【摘要】 目的探讨肝细胞生长因子(HGF)对体外培养的人退变髓核细胞的生物学作用。方法收集人退变髓核组织标本,分离培养髓核细胞;免疫组化染色观察HGF受体在髓核细胞中表达;设立不同浓度HGF组(0ng/ml,5ng/ml,50ng/ml,500ng/ml),采用MTT法测细胞生长曲线,筛选HGF最佳作用浓度;选取第3代髓核细胞,实验分HGF组、HGF+胰岛素样生长因子(IGF)组、IGF组、对照组,RT-PCR检测Ⅱ型胶原、蛋白多糖、SOX9mRNA表达;流式细胞术测定细胞增殖的周期。结果免疫组化显示髓核细胞表达HGF受体;MTT法提示不同浓度HGF对体外培养的人退变髓核细胞均有明显的促增殖作用,与对照组相比差异均有统计学意义,50ng/ml是实验中HGF的最佳作用浓度;HGF、HGF+IGF、IGF刺激后髓核细胞进入增殖期比例增加,同时促进髓核细胞Ⅱ型胶原、蛋白多糖、SOX9mRNA表达量增加,其中HGF+IGF效果更为显著。结论人退变髓核细胞存在HGF受体;HGF对体外培养的人退变的髓核细胞有促增殖作用,可以促进细胞外基质表达;HGF与IGF联合作用效果优于两者单独作用。更多还原

【Abstract】 Objective To explore the effect of hepatocyte growth factor (HGF) on the biological activity of the human degeneration nucleus pulposus cells cultured in vitro.Methods The human nucleus pulposus were collected,and the nucleus pulposus cells were isolated and cultured.To observe HGF receptor expression in nucleus pulposus cells by immunohistochemical staining.The experimental groups were cultured with 0,5,50,500 ng/ml HGF,the optimal concentration was selected by MTT.The 3rd generation of nucleus pulposus cells were selected ard divided into HGF,HGF+IG,IGF and control group,reverse transcription-polymerase chain reaction (RT-PCR) was used to determine levels of type Ⅱ collagen,proteoglycan and SOX9;flow cytometer were used to detect cell cycle.Results Nucleus pulposus cells expressed HGF receptor.HGF could significantly promote the proliferation of the human degeneration nucleus pulposus cells,which had a significant different compared with the control group.Various concentrations of HGF were effective to promote the nucleus pulposus cells proliferation and the optimal HGF concentration was 50 ng/ml.The human degeneration nucleus pulposus cells under HGF,IGF and HGF+IGF had shown to keep typeⅡ collagen,proteoglycan and SOX9 expression positive and increasing,and the proportion of cells into the proliferative phase increased compared with control group,which HGF+IGF was more effective.Conclusions Nucleus pulposus cells expressed HGF receptor.HGF can promote the proliferation of the human degeneration nucleus pulposus cells cultured in vitro,enhance the expression of extracellular matrix.The combined effects of HGF and IGF are better than alone.更多还原