Effects of naringenin on apoptosis of osteoblasts mediated by oxidative stress and its action mechanism

doi:10.3969/j.issn.1002-7386.2016.19.008

Abstract：Objective To observe the effects of naringenin on apoptosis of osteoblasts mediated by oxidative stress, and to explore its action mechanism.Methods The osteoblasts were separated and cultured in vitro.The experiment was divided into 5 groups:blank control group,simple H2O2 group, H2O2 +naringenin low-does group (0.1μmol/L), H2O2 +naringenin medium-dose group ( 1μmol/L ) and H2 O2 +naringenin high-dose group ( 10μmol/L ) .The cell viabilities of osteoblasts were detected by MTT assay.The cell apoptosis rates were detected by flow cytometry.The levels of reactive oxygen species (ROS) were observed by fluorescence microscopy.MDA contents were measured by colorimetric technique.The expression levels of Bcl-2, Bax and Caspase-3 mRNA and protein were detected by Real time-PCR and Western Blot, respectively.Results As compared with those in blank control group,the cell viabilities in simple H2 O2 group were obviously decreased,however, the cell apoptosis rates and the levels of ROS and MDA were significantly increased, meanwhile, the expression levels of Bcl-2 mRNA and protein were obviously down-regulated, however, the expression levels of Bax and Caspase-3 mRNA and protein were significantly up-regulated ( P <0.05).As compared with those in simple H2O2 group, the cell viabilities in naringenin groups pretreated for 24 hours were obviously increased, and the cell apoptosis rates and the levels of ROS and MDA were significantly decreased, moreover, the expression levels of Bcl-2 mRNA and protein were obviously up-regulated, however, the expression levels of Bax and Caspase-3 mRNA and protein were significantly down-regulated,with a dose-dependent manner ( P <0.05).Conclusion Naringenin can promote proliferation of osteoblasts and inhibit apoptosis of osteoblasts mediated by oxidative stress, and its action mechanism may be correlated to up-regulating the expressions of Bcl-2 and down-regulating the expressions of Bax and Caspase-3.