Analysis of molecular genetics to the bone resorption mechanism that an immune cell causes

(An Experiment Method)Manufacture of the animal experiment model who used1)FK506：I did intraperitoneal injection of FK506 to an experimental group with isotonic sodium chloride solution in control group with an ICR maleness mouse of an instar day after day for four weeks for six weeks。2)form observation：There was a line by immunohistochemistry dyeing I made a paraffin graft of a tibia，and to observe B cell。I compared a TRAP dyeing image corresponding to it next and observed control group and a change in an experimental group each.Furthermore，a large intestine organization produced an organization graft and performed histologic observation。3)A culture experiment to examine relations of B cell and osteoclast：The formation of osteoclast gathered a myeloblast and first osteogenetic cell-like cell and added active form vitamin D.I picked out a mesentery lymph node of an ICR maleness mouse of an instar for seven weeks and I divided a CD45R positive cell and collected it。I……More added B cell and B cell came in contact directly or and I used a filter for coexistence culture system to examine whether some factors promoted the formation of osteoclast indirectly and evaluated it by the formation of osteoclast.4)I gathered a myeloblast than a tibia，the thighbone and I performed TRAP dyeing and，after the dosage，observed a measurement of the number of the osteoclast，form differentiation ability after culture of ten days。5)I picked out mesentery lymph node(MLN)of an FK dosage mouse and analyzed it in FACS.6)I measured serum cytokine in Bio-Plex.(A result/consideration)With an immunohistochemistry dyeing image of·control group and a TRAP dyeing image corresponding to it，B cell opened generally，and a state and osteoclast of TRAP positive which I accumulwere not ognized。However，with an experimental group，there was osteoclast of TRAP positive on the surface of a bone and increased in comparison with control group。The B cell seemed to accumulate it near osteoclast of TRAP positive when I tried to put both together.By a·culture experiment，I compared contact group with non-contact group with the number of the formation of an osteoclast of many nucleuses-like cell with TRAP positive，and osteoclast cells of many nucleuses increased with TRAP positive significantly。As well as·interleukin-6 and TNF-α，I showed a low value in control group and showed a high price in the crowd who administered only FK506。In other words it was suggested that inflammatory cytokine increased than control by FK506。·CD4+CD45RBloCD25+(Treg)decreased in a FK506 treated group more significantly than control group.A tendency to increase had CD4+CD45RBhiCD25-than control group in a FK50G treated group.Less：Less

(An experiment method) Manufacture of the animal experiment model who used 1) FK506 : I did intraperitoneal injection of FK506 to an experimental group with isotonic sodium chloride solution in control group with an ICR maleness mouse of an instar day after day for four weeks for six weeks. 2) form observation : There was a line by immunohistochemistry dyeing I made a paraffin graft of a tibia, and to observe B cell. I compared a TRAP dyeing image corresponding to it next and observed control group and a change in an experimental group each. Furthermore, a large intestine organization produced an organization graft and performed histologic observation. 3) A culture experiment to examine relations of B cell and osteoclast : The formation of osteoclast gathered a myeloblast and first osteogenetic cell-like cell and added active form vitamin D. I picked out a mesentery lymph node of an ICR maleness mouse of an instar for seven weeks and I divided a CD45R positive cell and collected it. I … More added B cell and B cell came in contact directly or and I used a filter for coexistence culture system to examine whether some factors promoted the formation of osteoclast indirectly and evaluated it by the formation of osteoclast. 4) I gathered a myeloblast than a tibia, the thighbone and I performed TRAP dyeing and, after the dosage, observed a measurement of the number of the osteoclast, form differentiation ability after culture of ten days. 5) I picked out mesentery lymph node (MLN) of an FK dosage mouse and analyzed it in FACS.6) I measured serum cytokine in Bio-Plex.(A result / consideration) With an immunohistochemistry dyeing image of・control group and a TRAP dyeing image corresponding to it, B cell opened generally, and a state and osteoclast of TRAP positive which I accumulated were not recognized. However, with an experimental group, there was osteoclast of TRAP positive on the surface of a bone and increased in comparison with control group. The B cell seemed to accumulate it near osteoclast of TRAP positive when I tried to put both together. By a・culture experiment, I compared contact group with non-contact group with the number of the formation of an osteoclast of many nucleuses-like cell with TRAP positive, and osteoclast cells of many nucleuses increased with TRAP positive significantly. As well as・interleukin-6 and TNF-α, I showed a low value in control group and showed a high price in the crowd who administered only FK506. In other words it was suggested that inflammatory cytokine increased than control by FK506.・CD4+CD45RBloCD25+ (Treg) decreased in a FK506 treated group more significantly than control group. A tendency to increase had CD4+CD45RBhiCD25-than control group in a FK50G treated group. Less