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Expression and role of stress protein at the injury of joint cartilage

应激蛋白在关节软骨损伤中的表达及作用

基本信息

批准号：
05454411
负责人：
KUBO Toshikazu
金额：
$ 1.54万
依托单位：
Kyoto Prefectural University of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05454411/
关键词：
Osteoarthritis Heat shock protein Hydrostatic pressure PCR Cytokine Black mouse 免疫組織化学 in situ hybridization

项目摘要

1. Localization of heat shock protein in osteoarthritic cartilage : by immunohistochemistry, in normal and mild osteoarthritic cartilage, HSP70 was only localized in the chondrocytes of the superficial layr where safranin O staining was reduced. In moderate and severe osteoarthritic cartilage, the chondrocytes with positive staining were recognized to deeper layrs. The distribution of positively stained chondrocytes was correlated to the severity of osteoarthritis (OA).2. Influence of hydrostatic pressure (HP) on chondrocytes : Chondrocyte like cells (HCS2/8) were subjected to HP treatment ranging from 0Mpa to 50Mpa. Expression of bFGF and IGF-I mRNA was enhanced by HP as shown by PCR analysis. Expression of TGF beta1 mRNA was enhanced after 5Mpa HP and reduced after the 50 Mpa HP.Expression of HSP70 mRNA was markedly enhanced after 50 Mpa HP.Incorporation of 35S sulfate after 5 Mpa HP treatment increased significantly compared with the control. However, it was reduced after 50 Mpa HP treatment.3. Analysis of spontaneous osteoarthritic mouse by PCR : Six-month-old C57 black mouse (Silberberg) which is developing spontaneous OA were used. Total RNA was extracted from the proximal tibial cartilage and PCR for various cytokines and HSPs was performed. Statistical analysis indicated that IL-6, IFN-gamma and HSPs (47,70,86) were strongly expressed in comparison with control mouse.

1.热休克蛋白在骨关节炎软骨中的定位：免疫组化结果显示，在正常和轻度骨关节炎软骨中，HSP 70仅定位于浅层软骨细胞，番红O染色减弱。在中、重度骨关节炎软骨中，染色阳性的软骨细胞向更深的层识别。阳性染色软骨细胞的分布与骨关节炎（OA）的严重程度相关.静水压对软骨细胞的影响：软骨细胞样细胞（HCS 2/8）在0 ~ 50 Mpa的压力下培养。PCR分析显示HP可促进bFGF和IGF-I mRNA的表达。5 Mpa高压处理后，TGF β 1 mRNA表达增强，50 Mpa高压处理后，TGF β 1 mRNA表达减弱，HSP 70 mRNA表达增强，35 S硫酸盐掺入量增加。而50 MPa高压处理后，则有所降低.通过PCR分析自发性骨关节炎小鼠：使用正在发展自发性OA的6月龄C57黑小鼠（西尔伯贝格）。从胫骨近端软骨中提取总RNA，并进行各种细胞因子和热休克蛋白的PCR。统计学分析表明，与对照小鼠相比，IL-6、IFN-γ和HSPs（47、70、86）强烈表达。