Study On The Differentiation Of PC12 Cells And Their Molecular Mechanism By Total Salvianolic Acid

Objective To study the differentiation of PC12 cells induced by Total salvianolic acid and its functional mechanism.Methods Establishing the differentiation model by PC12 cells,and observing the percentage of cell growth of PC12 cells in the induction of Total salvianolic acid.Detecting whether the different concentrations of Total salvianolic acid can peotect PC12 cell oxygen sugar deprivation(OGD).Observing the protein expression of PC12 cells after the ngc-2(MAP-2)monoclonal antibody is marked in the neuron specific marker microtubule-2(MAP-2).And using the immune imprinting method to detect the levels of ERK1/2,MEK1/2 protein and phosphorylation of ERK1/2,MEK1/2 protein.To more clearly define the role of ERK1/2,a small molecule inhibitor of MAPK/ ERK kinase named U0126 was examined for its effect on the cellular action of Total salvianolic acid 1μg·L-1 in PC12 cells.Total salvianolic acid 1μg·L-1 was selected for the subsequent differentiation experiment,and the expression of ERK1/2 and p-ERK1 /2 protein at different time points was detected by Western bloting method,which was used to detect the effects of Total salvianolic acid and NGF on PC12 cells.In the case of Total salvianolic acid 1μg·L-1,the PC12 cell1 h,4h,6h,8h,12 h,and 24 h were used to dye the MAP-2 and count the growth of PC12 cells.The expression of MEK1/2,p-MEK1/2 and Ras proteins in different time periods was detected by Western bloting method,and the expression of p-ERK1/2 and ERK1/2 protein was observed after the MEK inhibitor U0126 and PD184352 were used to perform PC12 cells.Statistical MEK inhibitor U0126 and PD184352 were used to determine the percentage of cell growth of the cell after the action of Total salvianolic acid 1μg·L-1 on PC12 cells.Results Total salvianolic acid 1μg·L-1,0.1μg·L-1,0.01μg·L-1 can all induce the growth of PC12 cells,among which,the function of 1μg·L-1 is most obvious.The survival rate of the PC12 cells is not affected by the different concentrations of Total salvianolic acid,and1μg·L-1,0.1μg·L-1 of Total salvianolic acid have repair effect on the PC12 cells after the OGD is damaged.The phosphorylation of MEK1/2 and ERK1/2 protein is associated with the dose concentration of salicylic acid.U0126 can reatrain the expression of p-ERK1/2.The cells appeared significantly in 1h,and the growth rate was unchanged at 4h,6h,8h,12 h and 24 h.The Western bloting test showed that the expression of p-ERK 1/2 was the most obvious after the NGF action cells,and the expression of p-ERK 1/2 protein was increased after 1h.p-MEK1/2,the expression of Ras protein also increased with time.MEK inhibitor U0126 and PD184352 inhibit protein activation(P<0.01).Conclusion(1)The results of MTT assay showed that Total salvianolic acid of1μg·L-1,0.1μg·L-1 and 0.01μg·L-1 was not toxic to the cells,and the total phenolic acid of1μg·L-1 had a proliferation effect on cells.(2)The Total salvianolic acid of 0.1μg·L-1 and1μg·L-1 has a restorative effect on OGD model.(3)Total salvianolic acid can induce the differentiation of PC12 cells which possibly by increasing the phosphorylation of ERK1/2.(4)After 1h,the effect of Total salvianolic acid action cells showed a growth phenomenon,and its differentiation was accomplished by activating the Ras/MEK1/2 / ERK1/2 pathway.