TREATMENT OF ARTHRITIS WITH ANTISENSE ORIGONUCLEOTIDE FOR TRANSCRIPTION FACTOR NF-κB

用转录因子 NF-κB 反义原核苷酸治疗关节炎

• 批准号: 11671446
• 负责人: TAKAGI Toshitaka
• 金额: $ 2.24万
• 依托单位: YOKOHAMA CITY UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671446/
• 关键词: NF-κB; antisense oligonucleotide; mouse collagen-induced arthritis; AP-1; rheumatoid arthritis; mouse air-pouch; apoptosis; 免疫組織化学; 軟骨細胞

1. DBA/1J male mice were immunized with bovine type II collagen emulsified with Freund's incomplete adjuvant as a model for rheumatoid arthritis. Five mice were sacrificed at 3,5,7 weeks after the immunization. Knee joints were fixed in 10% neutralized formic acid, decalcified with 0.5% EDTA and embedded in paraffin. Histological changes were evaluated in sections stained with hematoxylin-eosin and immunostaining with rabbit anti-NF-κB p65 polyclonal antibodies (Santa Cruz) using avidin-biotin complex method. NF-κB was positive in synovial lining cells and chondrocytes. The mean rates of NF-κB-positive chondrocye at 3,5 and 7 weeks after the immunization were 43.7%, 68.0% and 79.5%. The rate of mice without treatment was 51.1%.2. Antisense oligonucleotide for NF-κB designed as 5'-GAA ACAGATCGTCCATGGT or a mismatched sense designed as 5'-GAAACAGATCGTCTATGGT was intraperitonealy injected into the DBA/1J mice 4 weeks after the immunization with type II collagen for 2 weeks every day. Significant inhibition of swelling and redness of pows was not observed. These findings indicated two critical problems as follows. One was a delivery system of the oligonucleotide, and the other was an effect of another transcription factor AP-1. After then we observed AP-1 expression in the lining cells of mouse air-pouch model induced by lipopolysaccharide. Further investigation on these points will be required.

1.弗氏不完全佐剂乳化牛II型胶原蛋白免疫DBA/1J雄性小鼠，作为类风湿性关节炎模型。免疫后3、5、7周处死5只小鼠。将膝关节固定在10％中和的甲酸中，用0.5％EDTA脱钙并包埋在石蜡中。使用抗生物素蛋白-生物素复合物方法，用苏木精-伊红染色和用兔抗NF-κB p65多克隆抗体(Santa Cruz)免疫染色的切片评估组织学变化。滑膜衬里细胞和软骨细胞中 NF-κB 呈阳性。免疫后3、5、7周NF-κB软骨细胞阳性率平均分别为43.7%、68.0%和79.5%。小鼠未治疗率为51.1%。2。在II型胶原免疫后4周，每天将设计为5'-GAA ACAGATCGTCCATGGT的NF-κB反义寡核苷酸设计为5'-GAA ACAGATCGTCCATGGT或设计为5'-GAAACAGATCGTCTATGGT的错配正义寡核苷酸设计为DBA/1J小鼠腹膜内注射，连续2周。没有观察到对战俘肿胀和发红的显着抑制。这些发现表明了以下两个关键问题。一个是寡核苷酸的递送系统，另一个是另一种转录因子AP-1的作用。然后我们观察脂多糖诱导的小鼠气囊模型内衬细胞中AP-1的表达。需要对这些点进行进一步调查。

项目成果

Takagi T, Koshino T, Hayashi T, Sakurai S, Eguchi J: "Roles of NF-κB in destruction of articular cartilage. (in Japanese)."THE BONE. 14. 73-75 (2000)

Takagi T、Koshino T、Hayashi T、Sakurai S、Eguchi J：“NF-κB 在关节软骨破坏中的作用。（日语）。”THE BONE。

桜井真一: "Lipopolysaccharide反復投与によるマウスair-pouch表層細胞のapoptosisと浸出液中soluble Fasの出現"日本整形外科学会誌. 73. S1645 (1999)

Shinichi Sakurai：“由于重复施用脂多糖，导致小鼠气囊表面细胞凋亡和渗出液中出现可溶性 Fas”，日本骨科学会杂志 73. S1645 (1999)。

Sakurai S, Takagi T, Okamoto R, Hayashi T, Matsukura Y, Sato MI, Sato M, Kurosaka N, Eguchi J, Ogura K, Koshino T: "Apoptosis of the lining cells and up-regulation of soluble Fas in the exudates of the mouse air pouch induced by lipopolysaccharide. (in Ja

Sakurai S、Takagi T、Okamoto R、Hayashi T、Matsukura Y、Sato MI、Sato M、Kurosaka N、Eguchi J、Ogura K、Koshino T：“衬里细胞的凋亡和渗出物中可溶性 Fas 的上调

高木敏貴: "関節軟骨破壊とNF-κB"THE BONE. 14・3. 73-75 (2000)

高木俊树：“关节软骨破坏和 NF-κB”THE BONE 73-75 (2000)。

高木敏貴: "関節軟骨破壊とNF-κB"THE BONE. 14. 73-75 (2000)

Toshiki Takagi：“关节软骨破坏和 NF-κB”THE BONE 14. 73-75 (2000)。