Effects of IGF-IR iRNA on the growth of condylar cartilage by electric pulse delivery

IGF-IR iRNA 对电脉冲递送髁软骨生长的影响

• 批准号: 17592131
• 负责人: SUZUKI Shoichi
• 金额: $ 2.24万
• 依托单位: Tokyo Medical and Dental 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-17592131/
• 关键词: Mandibular condyle; IGF-I; apoptosis; IGFBP3; caspase 3; アンチセンス; IGF-IR; 下顎頭軟骨細胞

The purpose of this study was to examine the role of endogenous insulin-like growth factor I (IGF-I) in promoting survival of cells of the mandibular condyle in culture. Mandibular condyles dissected from 12-day-old rats were cultured for 1, 3 or 5 days in medium containing antisense oligodeoxynucleotide (AS-ODN) or sense-ODN (S-ODN) for IGF-I. Quantitative real-time RT-PCR analysis showed that the levels of IGF-I and IGF binding protein (IGFBP)3 mRNAs in tissues treated with AS-ODN were significantly decreased compared with control and S-ODN group. On days 3 and 5, the number of denatured cells was observed in the undifferentiated mesenchymal cell layer. These cells were TUNEL positive and confirmed to be apoptotic by electron microscopic observation. Immunoblotting revealed that expression of cleaved caspase3 was increased with AS-ODN. These results indicate that the cells in the undifferentiated mesenchymal cell layer of mandibular condyle require IGF-I for survival.

本研究旨在探讨内源性胰岛素样生长因子I（IGF-I）在促进培养的下颌骨髁突细胞存活中的作用。取12日龄大鼠下颌骨髁状突，分别在含有IGF-Ⅰ反义寡核苷酸（AS-ODN）或正义寡核苷酸（S-ODN）的培养液中培养1、3或5d。实时荧光定量RT-PCR结果显示，与对照组和S-ODN组相比，AS-ODN组IGF-I和IGFBP 3 mRNA水平显著降低。在第3天和第5天，在未分化间充质细胞层中观察到变性细胞的数目。电镜观察证实细胞凋亡，TUNEL阳性。免疫印迹结果显示，AS-ODN可增加caspase 3的表达。提示髁突未分化间充质细胞层的细胞需要IGF-I才能生存。