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Abstract To clarify the function of miR-128a during the differentiation of rat (Rattus norvegicus) preadipocytes, we detected the expression level of miR-128a during adipogenesis and clarified its expression tendency during differentiation; then we constructed an over-expression adenovirus vetor of pre-miR-128a and infected rat preadipocytes. Subsequently, the expression of adipogenesis marker peroxisome proliferator-activated receptor gamma(PPARγ) and adipocyte protein 2(aP2) were analysed by Real-time PCR and Western blot in mRNA and protein level, respectively. After the adenovirus vector infecting rat preadipocytes, we detected lipids accumulation in 10th day by Oil-red O staining, observing the adipogenesis in morphology. Results showed that the expression level of miR-128a was at the minimum point in the next day of the adipocyte differentiation, and then maintained at the level of day 0. After the over-expression of miR-128a, the expression levels of adipocyte marker genes PPARγ, aP2 and the number of lipid droplets had obviously declined, compared with that in control. The results showed that miR-128a could inhibit the differentiation of pre-adipocytes in rat. We suspected that PPARγ is a potential target gene of miR-128a according to the changes of mRNA and protein expressing levels observed after the treatment of adenovirus infection in rat pre-adipocytes. Also, this study supplies the basic theory for the further research about the function of miR-128a during the differentiation of adipocytes.
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Received: 25 October 2012
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