Inhibition Of Akt2 Gene Expression Inhibits Differentiation Of Rabbit Preadipocytes

PI3K-AKT signaling pathway is one of the most insulin-sensitive signaling pathways.Previous studies have shown that PI3K-AKT signaling pathway may be involved in cell growth,development and differentiation.Many studies have shown that PI3K-AKT signaling pathway can regulate the adipogenic differentiation of some animal preadipocytes,but the role of PI3K-AKT in the adipogeneic differentiation of rabbit’s preadipocytes is still unclear.Therefore,the aim of this experiment was to explore the role of AKT2 gene in the differentiation of rabbit preadipocytes.The results are as follows:In this experiment,preadipocytes were successfully isolated from perirenal fat of newborn New Zealand rabbits by enzymatic digestion,and were cultured successfully.After cocktail-induction of adipogenic differentiation,oil red O staining was performed on 2d,5d and 8d.It was found that the red lipid droplets gradually increased and became larger with the differentiation.Meanwhile,the marker genes of adipocyte differentiation were identified by qRT-PCR.The level of PPARγ and C/EBPα at 2d increased significantly while decreased significantly at 5d and 8d.FABP4 was significantly increased dramatically on 2d and 6d.The expression of AKT2 gene in rabbit preadipocytes was identified by transfecting siRNA1,siRNA2 and siRNA3.The results showed that siRNA1,siRNA2 and siRNA3 could inhibit the expression of AKT2 gene in rabbit preadipocytes,and siRNA2 suppressed significantly AKT2 expression and the mRNA level was decreased by about 60%.Next,we inhibited the expression of endogenous AKT2 gene in rabbit preadipocytes via siRNA2,which inhibited the adipogenic differentiation of rabbit preadipocytes and down-regulated the synthesis of fatty acids.Besides,the content of TG as well as the expression of C/EBPα,PPARγ and FABP4 were significantly down-regulated.After transfected by the vector of pEGFP-AKT2,compared to control group and blank vector group,which promoted gene expression the adipogenic differentiation of rabbit preadipocytes and up-regulated the synthesis of fatty acids.Besides,the content of TG as well as the expression of C/EBPα,PPARγ and FABP4 were significantly up-regulated.To sum up,the model of rabbit preadipocyte induced differentiation in vitro was established successfully,and in the process of rabbit preadipocyte differentiation,inhibiting the expression of AKT2 gene can suppress the differentiation of rabbit preadipocyte.