Effects Of Entacapone Combined With Exercise On The Expression Of FTO On Glucose And Lipid Metabolism In Obese Mice

With the rapid development of society and the continuous improvement of human economic living standards,the number of obese and overweight people is increasing.Obesity has become one of the serious health problems that people have to face and solve.Exercise is recognized as one of the safe and effective ways to lose weight,and it is also important to find safe and effective drugs to treat obesity.FTO is a gene related to obesity.Many studies have found that FTO single nucleotide polymorphism is highly associated with obesity.FTO participates in glucose and lipid metabolism and energy metabolism and plays an important role.Entacapone is a specific inhibitor of FTO gene,which can inhibit the expression of FTO.Related studies have also shown that exercise can regulate the expression of FTO.In this study,the effects of intragastric administration of Entacapone,exercise and Entacapone combined exercise on liver metabolism and hypothalamic energy metabolism in obese Kunming mice were studied,and the effect of drug combined with exercise intervention on obesity was evaluated.The main research contents and results are as follows:（1）Study on the physiological and biochemical effects of Entacarbon combined exercise on obese mice.Obese mice were induced by high fat diet and divided into obesity group（OB）,Entacarbon group（ENT）,exercise group（EX）and Entacarbon combined exercise group（ENT-EX）,and normal control group（CK）.The experiment period was 5 weeks.The body weight,body fat,blood lipid and blood glucose,serum AST,ALT,liver TG and TC levels of mice were detected,and the morphological changes of liver tissues were observed.The results showed that the weight and body fat content of obese mice decreased significantly after entacapone combined with exercise intervention.Serum TG,TC and LDL-C levels were significantly decreased,while HDL-C levels were significantly increased.Blood sugar levels dropped significantly;Serum AST/ALT levels were significantly reduced.After entacapone combined exercise intervention,the levels of TG and TC in liver decreased significantly.Liver HE sections showed severe liver tissue lesions in the OB group,which was significantly improved in the ENT group,ENT-EX group and EX group.The results showed that entacapone combined with exercise intervention had significant effect on weight loss and lipid reduction in obese mice,and improved lipid level and lipid accumulation in liver,and had a certain protective effect on liver injury.（2）Effects of entacapone combined with exercise on glucose and lipid metabolism in liver of obese mice,and mRNA and protein expression levels of FTO,AMPK and glucose and lipid metabolization-related genes in liver were detected by fluorescence quantitative PCR and western blot.The results showed that the mRNA and protein expression of FTO gene in liver were significantly decreased after entacapone combined exercise intervention.The expression levels of AMPK and P-AMPK protein in liver were significantly increased.The mRNA and protein expressions of Fas,Sreb P-1C,CD36 and ACC genes related to liver lipid synthesis were significantly decreased.The mRNA expressions of ACO,CPT-1 and PPARα genes related to fatty acid oxidation in liver were significantly increased.ACO protein expression was significantly increased in EX group,CPT-1 protein expression was significantly increased in ENT group and ENT-EX group,while PPARα protein expression had no significant difference among all groups.The mRNA and protein expressions of gluconeogenesis related gene G6P and PEPCK gene in liver of obese mice were significantly decreased after entacapone combined with exercise intervention.The mRNA and protein expression of liver glucose transporter GLUT4 gene were significantly increased.The results showed that entacapone combined with exercise intervention may activate AMPK pathway by inhibiting FTO expression,and further affect the expression of liver glucose and lipid metabolism genes to regulate liver metabolism.（3）Preliminary study on the effects of entacapone combined exercise on hypothalamus FTO expression and m6A methylation in obese mice.RNA m6A methylation kit was used to detect hypothalamus m6A methylation level,and fluorescence quantitative PCR and western blot were used to detect hypothalamus FTO expression.The results showed that the m6A methylation level of hypothalamus of mice was basically not affected by entacapone combined with exercise intervention,and there was no significant difference among groups.The mRNA and protein expression of FTO gene in hypothalamus were significantly decreased.The results suggest that entacapone and exercise intervention may reduce body weight in obese mice by inhibiting hypothalamus FTO expression.In conclusion,entacapone combined with exercise intervention can inhibit the expression of FTO in the liver of obese mice,activate AMPK pathway,regulate the expression of genes related to liver glucose and lipid metabolism,and down-regulate the expression of FTO in the hypothalamus of mice,ultimately achieving the effect of improving the disorder of liver glucose and lipid metabolism,and reducing the weight and fat content of obese mice.This study lays a foundation for the improvement effect of drugs combined with exercise intervention on obesity,and provides theoretical and experimental basis for the treatment of obesity and the development of functional drugs.