development of novel conservative therapies for joint disease under controlled HSP expression-usage of HSP-inducing agents and thermotherapy-

开发针对受控 HSP 表达的关节疾病的新型保守疗法 - 使用 HSP 诱导剂和热疗 -

• 批准号: 17390419
• 负责人: KUBO Toshikazu
• 金额: $ 9.98万
• 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 Scientific Research (B)
• 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/grant/KAKENHI-PROJECT-17390419/
• 关键词: cartilage; cartilage metabolism; thermotherapy; microwave; heat shock protein; osteoarthritis; 関節軟骨; 軟骨細胞; HSP70; グルタミン; アポトーシス; 熱ストレス; SNP; 軟骨保護作用

Heat stimulation was applied to the knee joints of Japanese white rabbits for 20 min using a microwave (MW) applicator (2.45-GHz, 0-80 W). After 8-72 hrs, the articular cartilage was removed from the knee joints and proteins and total RNA were extracted. As controls, the knee joints without heat stimulation were analyzed. Expression of heat shock protein (HSP) 70 was confirmed by real-time PCR and Western blotting. Expression of proteoglycan core protein (PG) and type II collagen (Col II) were quantified using real-time PCR to assess cartilage matrix metabolism. Compared to controls, HSP70 expression was higher with more than 40 W of heat stimulation. The expression of PG and Col II mRNA were higher with more than 20 W of heat stimulation and peaked with 40 W. The highest expression of HSP70, PG, and Col II mRNA were observed 8hrs, 48hrs, and 72hrs after heat stimulation, respectively. On the other hand, the highest HSP70 protein expression was observed 24 h after heat stimulation. When quercetin was used to inhibit the induction of HSP70 expression, PG mRNA expression did not increase. The present study demonstrated that appropriate heat stimulation increased the expressions of extracellular matrix gene, which might be partially mediated by HSP70. These findings showed that controlling HSP70 expression in the cartilage using thermotherapy might offer a novel strategy for suppressing cartilage degeneration in osteoarthritis.

使用微波（MW）施加器（2.45-GHz，0-80 W）对日本白色家兔的膝关节施加热刺激20 min。8-72小时后，从膝关节取出关节软骨，提取蛋白质和总RNA。作为对照，分析无热刺激的膝关节。实时荧光定量PCR和Western blotting检测热休克蛋白70的表达。蛋白多糖核心蛋白（PG）和II型胶原（Col II）的表达采用实时PCR定量评估软骨基质代谢。与对照组相比，HSP 70的表达高于40 W的热刺激。PG和Col Ⅱ mRNA表达在热刺激20 W以上时升高，40 W时达到高峰。HSP 70、PG和Col Ⅱ mRNA的表达分别在热刺激后8h、48 h和72 h达到高峰。另一方面，观察到热刺激后24 h的最高HSP 70蛋白表达。槲皮素抑制HSP 70的诱导表达后，PG mRNA的表达没有增加。本研究表明，适当的热刺激可促进细胞外基质基因的表达，其作用可能部分由热休克蛋白70介导。这些发现表明，使用热疗控制软骨中HSP 70的表达可能为抑制骨关节炎中的软骨退变提供了一种新的策略。

项目成果

Effects of heat stimulation via microwave applicator on cartilage matrix gene and HSP70 expression in the rabbit knee joint

• DOI: 10.1002/jor.20421
• 发表时间: 2008-01
• 期刊: Journal of Orthopaedic Research
• 影响因子: 2.8
• 作者: H. Tonomura;Kenji A. Takahashi;O. Mazda;Y. Arai;M. Shin-Ya;A. Inoue;K. Honjo;T. Hojo;J. Imanishi;T. Kubo