权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

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

免疫细胞引起的骨吸收机制的分子遗传学分析

基本信息

批准号：
17592078
负责人：
FUKUNAGA Jyoji
金额：
$ 2.24万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17592078/
关键词：
osteoporosis Immunosuppresant agents T, B cell Bone metabolism marker T, B細胞

项目摘要

(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

(An实验方法）使用1）FK 506的动物实验模型的制作：对实验组和对照组用等渗氯化钠溶液腹膜内注射FK 506，每天4周，持续6周，其中实验组为1龄ICR雄性小鼠。2)形态学观察：取胫骨石蜡切片，免疫组化染色见一条线，观察B细胞。接下来，我比较了与之对应的TRAP染色图像，并观察了对照组和实验组的变化。此外，大肠组织产生组织移植物并进行组织学观察。3)B细胞与破骨细胞关系的培养实验：破骨细胞的形成聚集了一个成髓细胞和第一个成骨细胞样细胞，并添加了活性形式的维生素D。我从一龄ICR雄性小鼠的肠系膜淋巴结中挑选了一个七周，我将一个CD 45 R阳性细胞分离并收集了它。 ...更多信息加入B细胞与B细胞直接接触或与滤过器共存培养体系，检测是否有某些因素间接促进破骨细胞的形成，并以破骨细胞的形成来评价。4)我收集了比胫骨、股骨和我进行TRAP染色的成髓细胞，并在剂量后，观察了破骨细胞数量的测量，培养10天后的形式分化能力。5)取FK剂量组小鼠肠系膜淋巴结（MLN），用流式细胞仪（FACS）分析。（结果/考虑）对于·对照组的免疫组织化学染色图像和与之对应的TRAP染色图像，B细胞通常打开，并且未识别I积累的TRAP阳性的状态和破骨细胞。然而，对于实验组，在骨表面上存在TRAP阳性的破骨细胞，并且与对照组相比增加。当我试着把两者放在一起时，B细胞似乎聚集在TRAP阳性的破骨细胞附近。通过培养实验，比较接触组与非接触组TRAP阳性的多核样破骨细胞的形成数量，TRAP阳性的多核样破骨细胞明显增多。IL-6和TNF-α在对照组中的I值较低，而在单用FK 506的人群中的I值较高。换句话说，这表明炎性细胞因子比FK 506对照组增加。FK 506治疗组的CD 4 + CD 45 RBloCD 25+（Treg）较对照组显著降低。FK 50 G治疗组中，CD 4 + CD 45 RBhiCD 25-比对照组有增加的趋势。少