Protective Effect Of Genistein On Hypoxic Injuries Of Osteoblasts And RBMSCs In Vitro

Genistein inhibits bone resorption and increased bone mineral density. In this study,potential of anti-hypoxia protection and promoting differentiation and mineralization ofgenistein was investigated in rat osteoblasts (ROB) with hypoxia exposure, administered atdoses of10-6mol/L,10-5mol/L and10-4mol/L and rat bone marrow stromal cells (rBMSCs)with hypoxia exposure, administered at doses of10-8mol/L,10-7mol/L and10-6mol/L.After36or48hours of hypoxia, cell viability, intracellular reactive oxygen species(ROS) level, lactate dehydrogenase (LDH) release, alkaline phosphatase (ALP) activity andcalcium deposition were measured. Flow cytometer analyzes apoptosis and cell cycle.Expression of proliferating cell nuclear antigen (PCNA) was investigated byimmunohistochemistry. Mineralized bone nodules were revealed by alizarin red staining andvon Kossa staining. Collagen I were determined by immunohistochemistry and westernblotting. Relative level of mRNA expression of hypoxia inducible factor1α (HIF-1α),anti-apoptotic gene Bcl-2, apoptotic gene Caspase-3and osteogenic growth factor Runx-2,BMP-2and Osterix (OSX) were quantitatively determined by RT-PCR.The results showed that genistein enhances cell viability and ALP activity, and decreasesROS level, LDH release and apoptosis, and expression of PCNA was inhibited, and cell cyclearrest in S phase. Genistein boosts HIF-1α, Bcl-2, Runx-2, BMP-2and Osterix (OSX) mRNAexpression, and inhibits Caspase-3mRNA expression. In addition, genistein improve area ofmineralized bone nodules and expression of collagen I in ROB and rMBSCs with hypoxia.In conclusion, genistein has potential of anti-hypoxia protection and enhance osteogenicdifferentiation of ROB and rBMSCs treated with hypoxia significantly.