The Research Of C2C12 Myogenesis Mediated By Adipose And Regulation Of TNF-α

The quality of meat and diseases such as heart disease, muscle atrophy are related to muscle differentiation. Adipose (adp) encodes an evolutionarily conserved and broad spectrum protein. Adp was originally discovered in the obesity Drosophila mutant. Adp mutant flies can efficiently store fat and survive in a starvation survival assay. But it is kown poorly about the function now. Using C2C12 myoblasts, we investigated the role of ADP and TNF-α, an adipocytokin during C2C12 myogenesis by building stable cell lines expressing ADP, RT-PCR, Western Blot, immunofluorescent staining.The detailed results are as follows:1. The expression of ADP increased during C2C12 myogenesis.2. Stable cell lines expression ADP has a higher differentiation efficiency in advance comparing with control. Also MHC, the myogenic maker showed higher expression comparing with control during myogenensis. These proves that ADP can promote C2C12 differentiation.3. Analysis the expression of Myf5, MyoD1, MyoG, Mef2c in stability transfected cell lines during myogenensis, we find that only MyoG level was increased. While overexpression of ADP or knocked down ADP in C2C12 telled the expression of MyoG varied with ADP. Thesis showed that ADP promotes myogenesis maybe by regulating MyoG.4. Analysis was taken 2 days after TNF-α stimulating stable transfection cell lines during myogenesis. MHC, ADP and KLF15 were effectively decreased with TNF-α. And immunofluorescent staining showed the quatity of myotubes was significantly reduced. These proved ADP can promote C2C12 differentiation and TNF-α can inhibit it.