Interleukin-1 (IL-1) and insulin-like growth factor-I (IGF-I), which have opposing effects on matrix metabolism within articular cartilage, are thought to play prominent roles in the pathogenesis of osteoarthritis. To better understand the link between these anabolic (IGF-I) and catabolic (IL-1) stimuli, we examined exogenous IL-1 regulation of the IGF-I signaling system of articular chondrocytes (ACs). Equine ACs from non-arthritic stifle joints were expanded in monolayer culture, encapsulated for 10 days in alginate beads, and stimulated as high-density monolayers with recombinant equine IL-1beta (0, 1, 10 ng/ml) for 48 h. IL-1beta enhanced expression of IGF-IR levels, as determined by both [125I]-IGF-I binding studies and Western blotting, while reducing the concentration of endogenous IGF-I detected in conditioned media by radioimmunoassay. Western ligand blotting revealed that chondrocytes primarily secreted IGF binding proteins (IGFBPs) with molecular weights of 28-30 and 32-34 kDa, which were identified as IGFBPs 5 and 2, respectively, and that IL-1beta treatment diminished IGFBP-2, the prominent homolog in conditioned media. Northern blot analysis suggested IL-1beta regulation of IGF-I and, to some extent, IGF-IR was mediated by transcription; however, the cytokine did not affect IGFBP-2 expression. To test for evidence of proteolysis by matrix metalloproteinases (MMPs), additional cultures were co-incubated with inhibitors for MMPs 2/9, 3, and 8. IGFBP-2 suppression was partially reversed by gelatinase (MMP-2/9) inhibition. In summary, these findings further delineate the role of IL-1 as a key regulator of the IGF-I system within articular cartilage, demonstrating that regulation occurs through both direct (transcriptional) and indirect (proteolytic) mechanisms.

译文

Interleukin-1 (IL-1) 和胰岛素样生长因子-I (igf-i) 对关节软骨内基质代谢具有相反作用,被认为在骨关节炎的发病机理中起着重要作用。为了更好地了解这些合成代谢 (igf-i) 和分解代谢 (IL-1) 刺激之间的联系,我们检查了关节软骨细胞 (ACs) 的igf-i信号系统的外源性IL-1调节。来自非关节炎窒息关节的马ACs在单层培养中扩增,在藻酸盐珠中封装10天,并用重组马IL-1beta (0,1,10 ng/ml) 刺激为高密度单层48小时。通过 [125I]-igf-i结合研究和Western印迹测定,IL-1beta增强了igf-ir水平的表达,同时降低了通过放射免疫测定法在条件培养基中检测到的内源性igf-i的浓度。Western配体印迹显示,软骨细胞主要分泌分子量为28-30和32-34 kDa的IGF结合蛋白 (IGFBPs),分别被鉴定为IGFBPs 5和2,并且IL-1beta治疗减少了IGFBP-2,这是条件培养基中的主要同源物。Northern印迹分析表明igf-i的IL-1beta调节,并且在某种程度上,igf-ir是通过转录介导的; 然而,细胞因子不影响IGFBP-2的表达。为了测试基质金属蛋白酶 (mmp) 蛋白水解的证据,将另外的培养物与mmp 2/9、3和8的抑制剂共同孵育。IGFBP-2抑制被明胶酶 (MMP-2/9) 抑制部分逆转。总之,这些发现进一步描述了IL-1作为关节软骨内igf-i系统的关键调节因子的作用,表明调节通过直接 (转录) 和间接 (蛋白水解) 机制发生。

+1
+2
100研值 100研值 ¥99课程
检索文献一次
下载文献一次

去下载>

成功解锁2个技能,为你点赞

《SCI写作十大必备语法》
解决你的SCI语法难题!

技能熟练度+1

视频课《玩转文献检索》
让你成为检索达人!

恭喜完成新手挑战

手机微信扫一扫,添加好友领取

免费领《Endnote文献管理工具+教程》

微信扫码, 免费领取

手机登录

获取验证码
登录