The Role And Mechanism Of Adiponectin As LncRNA In Preadipocytes Diffrentiation

Long noncoding RNAs(lncRNAs)are a kind of noncoding RNA,which have a key role during gene expression,and can take part in diverse biological processes.Adiponectin,a typical secreted factor of adipose,plays an essential function during differentiation processes of preadipocytes and in normal metabolism,and it is under the control of transcriptional and post-transcriptional levels.Here,we predicted a long noncoding RNA,Adiponectin AS lncRNA,transcribed from the opposite strand of adiponectin,and explored the role and mechanism of it on preadipocytes differentiation and its impact on mouse metabolism.In this study,we used biological softwares to analyse the coding potential of Adiponectin AS lncRNA;RT-PCR was conducted to identify its transcription in adipose.Tissues profile,differentiation expression profile and stablity of Adiponectin AS lncRNA were detected by RT-qPCR.FISH detected its localization in cells.Recombinant adenovirus were constructed to infect mouse primary preadipocytes.RT-qPCR,WB and Oil red O staining were used to research the function of Adiponectin AS lncRNA on preadipocytes differentiation,and the mechanism were explained by RPA.Finally,recombinant adenovirus were injected into mouse,GTT,ITT,ELISA,RT-qPCR and TG kit were used to detect its role on mouse lipid metabolism.The main findings are as follows:1.Adiponectin AS lncRNA widely expressed in heart,liver,spleen,brain,kidney,muscle and adipose,with higher expression level in adipose,liver,and spleen;during the differentiation processes of 3T3-L1 cell,Adiponectin AS lncRNA expression level was increased greatly2.Adiponectin AS lncRNA was positioned on both nucleus and cytoplasm in preadipocytes cell line,but with more fluorescence signal in cytoplasm in mature adipocytes.3.Overexpression of Adiponectin AS lncRNA will decrease level of PPARγ、adiponectin and aP2 mRNAs and proteins significantly(P < 0.05);Adiponectin AS lncRNA binds with adiponectin to form RNA duplex to inhibit translation of adiponectin;4.In HFD group,total length and non-overlapping recombinant adenovirus decreased mouse food intake,and the weight of adiposes and liver was lower than control group(P <0.05).The glucose tolerance and insulin sensitivity were better in non-overlapping group thanin control and total length group in HFD condition.There were higher TG content in mouse liver,muscle and serum(P < 0.05)in control group compared with treatment groups,and the serum adiponectin level had no difference between control and treatment groups,but there was a significantly decrease of leptin in both treatment groups(P < 0.05).Collectively,Adiponectin AS lncRNA can form RNA duplex with adiponectin mRNA to prevent its translation,and inhibit the differentiation of mouse preadipocytes.Overexpression of Adiponectin AS lncRNA will inhibit mice adipose tissues weight,decrease liver TG content,promote insulin signaling pathway relative genes expression and improve glucose tolerance and insulin sensitivity.These results provide a target gene for adipogenesis,and also provide a scientific basis for meat quality traits in breeding.