Research On The Effect Of Anshio â…¡ A And Ligustrazine To Induce BMSCs To Differentiate Into Myocardium-like Cells

Bone Mesenchumal Stem Cells(BMSCs) is a stem cell founded in bone marrows, where there is another one called hemopoietic stem cell. BMSCs are able to self-renew and self-duplicate, and they have multipotentiality. Because they are easy to collect, easy to separate, easy to be cultured and amplified (proliferate) in vitro, and low in immunogenicity, able to be divided into cardiac muscle cells, endothelial cells and vascular smooth muscle cells, they have become the most promising cellule de graine to treat the cardiac disorders. Therefore, the research on BMSCs'directional differentiation into myocardium-like cells under given conditions is of great importance and clinical significance in treatment of myocardial infarction with cellular transplantation. In recent years, it is reported in many literatures that active compounds in traditional chinese medicines can induce BMSCs to differentiate into cardiac muscle cells, especially the active ingredient (tanshioâ…¡a and ligustrazine) of the red sage root and astragalus membranaceus, two main compounds of Modified Red Sage Root Decoction.Purpose:To Probe into the effect of tanshioâ…¡a and ligustrazine, the two active compounded prescription (monomer) of Modified Red Sage Root Decoction, in extracorporeal co-inducement of BMSCs'directional differentiation into myocardium-like cells. Process and Methods:1.Separate, depurate and amplify BMSCs of Sprague Dawley rat (SD rat) in vitro and establish a matured and stable cultivation system of BMSCs.2.Measure the TDO of tanshioâ…¡a and ligustrazine respectively, the proportion of which can determine the prescription of co-inducement.3.Experiment of BMSCs inducement is divided into 5 groups, which are respectively with tanshioâ…¡a, with ligustrazine, with tanshioâ…¡a plus ligustrazine, with 5-aza and blank controller. Observe the expression of connexin43 after inducement with immunohistochemical method and supervise the changes in content of troponin with euzymelinked immunosorbent assay.Results:1.BMSCs from attachment method is high in purity, and the rate of their surface antigen CD11b and CD34 expression reached more than 98%, the rate of CD90 expression is less than 2%(as shown in the picture 1). As revealed from above, the cells are good in homogeneity, high in purity and well up to the lab requirements.2.According to the measurement of MMT chromometry, 0.01g/L is the TDO of tanshioâ…¡a and 0.039g/L of ligustrazine.3.There is no expression of connexin43 in blank controller after inducement, while in the other 4 groups there is weakly positive immune complex and positive cells in fusiform and polygonal form, which are colonially distributed. Optional density is high with 5-aza or tanshioâ…¡a plus ligustrazine than with tanshioâ…¡a or ligustrazine (P<0.05). whereas there is no statistical discrepancy between the former 2 groups.4.According to the measurement of troponin with enzyme linked immunosorbent assay, the changes in the content are different among the 4 groups and the group with tanshioâ…¡a plus ligustrazine is the highest.Conclusion:anshioâ…¡a plus ligustrazine can induce BMSCs to differentiate into cardiac muscle cells and then express connexin43 and troponin, of which the expressing strength is more than single ingredient.