Herba Epimedii Effects On Differentiation Of Bone Cells And On Expressions Of CD44 And FN

ObjectiveTo determine the effects of Herba Epimedii on formations of osteoblasts and osteoclasts in vitro , and to study the expressions and effects of CD44 and FN in bone cells. To clear the molecule mechanism in curing osteoporosis by Herba Epimedii .MethodPhase One: Bone marrow mesenchymal stem cell culture systems were divided into three groups, designated: M, D and C Groups. Herba Epimedii serum was added to M Group; Dexamethason was added to D Group; and C Group was maintained as the control group. On the second, fifth and eighth days of the experiment, Giemsa staining and immunocytochemistry were used to observe the shape of the cells and the expression of CD44 and FN. ARS staining was also used to detect mineralized nodes on the eighth day. Results of the staining were analyzed using IPP image analysis software and SPSS13.0.Phase TwoBone marrow stem cell culture systems were cultured with RANKL and M-CSF and divided into three groups, designated: M, A and C Groups. Herba Epimedii serum was added to M Group; antibodies of CD44 were added to A Group; and C Group was maintained as the control group. On the third, sixth, and ninth days, Giemsa staining, tartaric-resistance and immunocytochemistry were used to observe the shape of the cells, osteoclasts and expression of FN. Results of the staining results were analyzed using IPP6.0 image analysis software and SPSS13.0.ResultsPhase One: Many mineralized nodes were found in M and D Groups, with M Group having a larger number of nodes than D Group. Few nodes were found in C Group. There was no apparent difference in CD44 expression among all three groups. Expression of FN in M and D Groups was significantly less than in C Group. On the fifth and eighth days, FN expression increased in M and D Groups and exceeded that in C Group.Phase Two: All three groups had osteoclast-like cells, but the percentage of TRAP+ cells was significantly different. M Group was determined to have lowest percentage of TRAP+ cells. C Group had the highest percentage. The cells in all three groups expressed FN, but expression of FN in mononuclear macrophages was greater than in multinuclear cells.Conclusion:Herba Epimedii will not only induce marrow mesenchymal stem cells to differentiate into osteoblasts, but also inhibite the differentiation of osteoclast from bone marrow cells. Herba Epimedii was found to have an effect on the formation and apoptosis of osteoclasts by inhibiting the expression of CD44 in osteoblasts. FN is also an important factor in the course of Herba Epimedii regulating the bone formation and bone resorption .