| Objective:In view of the fact that the mechanism of the effect of HIIT on fat metabolism has not been clarified in detail,and PPAR γ is a key factor in regulating fat formation and energy metabolism.In this study,Co-immunoprecipitation combined with mass spectrometry was used to explore the potential physiological function and metabolic pathway of PPARγ in white adipose tissue,the effect of high-fat diet on PPAR γ network in white adipose tissue,and the PPAR y-mediated mechanism of HIIT regulating lipid metabolism.Methods:32 SD male rats(3 weeks old)were randomly divided into 4 groups:standard diet quiet group(CS group),high fat diet quiet group(HS group),standard diet exercise group(CE group)and high fat diet exercise group(HE group),with 8 rats in each group.The rats in each group were intervened with diet and exercise for 8 weeks.The rats in the standard diet group were fed with standard diet(D12450B),and the rats in the high-fat diet group were fed with high-fat diet(D12451).The rats in the exercise group were given high-intensity intermittent exercise for 8 weeks,alternately with the exercise intensity of 70%V O2max,90%V O2max and 50%VO2max on the treadmill,5 times a week,each time 50min.48 hours after the last exercise,the rats were weighed and killed after DEXA body composition test,and the subcutaneous white adipose tissue in the groin was quickly separated and cut.Three rats in each group were randomly selected for immunoprecipitation and mass spectrometry analysis.The differential proteins were screened by 1.5 times of the difference multiple and P-value 0.05 as the standard,and then bioinformatics analysis was carried out.Results:(1)compared with CS group,the body fat percentage of HS group was significantly higher(P<0.05),lean body weight was significantly lower(P<0.05),and the expression of PPAR γ in white adipose tissue was significantly lower than that of HS group(P<0.05).Fifty differential proteins were screened,including 3 up-regulated proteins and 47 down-regulated proteins,which were mainly enriched in antigen-antibody binding,immune response,glucose metabolism and other signal pathways.CRP is a protein that directly interacts with PPAR γ and is down-regulated.(2)compared with CS group,the body weight of CE group decreased significantly(P<0.05),and the expression of PPAR γ in white adipose tissue had no significant difference(P>0.05).Forty-five differential proteins were screened,including 26 up-regulated differential proteins and 19 down-regulated differential proteins,which were mainly enriched in protein binding,glucose metabolism,virus infection and other signal pathways.CRP is a protein that directly interacts with PPAR γ and is down-regulated.(3)there was no significant difference in the expression of PPAR γ in white adipose tissue between HE group and CE group(P>0.05).Fifty-nine differential proteins were screened,including 14 up-regulated differential proteins and 45 down-regulated differential proteins,which were mainly enriched in protein binding,enzyme activity regulation,immune response,glucose and lipid metabolism and other signal pathways.Fasn is a protein that directly interacts with PPAR γ and is down-regulated.(4)compared with HS group,the body fat percentage of HE group was significantly lower(P<0.05),lean body weight was significantly higher(P<0.05),and the expression of PPAR γ in white adipose tissue was significantly higher than that of HE group(P<0.05).51 differential proteins were screened,including 36 up-regulated differential proteins and 15 down-regulated differential proteins.It is mainly enriched in signal pathways such as enzyme activity regulation and glucose metabolism,and there is no direct interaction with PPAR γ.Conclusion:(1)High-fat diet for 8 weeks can down-regulate the expression of PPAR γ in subcutaneous white adipose tissue.PPAR γ may participate in the process of antigen-antibody binding,immune reaction and glucose metabolism through potential interaction proteins.(2)8-week high-intensity intermittent exercise did not change the expression of PPAR γ in subcutaneous white adipose tissue,but it may improve the pathways of protein binding,glucose metabolism and virus infection through potential interaction proteins.(3)under the high-fat diet model,8-week high-intensity intermittent exercise up-regulated the expression level of PPAR γ in subcutaneous white adipose tissue,and PPAR γ may be involved in the processes of enzyme activity,protein binding,immune reaction,glucose and lipid metabolism through potential interaction proteins. |
PDF Full Text Request