Experimental Study On The Effects Of Different Concentrations Of Icariin On Osteogenic Differentiation And RunX2, OSX And ?-catenin Of Rat Tendon Stem Cells

Objective:To investigate the effects of different concentrations of Icariin on osteogenic differentiation of rat tendon stem cells(TDSCs),Runx2,OSX m RNA?-The effect of catenin protein expression provides a theoretical basis for the application of icariin in promoting tendon bone healing.Methods:TDSCs of SD rats were cultured and subcultured to 3 generations for standby.Different concentrations of icariin were selected to act on cells,and the effects of different concentrations of Icariin on cell activity were observed by MTT method.The concentration interval with the least influence(10^-9-10^-5mol/L)was selected.Rats were treated with icariin 0mol/L Group(control group),icariin 10^-9mol/l group,icariin 10^-8mol/l group,icariin 10^-7mol/l group,icariin 10^-6mol/l group and icariin 10^-5mol/l group.Rats were treated with icariin combined with osteogenic differentiation induction medium for 21d.ALP staining was used to observe ALP precipitation,Alizarin red staining was used to observe the distribution of calcium nodules,RT-PCR was used to detect the expression of Runx2and OSX related genes,and Western blot was used to detect them?-Expression of catenin protein.Results:(1)under microscope,ALP staining showed blue sediment aggregation,alizarin red staining showed a large number of calcium nodules,which were positively correlated with the concentration of icariin.(2)RT-PCR was used to detect the expression of Runx2 and OSX m RNA.Compared with the control group,the expression of Runx2 increased in 10^-7mol/l group,10^-6mol/l group and 10^-5mol/L Group(P<0.05),especially in 10^-6mol/l group and 10^-5mol/L Group(P<0.01);There was no significant difference in the expression of Runx2 between 10^-9mol/l group and10^-8mol/l Group(P>0.05).Compared with the control group,the expression of OSX m RNA in 10^-8mol/l group,10^-7mol/l group,10^-6mol/l group and 10^-5mol/l group increased significantly(P<0.05),especially in 10^-6mol/l group and 10^-5mol/l Group(P<0.01);There was no significant difference in the expression of 10^-9mol/l Group(P>0.05).(3)Western blot detection?-Expression of catenin protein.Compared with the control group,10^-8mol/l group,10^-7mol/l group,10^-6mol/l group and 10^-5mol/l Group?-The expression of catenin protein increased significantly(P<0.01).Conclusion:(1)icariin can promote the osteogenic differentiation of TDSCs in rats,which is correlated with the concentration of icariin,and 10^-5mol/l is the best.(2)Icariin can up regulate the expression of Runx2 and OSX m RNA,which may be one of the mechanisms of icariin promoting osteogenic differentiation.(3)Icariin can be up-regulated?-The expression of catenin protein was positively correlated with icariin concentration and correlated with Wnt/?-Catenin pathway may play a leading role in osteogenic differentiation.