Protection Of Icariin And Tetracycline On The Impairment Of Rat Bone Mesenchymal Stem Cells Induced By Glucocorticoid In Vitro

ObjectiveTo establish the model of rat bone mesenchymal stem cells impairmentinduced by glucocorticosteroid; To investigate the effect of icariin andtetracycline on the impairment of rat bone mesenchymal stem cells byglucocorticosteroid.MethodsRat BMSCs were isolated cultured in vitro. The model of BMSCsimpairment induced by glucocorticosteroid was established. There were fivegroups in vitro experiment, the first was normal control group, the second groupwas dexamethasone control group (dexamethasone10-8mol/L), the third groupwas the ICA group (ICA20ng/ml+dexamethasone10-8mol/L), the fourth groupwas the tetracycline group (tetracycline100μg/ml+dexamethasone10-8mol/L),the last group was the combined group (ICA20ng/ml+tetracycline100μg/ml+dexamethasone10-8mol/L), The proliferation of BMSCs wasevaluated by MTT assay at the different time points. The cells growth curve wasdraw by the results. The apoptosis of each group were measured by acridineorange/ethidium bromide fluorescence staining. The Bcl-2and Bax weremeasured by immunohistochemical method. The Caspase-3were measured byWestern blotting.ResultsDexamethasone inhibited cell proliferation. With the increase of dose and time dependent, the effect was enhanced. The Dexamethasone can induceapoptosis.MTT results OD values of the combined group compared with other groups,the combination protective effect of stronger than others, the gap is statisticallysignificant(p<0.05).Acridine orange/ethidium bromide fluorescence staining results of thecombined group apoptosis decreased, the gap is statistically significant(p<0.05).The immunohistochemical results of the combined group with other Bcl-2increased expression of Bax expression decreased, the gap is statisticallysignificant(p <0.05).Western blotting the results of the combined group Caspase-3expressioncompared with other groups decreased expression of the gap was statisticallysignificant(p <0.05).ConclusionsIcariin and Tetracycline can protect the impairment of BMSCs induced byglucocorticosteroid. our study provided some experimental pharmaco-foundation for intervention therapy of early osteonecrosis.