Effect Of DLGAP1 Gene On White Adipose Browning In Mice

Obesity is one of the main reasons that endanger people’s health nowadays.Obesity leads to a large amount of fat synthesis and the metabolism slowly in the body,which makes people vulnerable to chronic metabolic diseases.White adipose tissue stored in mammals can be converted into energy-consuming brown adipose tissue under certain conditions to enhance fat metabolism and alleviate obesity symptoms,but the mechanism of browning of white adipose tissue is still unclear.In order to understand the effect of DLGAP1 on browning of white adipose tissue,we investigated the role of DLGAP1 in browning of white adipose tissue by overexpression or knockout of DLGAP1 in adipose.In this study,the expression of DLGAP1 in white adipose tissue and brown adipose tissue of mice was detected.It was found that the expression of DLGAP1 in white adipose tissue was significantly higher than that on brown adipose tissue.The changes of related gene expression and the effects of DLGAP1 overexpression on proliferation,apoptosis and lipid induction in brown adipose was detected by q-PCR,Western Blot,immunofluorescence and other biological techniques.The results showed that overexpression of DLGAP1 in brown adipose increased the expression of white adipose tissue marker genes ASC1,LEP,FASN and SCD,but the expression were decreased of brown adipose tissue marker genes UCP1,ADRA1 A,PGC1ɑ and CIDEA.Overexpression of DLGAP1 can inhibit the proliferation and the formation of small lipid droplets in brown adipose,promote and apoptosis of brown adipose.Finally,the DLGAP1 CRISPR/Cas9 knockout vector was constructed and transfected into white adipose.The changes of gene expression and the effects of DLGAP1 knockout on adipocyte proliferation,apoptosis and lipid induction in white adipose was detected by q-PCR,Western Blot,immunofluorescence and other biological techniques.The results showed that DLGAP1 knockout decreased the expression of ASC1,LEP,FASN and SCD in white adipose;and increased the expression of UCP1,ADRA1 A,PGC1ɑ and CIDEA in white adipose.DLGAP1 knockout inhibits the apoptosis of white adipose,and promotes white adipose to form aggregated lipid droplets with brown adipocyte characteristics.Our study found that the knockout of DLGAP1 in white adipose had no singnification effect on proliferation.These results confirmed that DLGAP1 could negatively regulate the browning of white adipose tissue in mice.