Effects Of Hepcidine On The Proliferation And Expressions Of Genes In MC3T3-E1 Osteoblasts Of Mice

Along with accelerated "aging" process o,the incidence of osteoporosis is rising. Osteoporosis is a bone metabolic disorder chacterized by damage of bone tissue microstructure, decrease of bone mineral composition and the proportion of bone matrix, bone thinning, reduction in the number of trabecular bone, bone fragility growing and increased fracture risk. Research about interaction of the iron metabolism and bone metabolism was found in the early 21st century. The evidents are based on twoaspect. The first is that abnormal iron metabolism in various was diseases associated with abnormal bone metabolism. And the other one is osteoporosis associated with iron metabolism.Research on interaction of iron metabolism and bone metabolism attracted more and more attention currently. Hepcidin, a key regulator of iron metabolism, is a kind of small molecule peptide hormones synthesized by the liver. Hepcidin is the main regulator of Iron homeostasis, which is mainly synthesized in the liver cells and secreted into blood. Hepcidin transmits the signal to small intestine, regulating the iron absorption of small intestinal.Hepcidin was used to interventi osteoblasts in our research. Cell proliferation and change of osteoblast osteocalcin,and osteoprotegerin genes were observed to clarify the effect of steady-state of iron on bone metabolic disorder such as Osteoporosis. This study is divided into three parts as below:Part one: Effects of hepcidine on the proliferation in MC3T3-E1 osteoblasts of miceobjective To investigate the effects of hepcidine on the proliferation in MC3T3-E1 osteoblasts of mice in different concentrations of hepcidin。Methods After MC3T3-E1 mice cells were cultured with different concentrations of hepcidine,FAC,DFO for 24 hours,48 hours,72 hours,we evaluated the proliferation of cell using MTT. Results After the intervention of different concentrations of hepcidine,DFO,FAC,The result of MTT showed that the growth were upgraded by the hepcidine,DFO concertrations adding, the growth were downgraded by the FAC concertrations adding, which showed a statistical difference of significance (P <0.05).Conclusion Excess iron inhibits cell growth,and Hepcidine,DFO upgraded cell growth,with a dose-dependent effect.Part two: Effects of hepcidine on expressions of osteoprotegerin and osteocalcin genes in MC3T3-E1 osteoblasts of miceobjective To investigate the effects of hepcidine on osteoprotegerin (OPG)and osteocalcin(BGP)gene expressions in MC3T3-E1 osteoblasts of mice。Methods After hepcidine of different concentrations being added in MC3T3-E1 Mice cells cultures in vitro (100nmol/ml, 200nmol/ml, 300nmol/ml) for 72 hours ,we evaluated the expression of OPG, BGP mRNA using RT-PCR. Results The result of RT-PCR showed that the BGP mRNA and OPG mRNA expression all occurred after the intervention of hepcidine of different concentrations;And the density ratios of OPG mRNA and BGP mRNA shown on RT-PCR differed according to the concentrations ,which showed a statistical difference of significance (P <0.05).Conclusion Hepcidine upgraded the expression of BGP and OPG mRNA in mice MC3T3-E1 cells ,with a dose-dependent effect.Part three: Effects of desferrioxamine on the function of MC3T3-E1 osteoblasts of miceobjective To investigate the effects of hepcidine on osteoprotegerin (OPG) and osteocalcin(BGP)gene expressions in MC3T3-E1 osteoblasts of mice。Methods After different concentrations of desferrioxamine being added in MC3T3-E1 mice cells cultures in vitro (50μmol/L,100μmol/L) for 48 hours ,we evaluated the expression of OPG, BGP mRNA in MC3T3-E1 mice cells using RT-PCR. Results The result of RT-PCR showed that the BGP mRNA and OPG mRNA expression all occurred after the intervention of desferrioxamine of different concentrations;And the density ratios of OPG mRNA and BGP mRNA shown on RT-PCR differed according to the concentrations ,which showed a statistical difference of significance (P <0.05).Conclusion Desferrioxamine upgraded the expression of BGP and OPG mRNA in mice MC3T3-E1 cells ,with a dose-dependent effect.