摘要
目的观察人滑膜细胞Wnt/β-catenin信号通路激活后对其下游基因MMP-2、MMP-7和MMP-9的影响。方法采用GSK-3β选择性抑制剂SB-216763作用于正常人滑膜细胞,提取胞核蛋白β-catenin,采用Western blotting检测胞核蛋白β-catenin的表达变化;采用酶联免疫吸附法(ELISA)检测人滑膜细胞上清液中MMP-2、MMP-7和MMP-9的表达变化。结果 Western blotting结果显示,GSK-3β选择性抑制剂SB-216763作用于人滑膜细胞后,β-catenin在胞核蛋白中的表达水平明显升高,并随着GSK-3β选择性抑制剂作用时间的延长,β-catenin的表达逐渐增强,呈时间依赖性,而在其干预的第48小时,胞核蛋白β-catenin的表达变化最明显,是正常组滑膜细胞的5.8696倍,具有统计学意义(P<0.05);酶联免疫吸附法(ELISA)结果显示,与正常组滑膜细胞比较,SB-216763干预的滑膜细胞上清液中MMP-2、MMP-7和MMP-9的表达分别是正常滑膜细胞的4.6188、3.2443和2.9979倍,具有统计学意义(P<0.05)。结论①SB-216763成功激活人滑膜细胞Wnt/β-catenin信号通路;②MMP-2、7、9在人滑膜细胞Wnt/β-catenin信号通路激活后其表达显著升高,证明激活Wnt/β-catenin信号通路对MMP-2、MMP-7和MMP-9的表达有一定的调控作用。③Wnt/β-catenin信号通路在骨性关节炎患者中是激活的,而其对MMP-2、MMP-7和MMP-9的调控可能是导致关节软骨降解,促进骨关节炎形成的作用机制之一。该实验结果有助于从单一通路阐释Wnt/β-catenin信号通路对膝骨关节炎的作用机制。
[Objective] To observe the influence of activated human synovial cells Wnt B-catenin signaling pathway to its downstream gene MMP-2, MMP-7 and MMP-9. [Methods] The role of SB-216763, a selective inhibitor of GSK-3i3 in normal human synovial ceils, and then extract the nuclear proteins B- catenin by enzyme-linked immunosorbent assay using the Western Blot was used to detect nuclear proteins B-catenin expression changes; Detection of human synovial cell supernatant of MMP-2, MMP-7 and MMP-9 expression changes. [Results] The results showed that selective inhibitors of GSK-313 role of SB-216763 in human synovial cells, B-catenin in the nucleus protein expression levels ly increased, and as selectiveinhibitors of GSK-3B extension of time, B-catenin expression gradually increased in a time-dependent manner, enzyme-linked immunosorbent assay （ELISA） results show that compared with normal group, SB- 216763 intervention 48 h, nuclear proteins B-catenin expression is the most obvious change, it was 5.8696 with statistical significance （P 〈0.05）; compared with normal group ,the intervention of the synovial cell supernatant of MMP-2, MMP-7 and MMP-9 expression was respectively 4.6188, 3.2443, 2.9979, it was significantly higher than the normal synovial cell supernatant with statistical significance （P 〈0.05）. [ Conclusion] SB-216763 is successfully activated Wnt/I^-catenin signaling pathway in human synovial cells; MMP-2, MMP-7 and MMP-9 in human synovial cells Wnt/B-catenin signaling pathway activation and its expression was significantly elevated confirmed as the Wnt signaling pathway have a regulation on MMP-2, MMP-7 and MMP-9; the Wnt/B-catenin signaling pathway is activated in OA patients, the results of this experiment helps to explain the mechanism of Wnt/B-catenin signaling pathway knee osteoarthritis from a single pathway.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2014年第5期22-27,共6页
China Journal of Modern Medicine
基金
上海普陀区科委卫生系统自主创新基金(No:2010PTKW009)
上海市卫生局课题资助(No:20114059)
