Effect Of Low Temperature Stimulation On Gene Differential Expression In Adipocytes

Objective:The purpose of this study is to explore the differentially expressed genes of adipocytes at low temperatures.Methods:Primary ADSCs were successfully isolated from SD rats.Primary ADSCs began differentiation culture for 13 days in the third generation.The differentiated fat cells were identified by oil red O staining and divided into 37 °C control group(C group),30 °C(D1 group),20 °C(D2 group)and 10 °C(D3 group).After the four groups of adipocytes were cultured for 8 hours at different temperatures,the total RNA of each group of adipocytes was extracted for RNA-Seq detection and the differentially expressed genes were obtained by processing data by DEGseq software.Six differentially expressed genes were randomly selected to verify the accuracy of sequencing data by q RT-PCR.Using KOBAS3.0 software,STRING database and Cytoscape 3.5.0 software,GO gene functional enrichment analysis,KEGG biological pathway enrichment analysis and protein network interaction analysis were performed on differentially expressed genes.Results:ADSCs are successfully differentiated into adipocytes.Sequencing analysis showed that 35 of the differentially expressed genes in the CD1 group were up-regulated and 20 were down-regulated.Among the CD2 group,there were 121 differentially expressed genes,of which 76 were up-regulated and 45 were down-regulated.Among the CD3 group,there were 92 differentially expressed genes,of which 64 were up-regulated and 28 were down-regulated.GO analysis showed that differential gene function is mainly enriched in biological processes,and most of them are related to oxidative stress,angiogenesis and immune response.KEGG analysis revealed that the major enriched pathways of differentially expressed genes are PI3K-Akt signaling pathway,p53 signaling pathway,arrhythmogenic right ventricular cardiomyopathy,TNF signaling pathway,AGE-RAGE signaling pathway in diabetic complications,MAPK signaling pathway and mineral absorption.Differential expressed genes are associated with diabetes,myocardial disease,and inflammatory responses.Protein interaction network analysis showed that the key differential genes between group C,D1,D2 and D3 were Fos,Junb,Serpine1,Egr1,Csf1,Nr4a1,Myc,Cyr61,Rhob,Maoa and Gstp1.These genes are mainly in cell proliferation,neurodegeneration and Vascular regeneration and other aspects play a major role.Conclusion:Under low temperature stimulation,there are differential gene expression in adipocytes.The functions of differentially expressed genes are mainly concentrated in cell proliferation? immune inflammation?neuropathy and angiogenesis.Changes in the secretory function of adipocytes have an effect on adjacent tissues and organs.These provide new ideals for the pathogenesis of organs surrounding adipose tissue.