment of type Ⅱ diabetes with NAFLD by Malus toringoides(Rehd.)Hughes has not been reported.Therefore,in this study,the pharmacological effects of Malus toringoides(Rehd.)Hughes extract(MT)on type Ⅱ diabetes mellitus combined with NAFLD were studied in vitro and in vivo,and the mechanism of its pharmacological effects was preliminarily explored to provide theoretical basis for its application in metabolic disorders.The model of diabetic rats with NAFLD was established by high glucose and high fat diet(HFD)and streptozotocin(STZ)injection.The body weight and blood glucose of rats were measured every week.The oral glucose tolerance(OGTT)and insulin tolerance(ITT)were measured at the 8~thh week.After that,rats were killed,serum and liver samples were collected,and biochemical indexes in serum and liver were detected by kit and ELISA.The pathological changes of liver were observed by H&E,Oil red O and PAS staining.NAFLD cell model was established by inducing HepG2cells with FFA.The indexes related to lipid accumulation and inflammation were detected by kit and ELISA,and the lipid content was observed by Oil red O staining.Western blot,immunohistochemistry and immunofluorescence were used to detect the effect of MT on the expression of SREBP-1c and NF–κB related proteins in vitro and in vivo.The data showed that the MT could reduce the hyperglycemia of diabetic rats,and significantly improve the OGTT,ITT,GSP,insulin and other indicators of diabetic rats.In addition,the liver lipid accumulation,oxidative stress,inflammation and other related indicators of the MT group were improved.The results of H&E,PAS and Oil red O staining also showed that MT significantly improved the liver lipid accumulation of diabetic rats Accumulation and inflammation.The results of NAFLD cell model established by HepG2 cells induced by FFA in vitro indicated that MT had significant effect on the indexes of lipid accumulation and inflammation of HepG2 cells induced by FFA.The results of immunohistochemistry,immunofluorescence and western blot showed that the expression of SREBP-1c,ACC,FAS and SCD-1 in vitro and in vivo was improved by RSK.The results showed that E Se could significantly improve the activation of NF-κB in NAFLD model in vitro and in vivo,and inhibit the translocation of p65 in cells.The results showed that the hypoglycemic effect of MT could be achieved by improving the sensitivity of insulin receptor.In addition,MT can improve NAFLD in type Ⅱ diabetes by improving glycogen synthesis,lipid synthesis,oxidative stress and inflammation in liver.MT can improve lipid accumulation of NAFLD model in vivo and in vitro by directly targeting SREBP-1c pathway in hepatocytes,and improve inflammatory response of NAFLD model in vivo and in vitro by directly targeting inhibition of NF-κB pathway activation in hepatocytes.In conclusion,MT may improve NAFLD in type Ⅱ diabetes based on the direct regulation of SREBP-1c and NF-κB pathway in hepatocytes.