ABSTRACT
Objective@#To investigate the role of adenovirus type 36 (Ad36) in the browning of 3T3-L1 cells.@*Methods@#BODIPY staining was performed on the 0, 2nd, 4th, 6th and 8th days of the cocktail induction (control) group and the cocktail plus Ad36 induction (experimental) group to observe the adipogenesis of 3T3-L1 cells.The mRNA and protein expressions of uncoupling protein-1(Ucp1), ATP synthase, H+ transporting, mitochondrial F1 complex, O subunit (Atp5o), cytochrome c oxidase subunit 5B(Cox5b), and perilipin were detected by real-time PCR and Western-blot.@*Results@#The results of BODIPY staining showed that the lipid droplets in the control group gradually became larger with the differentiation of the cells, while the lipid droplets in the experimental group firstly became larger, and then appeared smaller after Ad36 was added on the fourth day. Compared with the control group, the mRNA and protein expression levels of Ucp1, Atp5o, and Cox5b in the experimental group were significantly increased while the expression level of perilipin was significantly decreased (all P<0.05).@*Conclusion@#During the differentiation of 3T3-L1 cells, Ad36 promotes its browning via increasing the expressions of Ucp1, Atp5o, and Cox5b.
ABSTRACT
Objective To investigate the role of LncRNA ROR in Ad36-induced browning of human adipose-derived stem cells(hADSC). Methods After hADSC was induced by cocktail and Ad36 for 2,4,6,and 8 days,Oil red O staining was performed for observing the adipogenic status. The mRNA expressions of LncRNA ROR, uncoupling protein 1(UCP1),and PRDM16 were detected by real-time PCR and the protein expressions of UCP1 and PRDM16 were detected by Western-blot. After LncRNA ROR was knocked down by siRNA, UCP1 and PRDM16 mRNA and protein expression levels in the process of Ad36-induced adipocyte differentiation were detected by real-time PCR and Western-blot. Results Oil red O staining showed that fat droplets in the cocktail-induced group were larger than those in the Ad36-induced group. Compared with the cocktail group,the mRNA expressions of LncRNA ROR, UCP1 and PRDM16, and the protein expressions of UCP1 and PRDM16 in Ad36 group were significantly increased(P<0.05). The mRNA and protein expressions of UCP1 and PRDM16 in LncRNA ROR knockdown group were significantly lower than those in control group(P<0.05). Conclusion In the process of Ad36-induced hADSC differentiation,the up-regulation of LncRNA ROR may stimulate UCP1 and PRDM16 expression,and thus promote the browning of hADSC.