不同浓度TGF-β1对人髓核细胞外基质成分基因表达的调控作用比较

Comparative study on the regulation effects of TGF-β1 at different concentration in vitro on cellular aggrecan,collagen Ⅱ and SOX-9 gene expression of human nucleus pulposus

目的研究在体外培养条件下,不同浓度转化生长因子-β1(TGF-β1)对人髓核细胞聚集蛋白聚糖(Aggrecan)、Ⅱ型胶原及SOX-9基因表达的调控作用。方法以传2代人髓核细胞为研究对象,分别以0.1μg/L、1.0μg/L、10μg/L、100μg/L四种浓度的TGF-β1为培养液,设不含生长因子培养液为对照组,应用逆转录聚合酶链反应(RT-PCR)检测Aggrecan、Ⅱ型胶原和SOX-9 m RNA表达情况。结果 0.1μg/L TGF-β1组与对照组相比,无显著性差异。1μg/L、10μg/L和100μg/L TGF-β1组在促进Aggrecan、Ⅱ型胶原和SOX-9基因表达方面作用显著,差异均具统计学意义。其中,10μg/L组促进细胞m RNA表达作用最明显。结论 TGF-β1能够促进髓核细胞Aggrecan、Ⅱ型胶原和SOX-9基因的表达,提示TGF-β1有可能在椎间盘退行性疾患的预防和治疗中发挥重要作用。

Objective To investigate the regulation effects of TGF-β1 on aggrecan,collagen II and SOX-9 gene expression of human nucleus pulposus at different concentration in vitro. Methods Passage 2 nucleus pulposus cells were cultured in the medium with 10%FBS and at different concentrations （TGF-β1 as 0.1 μg/L, 1.0μg/L,10 μg/L,100 μg/L）. The medium without TGF-β1 was taken as control group. The mRNA expression of aggrecan,eollagen Ⅱ and SOX-9 gene was assayed by RT-PCR. Results The 0.1 μg/L TGF-β1 group hadn＇t significant difference compared with the control group. The 1 μg/L,10 μg/L and 100 μg/L groups,promoted mRNA expression of aggrecan,collagen Ⅱ and SOX-9 gene compared with the control group,the difference had statistical significance. Conclusion TGF-β1 is efficient to promote aggrecan,collagen ]I and SOX-9 gene expression of human nucleus pulposus which might be a future therapeutic potential in the treatment of intervertebral disc degeneration.