The Study Of The Mechanism On The Influence Of Hepcidin And Iron Ion Out Of Osteoblast On Calcium Transportation

The abnormality of bone metabolism is the cause of osteoporosis. Recently,the study of the relationship between ferric and bone metabolism is enheartening and joyful. Hepcidin is a type of antimicrobial polypeptide which is rich in aminothiopropionic acid.In 2001, Park separated this polypeptide from urina hominis, and named it Hepcidin.Many researches show that Hepcidin can regulate ferric metabolism as one type of hormonelike substance.Our purpose is to study the influence of Hepcidin and iron ion concentrations change out of hFOB1.19 cells on Ca2+ transportation. We use iron ion and calcium ion as vehicles to study the the relationship among Hepcidin ,ferric metabolism and bone metabolism at cellular level.The result of this study could be the base of the instructions for curing osteoporosis by Hepcidin. The hFOB 1.19 was cultured in the nutrient medium with iron ion and Hepcidin in different concentrations, then the change of calcium ion concentrations inside hFOB 1.19 was tested in different experimental session to study the influence of Hepcidin and iron ion out of osteoblast on calcium transportation.This study is divided into three parts as below:Part one: The mechanism on the influence of iron ion out of osteoblast on calcium transportationObjective To study the influence of iron ion concentrations change out of hFOB1.19 cells on calcium transportation. Methods The hFOB 1.19 was cultured at 34℃for 3-4 days, being digested with trypsin, then placed in 12 hole the cultivate board. Add D2H2O into the control group and DFO or FAC of different concentration into the experiment group,then confocal laser scanning microscope (CLSM) was used 2h later.Results Diversion of Ca2+ into cells was increased with the decrease of Fe3+ concentration out of hFOB 1.19 cells(the final consistence of DFO was 200-300 umol/L),but was hindered with the increase of Fe3+ concentration out of them(the final consistence of FAC was 50-100 umol/L) as observed under CLSM. Conclusion The Ca2+ transportation was increased with the decrease of Fe3+ concentration out of hFOB 1.19 cells but was hindered with the increase of Fe3+ concentration out of them.Part Two: The effect of time factors on the influence of iron ion out of osteoblast on calcium transportation 0bjective To study whether there is time dependability on the influence of iron ion concentrations change out of hFOB1.19 cells on calcium transportation. Methods The hFOB 1.19 was cultured at 34℃for 3-4 days, being digested with trypsin, then placed in 12 hole the cultivate board. Add D2H2O into the control group and DFO or FAC of different concentration into the experiment group,then confocal laser scanning microscope (CLSM) was used 2h and 48h later.Results Diversion of Ca2+ into cells was increased with the decrease of Fe3+ concentration out of hFOB 1.19 cells(the final consistence of DFO was 200-300 umol/L) and as the interfering time going by, the diversion of more Ca2+ into cells was increased . However, Diversion of Ca2+ into cells was hindered with the increase of Fe3+ concentration out of them(the final consistence of FAC was 50-100 umol/L) and as the interfering time going by, the diversion of more Ca2+ into cells was hindered as observed under CLSM. Conclusion The Ca2+ transportation was increased with the decrease of Fe3+ concentration out of hFOB 1.19 cells but was hindered with the increase of Fe3+ concentration out of them. There is time dependability on the influence of iron ion concentrations change out of hFOB1.19 cells on calcium transportation. Part Three: A preliminary study of the mechanism on the influence of hepcidin out of osteoblast on calcium transportationObjective To study the influence of hepcidin concentrations change out of hFOB1.19 cells on calcium transportation. Methods The hFOB 1.19 was cultured at 34℃for 3-4 days, being digested with trypsin, then placed in the culture board. Add D2H2O into the control group and Hepcidin of different concentration into the experiment group:H1 group( final concentration is 10nM/L) and H2 group(final concentration is 50nM/L). Confocal laser scanning microscope (CLSM) was used in this study 24 hours later .Results The fluorescence intensity of calcium ion in hFOB 1.19 from control group is 56.38±36.47,however, that is 68.01±32.90 and 154.06±55.62 of H1 and H2 group ,respectively(P<0.05). Conclusion The Ca2+ transportation was increased with the increase of Hepcidin concentration out of hFOB 1.19 cells and the tendency of diversion of Ca2+ into cells was extremely significant.