Effective Component Of Salvia Miltiorrhiza And Relevant Mechanism In Promoting Cardiomyogenic Differentiation Of Human Placent-derivedmesechymal Stem Cells

ObjectiveTo assess theeffective component of Salvia miltiorrhiza in promoting cardiomyogenic differentiation of human placent-derivedmesechymal stem cells and investigate the relevant mechanism of it.Methods1.h PDMSCs were isolated by tissue culture which was identified by flow cytometer.2.To intervent the 4 to 6 generation cells with different concentrations of Salvia miltiorrhiza,Danshensu,Salvianolic acid B,Protocatechuic aldehyde,Tanshinone ?,Tanshinone?A,Cryptotanshinone.MTS assay were done to analyze the effect of above-mentioned drugs in cells proliferation,then choose a non-toxic dose(without an affected critical concentration)that has no effect on cell proliferation to experimentize.3.To intervent the h PDMSCs with the above-mentioned drugs and install the control group at the same time.We Observate the cells morphological changes under microscope every day and change the liquid every 4 days.On the 20 th day,tested the expression of a-sarcomeric actin(?-SCA)in each group by immunohistochemical assay and the expression of GATA4,atrial natriuretic factor(ANF)and cardiac troponin-I(c Tn I)by Western-blot assay.Detected the expression of c Tn I by immunofluorescence and counted the cardiomyocyte differentiation rate.4.To test the expression of GSK-3? and ?-catenin by Western-blot assay.Results1.h PDMSCs was successfully identified by tissue culture.2.The concentration of Salvia miltiorrhiza(? 4mg/L),Danshensu(?10mg/L),Salvianolic acid B(?10mg/L),Protocatechuic aldehyde(?0.5mg/L),Tanshinone?(?1mg/L),Tanshinone?A(?0.1mg/L)and Cryptotanshinone(?0.1mg/L)can reduce the proliferation rate of cells in different degrees.In some cells,the volume increased,changed to polygon with only one nucleus,and was similar to the cardiac myocytes.The expression of cardiac specific protein(?-SCA,GATA4,ANF,c Tn I)can be promoted in different degrees,the effects of Tanshinone?A are the most obvious comparing with the groups which are mentioned above.3.Salvia miltiorrhiza and its effective component can increase the expression of GSK-3? and decrease the expression of ?-catenin.ConclusionsThe most effective component of Salvia miltiorrhiza in promoting cardiomyogenic differentiation of human placent-derivedmesechymal stem cell is Tanshinone ?A.The mechanism is related with the Wnt/?-catenin signal pathway.