敲低NIPBL基因对TGF-β1与IGF-1联合诱导骨髓间充质干细胞成软骨分化的影响

Effect of Knockdown of NIPBL Gene on Chondrogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Induced by Combination of TGF-β1 and IGF-1

目的:探讨敲低NIPBL基因对TGF-β1与IGF-1联合诱导骨髓间充质干细胞成软骨分化的影响。方法:将慢病毒转染的NIPBL shRNA骨髓间充质干细胞分为TGF-β1组、TGF-β1+IGF-1组、NIPBL-空白对照组,并设立未转染的NIPBL+组,对上述四组细胞进行成软骨诱导分化;在培养7、14、21天时应用Western blot技术分别检测Sox-9、Collagen-Ⅱ的蛋白表达水平。结果:(1)在成软骨诱导的各阶段,Sox-9、Collagen-Ⅱ的蛋白表达量均为TGF-β1+IGF-1组高于TGF-β1组,NIPBL+组高于TGF-β1组和NIPBL-对照组(P<0.05);(2)在诱导7、21天时,Sox-9基因的蛋白表达量TGF-β1+IGF-1组高于NIPBL-对照组(P<0.05);(3)在各诱导阶段,TGF-β1+IGF-1组和NIPBL+组间差异无统计学意义。结论:敲低NIPBL基因导致TGF-β1与IGF-1联合诱导骨髓间充质干细胞成软骨分化的能力降低。

Objective:To investigate the effect of knockdown of NIPBL gene on TGF-β1 and IGF-1 combined to induce chondrogenic differentiation of bone marrow mesenchymal stem cells.Methods:Lentiviral-transfected NIPBL shRNA BMSCs were divided into TGF-β1 group,TGF-β1+IGF-1 group,NIPBL-blank control group,and an untransfected NIPBL+control group,and the above four groups of cells were subjected to chondrogenic differentiation induction.The protein expression levels of Sox-9 and Collagen-Ⅱwere detected by Western blot technique at 7,14 and 21 days of culture.Results:(1)At each stage of chondrogenesis induction,the protein expression of Sox-9 and Collagen-Ⅱwere higher in the TGF-β1+IGF-1 group than in the TGF-β1 group,and higher in the NIPBL+control group than in the TGF-β1 and NIPBL-control groups(P<0.05).(2)At 7 and 21 days of induction,the protein expression of Sox-9 gene was higher in the TGF-β1+IGF-1 group than in the NIPBL-control group(P<0.05).(3)At each induction stage,there was no statistical difference between the TGF-β1+IGF-1 and the NIPBL+control group.Conclusion:Knockdown of NIPBL gene TGF-β1 combined with IGF-1 has reduced ability to induce chondrogenic differentiation of bone marrow mesenchymal stem cells.