IGFBP7促进人脐带间充质干细胞的软骨分化
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国家自然科学基金面上项目(81570804,81872389)


IGFBP7 promotes human umbilical cord⁃derived mesenchymal stem cells to differentiate into chondrocytes
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    摘要:

    目的:探究胰岛素样生长因子结合蛋白7(insulin-like growth factor-binding protein 7,IGFBP7)对人脐带间充质干细胞(human umbilical cord-derived mesenchymal stem cells,hUC-MSCs)软骨分化的影响。方法:用软骨诱导培养基高密度成球培养hUC-MSCs,诱导成软骨分化,通过real-time PCR和Western blot检测软骨分化标志基因Sox9的表达,甲苯胺蓝染色检测蛋白聚糖的表达,以反映细胞外基质沉积情况;real-time PCR和Western blot检测IGFBP7在hUC-MSCs软骨分化过程中的表达情况;在hUC-MSCs中分别瞬时转染IGFBP7的表达质粒和siRNA以改变内源性IGFBP7表达水平,然后成软骨诱导,采用甲苯胺蓝染色,real-time PCR和Western blot检测IGFBP7表达变化对于hUC-MSCs成软骨分化的影响。结果:在hUC-MSCs成软骨分化过程中,IGFBP7 mRNA水平和蛋白水平表达均显著升高。与对照组相比,过表达IGFBP7的hUC-MSCs经成软骨诱导后,Sox9表达量显著升高,甲苯胺蓝染色更深,软骨分化能力更强。反之,敲低IGFBP7表达后,hUC-MSCs的软骨分化能力降低。结论:IGFBP7促进人脐带间充质干细胞成软骨分化。

    Abstract:

    Objective:This study aims to investigate the role and function of insulin-like growth factor-binding protein 7(IGFBP7)in the chondrogenic differentiation of human umbilical cord-derived mesenchymal stem cells(hUC-MSCs). Methods:Human umbilical cord-derived mesenchymal stem cells(hUC-MSCs)were cultured in high-density pellets to establish a chondrogenic differentiation model. Differences in cartilage differentiation were detected by alcian blue staining. hUC-MSCs was transfected with pCMV3-IGFBP7 or si-IGFBP7 to change the expression level of IGFBP7. The expression level of IGFBP7 was detected in chondrogenesis by real-time PCR and Western blot. The chondrogenesis-related marker gene Sox9 was determined by real-time PCR and Western blot. Results:IGFBP7 mRNA and protein expression significantly increased after induction of chondrogenic differentiation. The level of IGFBP7 in hUC-MSCs transfected with pCMV3-IGFBP7 was significantly increased,while the level of IGFBP7 in the si-IGFBP7 group was significantly reduced. Over-expression of IGFBP7 increased mRNA and protein level of Sox9 and promoted the differentiation of hUC-MSCs into chondrocyte. On the contrary,suppression of IGFBP7 inhibited chondrogenic differentiation. Conclusion:IGFBP7 was a positive regulator in the chondrogenic differentiation of hUC-MSCs.

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洪芳玲,杨家书,唐雨婷,鲍倩艺,张 明,金玉翠,李凌云,马长艳. IGFBP7促进人脐带间充质干细胞的软骨分化[J].南京医科大学学报(自然科学版),2019,(8):1101-1105

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  • 收稿日期:2019-03-17
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  • 在线发布日期: 2019-08-29
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