| With people' living standard improving, people request quality of meat and preparation better. The type of skeletal muscle fiber and ratio are important for meat quality. But we can not understand completely development process of skeletal muscle and regulation mechanism. Generally we think muscular histogenesis has many steps of biology, including foetal mesoderm merotomy; myoblast shapes in arthromera; myoblast can form muscular tissue to move to limbs; the proliferation of sarcoblast can increase cells'quatity.In certain process of development, sarcoblast exits cell cycle and differentiates muscle fiber expressing specific gene of skeletal muscle. People have had study to indicate insulin and insulin-like growth factor play an important part in skeletal muscle development. Insulin is a multifunctional hormone proteinoid, it participates metabolic regulation of organism, and binds special receptor of cell surface to promote the proliferation of cells. Insulin-like growth factor is also a multifunctional regulating factor on the proliferation of cells, it promotes the proliferation and differentiation of cells, and growth and development of organism. People have studied the relation of skeletal muscle and IGF-1 systematically and penetratingly, but they have no much about the relation of skeletal muscle and insulin. Because of regulating insulin signal important in medical, more and more material regulating insulin signal are found by people. For example ursolic acid, sodium vandate and AG1024.We study the effect of these material on the proliferation and differentiation of skeletal muscle cells to elucidate the significance of insulin signal pathway in skeletal muscle development, and provide rationale of improving meat quality by regulating insulin signal. Material and method:①Proliferation of myblast C2C12:We manipulate C2C12 by insulin,UrsoIic acid, Sodium vandate and AG1024. The concentrations of insulin are 0,10-9mol/L, 10-8mol/L, 10-7mol/L; the concentrations of Sodium vandate are 0,10μmol/L,25μmol/L,50μmol/L; the concentrations of AG1024 are 0,1μmol/L, 10μmol/L, 100μmol/L; the concentrations of ursolic acid are 0, 10-7mol/L,10-8mol/L,10-9mol/L. Then we measure MTT value to reflect cells proliferation every 24h, successively three days.②Differentiation of myblast C2C12: We manipulate C2C12 by insulin,Ursolic acid, Sodium vandate and AG1024 for 4days. The concentrations of insulin are 0,10-9 mol/L, 10-8 mol/L,10-7 mol/L; the concentrations of sodium vandate are 0, 0.5μmol/L,2.5μmol/L,5μmol/L;the concentrations of AG1024 are 0,0.001μmol/L, 0.01μmol/L,0.1μmol/L,1μmol/L; the concentrations of ursolic acid are 0,10-7mol/L, 10-8mol/L, 10-9mol/L. then we calculate CK value and Giemsa dyeing to observe cell picture to detect differentiation of cells.Result: insulin promoted to proliferate and differentiate on C2C12, when the concentrations of insulin was 10-8mol/L, effect of proliferation on C2C12 was best, when the concentrations of insulin was10-7mol/L, C2C12 differentiate best; effect of sodium vandate on proliferation of C2C12 was close with concentration and time, if the concentration was over10uM,and time is excess24h, sodium vandate inhibited cells to proliferate evidently, it has no effect on differentiate;AG1024 inhibited cells to proliferate evidently, but when the concentration of AG1024 is in 0.1～0.001μmol/L,it promoted C2C12 to differentiate, and when we had 0.01μmol/L AG1024, effect of differentiation was best; ursolic acid inhibited C2C12 to proliferate with concentration and time closely, and it had no effect on differentiation of C2C12.Conclude: Insulin can promote C2C12 to proliferate and differentiate at the certain concentration; sodium vandate can promote proliferation of C2C12,but have no effect on differentiation;AG1024 inhibits C2C12 to proliferate, but it can promote differentiation of C2C12 at certain concentration.; ursolic acid inhibits C2C12 to proliferate and has no effect on differentiation of C2C12.
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