基础医学与临床 ›› 2016, Vol. 36 ›› Issue (12): 1624-1629.

• 研究论文 • 上一篇    下一篇

Wnt/β-catenin信号通路在GDF-5和Dex促进rBMSCs体外成软骨分化中的作用

汪建样1,殷嫦嫦2,邬亚华2,严晨3,王子瑶1,殷明3   

  1. 1. 南昌大学江西医学院
    2. 九江学院
    3. 南昌大学第二附属医院
  • 收稿日期:2015-07-13 修回日期:2015-11-12 出版日期:2016-12-05 发布日期:2016-11-29
  • 通讯作者: 殷嫦嫦 E-mail:113861794@qq.com
  • 基金资助:
    国家自然科学基金

Wnt / β-catenin signaling pathway promotes cartilage differentiation of BMSCs

  • Received:2015-07-13 Revised:2015-11-12 Online:2016-12-05 Published:2016-11-29

摘要: 目的 利用生长分化因子(GDF-5)联合地塞米松(Dex)诱导大鼠BMSCs成软骨分化,并探讨Wnt/β-catenin信号通路在其中的作用。方法:用全骨髓贴壁法分离培养SD大鼠BMSCs,取P3代细胞随机分成6组:BMSCs组、BMSCs+Dex组、BMSCs+GDF-5组、BMSCs+GDF-5+Dex组、BMSCs+ GDF-5+ Dex+SB216763组和BMSCs+GDF-5+Dex+ XAV-939组。连续诱导培养14d,倒置相差显微镜观察细胞形态,Alcian blue染色检测细胞的蛋白聚糖,RT-PCR检测成软骨分化相关基因Aggrecan、Col2、Sox9、Col1,Wnt/β-catenin信号通路相关基因Dvl1、Gsk3β、β-catenin mRNA表达,Western blot检测Ⅱ型胶原蛋白和β-catenin蛋白表达。结果:与BMSCs组相比,BMSCs+GDF-5+Dex组能够使诱导的细胞蛋白聚糖分泌明显增加;Aggrecan、Col2、Sox9基因表达明显增加(P<0.05);Ⅱ型胶原蛋白表达增加(P<0.05);β-catenin mRNA及蛋白表达水平也增加(P<0.05)。与BMSCs+GDF-5+Dex组相比,添加SB216763组蛋白聚糖,Ⅱ型胶原基因和蛋白表达增加(P<0.05);相反,添加XAV-939组相应的成软骨分化基因及蛋白表达下降(P<0.05)。结论:GDF-5联合地塞米松(Dex)可诱导大鼠BMSCs成软骨分化,Wnt/β-catenin信号通路可能在其中发挥促进作用。

关键词: 关键词:Wnt/β-catenin , BMSCs, 成软骨分化, GDF-5

Abstract: Objective Using Growth differentiation factor GDF-5 in combination with dexamethasone (Dex) to induce BMSCs into cartilage differentiation and to explore the role of Wnt/β-catenin signaling pathway in cartilage differentiation of rBMSCs.Methods BMSCs were collected from Sprague Dawley rats.The Morphology was observed under inverted microscope .rBMSCs at passage 3 were divided into 6 groups: BMSCs group,BMSCs+Dex group,BMSCs+GDF-5 group,BMSCs+ GDF-5+Dex group,BMSCs+ GDF-5+ Dex+SB216763 group,BMSCs+GDF-5+Dex+ XAV-939 group and induced for 14 days.Alcian blue stain was used to show the changes of proteoglycans ; RT-PCR was used to detecte the mRNA expression of Aggrecan,COL2,Sox9,COL1,Dvl1,Gsk3β and β-catenin.The protein expression levels of COL2 and β-catenin were detected by Western blot. Results Compared with the BMSCs group, BMSCs+GDF-5+Dex group showed deeper proteoglycan staining; Aggrecan, COL2, Sox9 and the protein expression levels of COL2 were significantly increased;β-catenin mRNA and protein expression levels also increased. Compared with the BMSCs + GDF-5 + Dex group,in adding SB216763 group proteoglycans, collagen type Ⅱ gene and protein expression were increased significantly (P <0.05); On the contrary,in adding XAV-939 group cartilage differentiation gene and protein expression were decreased (P <0.05).Conclusion GDF-5 in combination with dexamethasone (Dex) can induce differentiation of rat BMSCs into cartilage, Wnt/β-catenin signaling pathway may play a promoter role.

Key words: Key words: Wnt/β-catenin , BMSCs, Chondrogenic differentiation, GDF-5

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