The Effect Of Adenovirus Type 36 On Browning Of 3T3-L1 Preadipocytes

Objective: To investigate the role of human adenovirus type 36(adenvirus type 36,Ad36)in inducing browning of 3T3-L1 preadipocytes.Methods:(1)BODIPY staining was performed on the 0,2,4,6 and 8 day cells of the MDI induction group(control group)and the MDI+Ad36 induction group(experimental group)to observe the cell fat formation.(2)The cells of the control group and the experimental group were collected on days 0,2,4,6,and 8 respectively.Real-time fluorescent quantitative PCR was used to detect the two brown fat marker genes pgc1α,ucp1,fgf21,mitochondrial oxidative phosphorylation related genes atp5 o,cox5b.The expression level of nd2 mRNA was changed;the protein expression level of the above two genes was detected by Western blot.(3)After the use of mTOR inhibitor rapamycin,BODIPY staining was used to detect cell fat formation;real-time quantitative PCR and Western blot were used to detect brown fat marker genes pgc1α,ucp1,fgf21,mitochondrial oxidative phosphorylation related genes atp5 o,Cox5b,nd2 mRNA and protein expression levels vary.Results:(1)During the differentiation of3T3-L1 preadipocytes,the lipid droplets in the MDI-induced group gradually increased and pooled,while the lipid droplets in the MDI+Ad36-induced group increased and pooled,on the 4th day of cell differentiation.After the addition of Ad36,the intracellular lipid droplet size gradually decreased compared with the control group.(2)The expression levels of pgc1α,ucp1,fgf21,atp5 o,cox5b,nd2 mRNA and protein were significantly higher in the experimental group than in the control group,and the difference was statistically significant(P<0.05).(3)After the use of mTOR inhibitor rapamycin,the ability of intracellular large lipid droplets to transform into small lipid droplets was significantly weakened;brown fat marker genes pgc1α,ucp1,fgf21,mitochondrial oxidative phosphorylation related genes atp5 o,cox5b,nd2 Both mRNA and protein expression levels were significantly decreased,and the differences were statistically significant(P<0.05).Conclusion: Ad36 may up-regulate the expression of pgc1α by mTOR,and further increase the gene expression levels of ucp1,fgf21,atp5 o,cox5b,nd2,accelerate the decomposition of triglyceride and mitochondrial oxidative phosphorylation,and finally make 3T3-L1 preadipocytes.Browning.