| Purpose:Exercise is an effective means to regulate the energy imbalance of the body induced by high-fat diet.PPARγ is a transcription factor.It plays an important role in the browning of white fat,promoting the differentiation of adipocytes,regulating the lipid metabolism of adipose tissue,improving insulin resistance and inhibiting inflammation by affecting downstream target genes.In this study,the methods of co-immunoprecipitation(CO-IP)and mass spectrometry were used to analyze the proteomics of PPARy and its interacting proteins.Based on the known pathways,this study explored the effects of aerobic exercise on the PPARγ protein of high-fat diet rats.The influence of interacting proteins and possible pathways.Methods:A total of 32 SPF male SD rats aged 3 weeks.After the rats were adaptively fed for one week,they were randomly divided into 4 groups,namely the standard diet quiet group(group C),the standard diet aerobic exercise group(CE group),the high-fat diet quiet group(H group),and the high-fat diet aerobic exercise Group(HE group),8 in each group.Rats in each group were subjected to 8 weeks of experimental intervention.Among them,the quiet group was raised quietly,and the exercise group was subjected to an 8-week aerobic exercise intervention program.Exercise 5 days a week,each intervention for a total of 60 minutes,including 5 minutes of warm-up phase and 50 minutes of training Phase,5min recovery phase.The standard diet group was fed with standard feed,and the high-fat diet group was fed with highfat feed.48 hours after the last exercise of the rats in the exercise group,the rats were anesthetized by intraperitoneal injection,the rats were weighed,and the body fat percentage of the rats was measured with a DEXA meter.After blood was taken from the abdominal aorta,the subcutaneous white fat in the groin was quickly separated and clipped,and the immunoprecipitation experiment,mass spectrometry analysis,differential protein screening and bioinformatics analysis were performed.Results:(1)After 8 weeks of intervention,the weight of rats in each group increased significantly.Compared with group C,the body weight and body fat rate of rats in CE group were significantly reduced(P<0.01);compared with group H,the body fat rate of rats in HE group was significantly reduced(P<0.05);compared with group CE,The weight of rats in the HE group increased significantly(P<0.01),and the body fat rate also increased significantly(P<0.05).(2)In rat white fat,compared with group C,the protein expression of PPARy in group H was significantly reduced(P<0.05);the total number of differential proteins was 33,1 1 were up-regulated and 22 were downregulated.Compared with the CE group,the protein expression of PPARγ in the HE group showed an upward trend;the total number of differential proteins was 19,5 were up-regulated,and 14 were down-regulated.The 8-week high-fat diet intervention mainly affects signaling pathways such as cholesterol binding,low-density lipoprotein receptor binding,and adrenaline signaling by affecting PPARy and its interacting proteins,thereby participating in the negative regulation of lipid storage.(3)Compared with group C,the protein expression of PPARy in CE group showed a downward trend;the total number of differential proteins was 50,12 were up-regulated,and 38 were down-regulated.Compared with the H group,the protein expression of PPARy in the HE group was significantly increased(P<0.05);the total number of differential proteins was 25,6 were up-regulated.and 19 were down-regulated.8-week aerobic exercise intervention mainly affects obesity-related signaling pathways such as drug metabolism,diabetic complications,and aldosterone-regulated sodium reabsorption by affecting PPARγ and its interacting proteins.It also affects cholesterol binding,lowdensity lipoprotein receptor binding;fibroblast growth factor,angiogenic factor production;phospholipase activity,glycophospholipid metabolism process,and then affects lipid metabolism process.Conclusions:(1)The 8-week high-fat diet may inhibit the body’s inflammatory response by down-regulating the expression of PPARy protein and its direct interaction protein Crp,and affect the body’s cholesterol binding and low-density lipoprotein receptor binding,and regulate lipid metabolism.(2)Under the intervention of 8 weeks of aerobic exercise,PPARγ protein may directly down-regulate the Crp protein,and at the same time,it may affect the drug metabolism,diabetes and other related pathways by down-regulating the expression of Collal protein and Prdx6 protein,and reduce lipid storage,thereby affecting The process of glucose and lipid metabolism.(3)In the 8-week high-fat diet rat model under the intervention of aerobic exercise,although no protein that directly interacts with PPARy was found,the differential protein function is enriched in redox homeostasis,diabetes,amino acid metabolism,etc.,and the body’s The energy metabolism process is closely related. |
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