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

Development of an osteoporosis model pro-culture by immunity restraint agent and molecular biology analysis of bone absorption mechanism

免疫抑制剂亲培养骨质疏松模型的建立及骨吸收机制的分子生物学分析

基本信息

批准号：
15592108
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：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15592108/
关键词：
osteoporosis Immunosuppresant agents osteoclast T, B細胞骨代謝マーカー FACS T細胞 3次元マイクロCT

项目摘要

Blood and urine analysis : Although the level of serum calcium remained mostly within the normal range for both groups, there was a significant increase in the level of urinary Ca for the FK506 treated group from Week 1 of administration. The level of blood osteocalcin for the FK506 treated group at Week 1 of administration was significantly higher than that for the control group, but there was no significant difference after that. Throughout the administration period, the sum of urinary PYD and Dpd for the FK506 treated group was significantly higher. Of the various bone resorption factors, the level of blood PTH for the FK506 treated group was significantly higher than that for the control group at Week 3 of administration and later.Cell culture : After acclimatization for a week, mice were randomly divided into two groups and were intraperitoneally injected every day either FK506 at a dose of 1mg/kg/day (FK506 treated group) or physiological saline at 10ml/kg/day (Control group). Pr … More ograf, a solution containing 5mg FK506/ml, was diluted in physiological saline to obtain a final concentration. bone marrow cells were collected by flushing femoral shafts using sterile needles. Cells plated in 24-well plates were cultured for 6 days in α-minimal essential medium supplemented with 10% fetal bovine serum, 100U/ml penicillin, and 100 μg/ml streptomycin, 30 ng/ml macrophage-colony stimulating factor, and 100 ng/ml soluble Receptor Activator of NFκB Ligand. TRAP assay : Staining for tartrate-resistant acid phosphatase(TRAP) activity, a leukocyte acid phosphatase assay kit was used. At the end of the culture period, TRAP positive cells exhibiting ≧3 nuclei were counted as mature osteoclasts and TRAP positive cells with <3 nuclei were considered osteoclast precursors. Resorption pit assay : Osteoclasts were seeded onto dentine slices placed in each well of 24-well plates, and after the cells were scraped off from the dentine slices, excavated pits were stained with acid hematoxylin Photographs were taken under a light microscope at ×40 magnification, and total areas of resorption pits were analyzed by the computer soft of NIH Image. Less

血液和尿液分析：尽管两组的血清钙水平大部分保持在正常范围内，但FK 506给药组的尿钙水平从给药第1周开始显著升高。给药第1周时，FK 506治疗组的血骨钙素水平显著高于对照组，但此后无显著差异。在整个给药期间，FK 506给药组的尿PYD和Dpd总和显著较高。在各种骨吸收因子中，在给药第3周及以后，FK 506治疗组的血PTH水平显著高于对照组。适应一周后，小鼠随机分为两组，每天腹腔注射FK 506 1 mg/kg（FK 506给药组）或生理盐水10 ml/kg/天（对照组）。PR ...更多信息用生理盐水稀释含5 mg FK 506/ml的奥曲肽溶液，以获得最终浓度。通过使用无菌针冲洗股骨干收集骨髓细胞。将接种在24孔板中的细胞在补充有10%胎牛血清、100 U/ml青霉素和100 μg/ml链霉素、30 ng/ml巨噬细胞集落刺激因子和100 ng/ml可溶性NFκB配体受体激活剂的α-最小必需培养基中培养6天。TRAP测定：抗酒石酸酸性磷酸酶（TRAP）活性染色，使用白细胞酸性磷酸酶测定试剂盒。在培养期结束时，显示<3个核的TRAP阳性细胞被计数为成熟破骨细胞，并且具有<3个核的TRAP阳性细胞被认为是破骨细胞前体。再吸收坑测定：将破骨细胞接种于24孔板各孔的牙本质切片上，刮除细胞后，用酸性苏木素染色，在放大40倍的光学显微镜下拍照，用NIH Image计算机软件分析牙本质吸收陷窝的总面积。少