bFGF对人退变椎间盘髓核细胞影响的临床研究

Clinic research of the effect of bFGF on human degenerated nucleus pulposus cells

目的:探讨bFGF对人退变髓核细胞合成细胞外基质的影响,观察退变髓核细胞基因表达量高于正常髓核细胞的生长刺激因子对退变髓核细胞的影响。方法:分离、培养人退变椎间盘髓核细胞,取第2代髓核细胞,随机将退变椎间盘髓核细胞分为5组。A组:加入100μg/L bFGF;B组:加入200μg/L bFGF;C组:500μg/L bFGF;D组:1 000μg/LbFGF,E组:对照组,不加干扰因素。通过对试验组和对照组髓核细胞测定细胞II型胶原和糖胺多糖的mRNA表达;检测细胞外基质II型胶原和糖胺多糖表达水平。结果:bFGF刺激退变髓核细胞,II型胶原、糖胺多糖mRNA,细胞外液Ⅱ型胶原和糖胺多糖含量在第7天较对照组增加明显(P<0.05)。14 d、21 d II型胶原、糖胺多糖mRNA明显低于对照组(P<0.05)。但细胞外液Ⅱ型胶原和糖胺多糖含量在14 d、21 d两个时间点仍高于正常组(P<0.05)。结论:根据本研究结果结合文献调研,bFGF在椎间盘髓核细胞中存在着双重效应(促进或改善椎间盘退变)。

AIM： To determine the effect of bFGF on extracellular matrix and gene expression on the human degenerated nucleus pulposus cells.METHODS： The degenerated intervertebral disc cells were divided into 5 groups.A group： adding 100 μg/L bFGF,B group： Adding 200 μg/L bFGF,C group： 500 μg/L bFGF,D group： 1000 μg/L bFGF,E groups： control group,without interfering factors.The type II collagen and GAG mRNA expression were test,type Ⅱ collagen content and glycosaminoglycan content of the supernatant were test.RESULTS： bFGF stimulated degenerated nucleus pulposus cells,collagen II,GAG mRNA,collagen type II and glycosaminoglycan expression of the extracellular matrix were increased significantly on the 7 days compared with the control group（P〈0.05）.type II collagen and GAG mRNA were significantly lower than the control group on 14 days,21 days（P〈0.05）.However,collagen type II and glycosaminoglycan expression of the extracellular matrix on 14 days,21 days were still higher than normal（P〈0.05）.CONCLUSION： bFGF increase GAG and type II collagen mRNA expression,increased extracellular matrix type II collagen and GAG expression in short-term.