Effects Of CYP450-EETs On Autophagy And Lipid Metabolism In Obese Mice

Objective: Epoxyeicosatrienoic acids(EETs)represent Arachidonic acid(AA)through Cytochrome P450 proteins,CYP450).EET analogues have some inhibitory effects on the early stages of adipogenesis.The aim of this study is to investigate the effect of CYP450-EETs pathway on the autophagy activity of adipose tissue in obese mice,and to observe the changes in body weight and lipid metabolism after exogenous EET treatment.To find new targets for the treatment of obesity and related metabolic diseases.Methods: Six weeks old mice were randomly divided into two groups: normal control group(n=10)and obesity group(n=40).The normal control group was given normal diet,and the obesity group was given high-fat diet.After 8 weeks of feeding,the body weight of the obese mice was 30% more than that of the control mice,and the obese mouse model was determined to be successfully established.After successful establishment of the obesity mouse model,the obesity group mice were randomly divided into obesity control group(OB group),EET group(11,12-EET treatment group),EEZE group(11,12-EET +11,12-EEZE treatment group).The EET group was given intraperitoneal injection of 11,12-EET,and the EEZE group was given intraperitoneal injection of 11,12-EET and 11,12-EEZE.After 14 days of treatment,mice were weighed and subcutaneous adipose tissue was collected.The body weight data of mice were analyzed to compare the differences in body weight of mice in each group.Western Blot was used to detect the expression levels of fatty acid synthase(FAS)and brown adipose tissue specific protein Uncoupling protein-1(UCP-1).The expression levels of autophagy marker proteins in subcutaneous adipose tissue of mice were detected.At the same time,the changes of the above indexes after treatment with exogenous EET(11,12-EET)and EETs antagonist(11,12-EEZE)were detected.Results: After 8 weeks of feeding with different diets in the normal control group and the obesity group,the average body weight of the obesity group was 30% higher than that of the NC group,suggesting that the obesity model was successfully constructed.After 14 days of different treatments,the EET group had a significant reduction in body weight compared with the OB group(P < 0.01),while there was no significant difference between the EEZE group and the OB group(P>0.05).Compared with the NC group,the expression of CYP2J2 protein in the OB group was significantly down-regulated(P <0.001),while the expression of CYP2J2 protein in the EET group was significantly lower than that in the EEZE group.Compared with the obesity group(OB),the EET treatment group had a significant reduction in FAS expression(P < 0.001),while the EEZE group had a significantly higher FAS expression than the EET group(P < 0.001).Compared with NC group,the expression level of UCP-1 in OB group was increased(P < 0.001).At the same time,by comparing the expression level of UCP-1 in the OB group and the EET group,we found that the expression level of UCP-1 was higher than that in the OB group(P< 0.05).The difference between EET group and EEZE group was statistically significant(P < 0.001).Compared with NC group,the protein expression levels of Atg6 and Becline-1 in OB group were significantly increased.Compared with the OB group,the EET group had significant increases in the protein expression levels of Atg6 and Becline-1(P < 0.01).The EEZE group had significantly lower protein expressions of Atg6 and Becline-1 than the EET group(P < 0.001).The detection of the expression level of P62 protein in each group showed that the expression level of P62 protein in the OB group was lower than that in the NC group(P < 0.01).Compared with the OB group,the expression level of P62 protein in the EET group was decreased(P < 0.001).Compared with the EEZE group,the EET group had a significant reduction in the expression of P62 protein(P < 0.001).Conclusions:CYP450-EETs pathway is an important factor affecting the development of obesity in mice,which may be achieved by regulating fat metabolism and adipose tissue Browning.The autophagic activity of adipocytes is up-regulated in obese mice,and exogenous EET can improve the autophagic activity of adipocytes in obese mice.Increasing the expression of CYP450-EETs pathway or exogenous application of EET can be used as a new method to reduce the incidence of obesity,inhibit fat accumulation,and promote the Browning of adipose tissue.