Objective To investigate the effect of hypoxia inducible factor-1α(HIF-1α)on the osteogenic differentia-tion of mesenchymal stem cells(MSCs)C3H10T1/2 induced by bone morphogenetic protein 9(BMP9).Methods MSCs C3H10T1/2 cells line of the rats were cultured in vitro and divided into 5 groups:NC group(normal cultured C3H10T1/2 cells),Ad-GFP group(transfected with adenovirus control),Ad-BMP9 group(transfected with adenovirus over expression of BMP9),pcDNA3.1-mRFP group(transfected with Ad-BMP9 + pcDNA3.1 empty plasmid),pcD-NA3.1-HIF-1α group(transfected with Ad-BMP9 + HIF-1α over expression plasmid).Alkaline phosphatase(ALP)activity in C3H10T1/2 cells of each group was detected by ALP staining and activity detection kit;the calcium deposi-tion of C3H10T1/2 cells was detected by Alizarin Red S staining,and the absorbance(A)value was used for quanti-tative analysis;and the expressions of BMP9,HIF-1α,osteogenic differentiation markers osteocalcin(OC),osteopontin(OPN)and Runt related transcription factor2(Runx2)in C3H10T1/2 cells were detected by Western blot method.Results Compared with NC group and Ad-GFP group,the ALP activity,calcium deposition,A value and protein ex-pressions of BMP9,HIF-1α,OC,OPN and Runx2 in Ad-BMP9 group were increased(P<0.05).Compared with Ad-BMP9 group and pcDNA3.1-mRFP group,the ALP activity,calcium deposition,A value and protein expressions of BMP9,HIF-1α,OC,OPN and Runx2 in pcDNA3.1-HIF-1α group were increased(P<0.05).Conclusions HIF-1α can promote the osteogenic differentiation of MSCs C3H10T1/2 induced by BMP9,which may be related to the up-regulation of OC,OPN and their downstream Runx2 protein expression.
hypoxia inducible factor-1αbone morphogenetic protein 9mesenchymal stem cellsosteogenic differenti-ation
维普
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