| Objective: Based on previous proteomics studies,this article screens genes that are highly differentially expressed in the liver and associated with lipid metabolism.The effects of hypoxic environment,normoxic training,and hypoxic training on the expression of Fabp4,Acaa1 b,and Lyplal1 in the liver tissue of nutritionally obese rats and the secretion of serum exosomes were experimentally investigated."Signal" is activated by exosomes,explore the molecular mechanism of hypoxic training to induce exosome to regulate lipid metabolism,provide theory and experiments for improving the theory of hypoxic training to regulate lipid metabolism,and use non-medical methods to treat obesity and complications in accordance with.Methods: Seventy three-week-old female SD rats were established with a common diet control group(n=10,fed with ordinary feed for 8 weeks)and a nutritional obese rat model group(n=60,fed with high-fat diet for 8 weeks).Of the 51 nutritional obese rat models,40 rats were randomly selected and divided into 4 groups: the quiet control group(group C,n=10),the hypoxic quiet group(group H,n=10),often The aerobic training group(CE group,n=10 animals)and the hypoxic training group(HE group,n=10 animals)were treated for 4 weeks.Measure rat body weight,body length,and perirenal fat mass;and use serum lipid biochemical detection kits to detect TG,TC,LDL-C,and HDL-C levels in rat serum;use RT-PCR to detect Fabp4,Acaa1 b,Lyplal1 mRNA expression changes;WB was used to detect changes in Fabp4,Acaa1 b,and Lyplal1 protein expression in liver tissues;serum exosomes were finally extracted,and proteomics techniques were used to verify whether the "signals" of the lipid metabolism signal pathway were activated by exosomes.Result:(1)Compared with group C,Lee's index,body fat ratio,serum TC and TG in group H,CE and HE were significantly reduced(P<0.01);serum LDL-C in group H was significantly reduced(P<0.05),CE Serum LDL-C was significantly decreased in the group and HE group(P<0.01);HDL-C was significantly increased in the H group(P<0.05),and HDL-C was significantly increased in the CE group and the HE group(P<0.01);CE The weight of the group decreased significantly(P<0.05),and the weight of the HE group decreased significantly(P<0.01).(2)Compared with group C,Fabp4 and Acaa1 b mRNA and protein expressions in group H were significantly increased(P<0.05),Lyplal1 mRNA expression was significantly increased(P <0.05),Lyplal1 protein expression was significantly increased(P<0.01);CE group Fabp4 mRNA expression was significantly increased(P<0.05),Acaa1 b and Lyplal1 mRNA expression was significantly increased(P<0.01);Fabp4 and Acaa1 b protein expression was significantly increased(P<0.05),Lyplal1 protein expression was significantly increased High(P<0.01);Fabp4,Acaa1 b and Lyplal1 mRNA and protein expressions in HE group were significantly increased(P<0.01).(3)Compared with group H,TC and LDL-C in HE group were significantly decreased(P<0.05),Acaa1 b mRNA expression was significantly increased(P <0.05),HDL-C,Fabp4 mRNA and Acaa1 b protein expression were significantly increased(P <0.01);compared with the CE group,the body fat ratio and TC of the HE group were significantly reduced(P<0.05),Fabp4 and Acaa1 b mRNA expression were significantly increased(P<0.05),Acaa1 b protein expression was significantly increased(P<0.05).(4)TEM electron microscopy(200nm and 100nm)clearly observed the exosome extracellular vesicle-like structure in the four groups,and the exosomal WB verification was successful;the protein concentration measurement results showed that: hypoxic environment and hypoxic training It has a significant effect(P<0.05).(5)Label-free identified a total of 616 proteins,and there were 69 differential proteins between the 4 groups(P<0.05);comparison between the two groups found C and H,C and CE,C and HE,H and CE,H and There are 22,8,17,14,7,and 23 differential proteins between HE,CE,and HE respectively.There are 11,7,10,12,5,and 8 up-regulated proteins between groups,and 11 down-regulated proteins,respectively.,1,7,2,2 and 15.(6)Significantly enriched GO analysis between the two groups found that GO terms related to lipid metabolism between C and H,C and CE,C and HE,H and CE,H and HE,and CE and HE were 2,8,and 7,respectively.,4,13,and 9.(7)Significant enrichment between the two groups of KEGG analysis found that there are 7 significant pathways related to lipid metabolism,namely: Galactose metabolism(key proteins: AKR1 B and PFKA),Fat digestion and absorption(key proteins: APOA-1 and APOB-48),Cholesterol metabolism(key proteins: APOA-1,APOB-100 and PCSK9),Glycerolipid metabolism(key proteins: ALDH,AKR1 B and LPL),Starch and sucrose metabolism(key proteins: PYG and GLGP),PI3K-Akt signaling pathway(Key proteins: VWF,CD41,Thbs1,MSFL,and KCIP-1)and Fructose and mannose metabolism(key proteins: AKR1 B,PFKA,and ALDO).Conclusion:(1)Hypoxic environment,normoxic training and hypoxic training can significantly increase the expression of Fabp4,Acaa1 b and Lyplal1 mRNA and protein,indicating that RT-PCR and Western Blot verification are consistent with the previous proteomics results,and the effect of hypoxic training is relatively Good;hypoxia training promotes the expression of Fabp4,Acaa1 b and Lyplal1 proteins in the liver of nutritionally obese rats,thereby promoting fatty acid oxidation and ultimately improving lipid metabolism.(2)After three different intervention methods,it was found that there are 7 signal pathways in which serum exosomes of nutritionally obese rats are significantly related to lipid metabolism,of which hypoxia training significantly interferes with 4 pathways: fat digestion and absorption pathway(key protein: VWF,CD41,Thbs1,MSFL and KCIP-1),cholesterol metabolism pathway(key proteins: AKR1 B,PFKA and ALDO),PI3K-Akt signaling pathway(key proteins: APOA-1 and APOB-48)and metabolism of fructose and mannose The pathways(key proteins: APOA-1,APOB-100,and PCSK9)indicate that some "signals" about lipid metabolism signaling pathways are indeed activated by exosomes. |
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