摘要
目的:观察Wnt经典信号通路中β-catenin在大鼠膝骨关节炎滑膜细胞胞核中的表达,并观察补肾活血方对大鼠膝骨关节炎(OA)膝关节滑膜细胞胞核中β-catenin和滑膜细胞MMP-7的调控作用。方法:采用Hulth法建立大鼠膝关节骨性关节炎模型,胶原酶消化法原代培养正常滑膜细胞与OA滑膜细胞。将所培养滑膜细胞分为正常组、OA模型组和补肾活血方组,用补肾活血方含药血清作用OA滑膜细胞48h后,采用Western Blot法检测滑膜细胞中MMP-7、胞核β-catenin的蛋白表达变化;采用酶联免疫吸附法(ELISA)检测滑膜细胞上清液中MMP-7表达变化。结果:OA滑膜细胞培养成功,WesternBlot结果显示OA滑膜细胞中MMP-7、胞核β-catenin表达均明显高于正常滑膜细胞,补肾活血方对OA滑膜细胞中MMP-7、胞核β-catenin表达有明显下调作用;ELISA结果显示OA滑膜细胞上清液中MMP-7表达明显高于正常滑膜细胞上清液,补肾活血方对其有明显下调作用。结论:①β-catenin在OA滑膜细胞胞核中表达升高,表明Wnt经典信号通路在大鼠膝骨关节炎滑膜中是激活的。②MMP-7在OA滑膜细胞和上清液中表达升高,印证了MMP-7是Wnt/β-catenin信号通路下游基因。③滑膜细胞Wnt/β-catenin信号通路的活化、MMP-7的表达升高可能是导致关节软骨降解,促进骨性关节炎形成的机制之一。④补肾活血方对Wnt/β-catenin信号通路的调控作用是其抑制软骨降解,促进软骨修复,治疗骨性关节炎的可能机制之一。
Objective ：To observe β-catenin expression of Wnt signaling pathway in rats with knee osteoarthritis,and influence of Bushen Huoxue decoction （ 补肾活血汤） on β-catenin and MMP-7 expression of synoviocytes in rats with knee osteoarthritis （OA）. Methods：Rats model with knee osteoarthritis were established by Huhh method. Primary synoviocytes and OA synoviocytes were cultured with collagenase digestion method. The cultured synoviocytes were divided into normal group, OA model group and Bus hen Huoxue decoction （补肾活血汤） group. Western blotting method was used to detect β-catenin, MMP-7 protein expression of synoviocytes after acting by Bushen Huoxue decoction （ 补肾活血汤 ） for 48 h ; ELISA method was used to detect MMP-7 expression of synovial supernatant. Results： OA synoviocytes were cultured successfully. Western blotting showed that β-catenin,MMP-7 expression in OA synoviocytes was significantly higher than normal group,Bushen Huoxue decoction （补肾活血汤） could significantly reduce β-catenin, MMP-7 expression; ELISA results showed that MMP- 7 expression of OA synovial supernatant was significantly higher than normal synoviocytes supernatant, Bushen Huoxue decoction （ 补肾活血汤） significantly regulated the level MMP-7 down. Conclusion ： （1） High expression of β-catenin in OA synoviocytes indicates that Wnt classical signal pathway is activated in rat with knee osteoarthritis; ②High expression of MMP-7 expression in OA synoviocytes confirms the MMP-7 is downstream genes of Wnt/β-eatenin signal pathway ; （3） Activation of Wnt signal pathway and increase of MMP-7 may cause degradation of articular cartilage, and promote the formation of osteoarthritis ;4Bushen Huoxue decoction（ 补肾活血汤 ） can reduce expression of MMP-7, and promote cartilage repair,which may be one of mechanisms of osteoarthritis.
出处
《中国骨伤》
CAS
2012年第9期761-765,共5页
China Journal of Orthopaedics and Traumatology
基金
上海市教委课题资助(编号:08cz041)
上海市教委高校创新团队课题资助[沪教委科(2009)6号]~~
关键词
骨关节炎
膝
补肾活血方
基质金属蛋白酶7
动物实验
Osteoarthritis, knee
Bushen Huoxue Recipe
Matrix metalloproteinase-7
Animal experimentation
