正常与退变椎间盘纤维环细胞相关基因的差异表达

Differential expression of intervertebral disc degeneration-related genes in normal and degenerative annulus fibrosus cells

目的:研究人正常和退变椎间盘纤维环细胞椎间盘退变相关基因表达量的差异,探讨这些相关基因与椎间盘退变的关系,筛选明显差异基因作为退变纤维环细胞体外刺激因子,逆转椎间盘退变。方法:实时荧光定量PCR(real-time qRT-PCR,SYBR Green)技术检测人正常和退变纤维环细胞TGF-β、IGF-1、bFGF、IL-1、TIMP-1、COL1A1、BMP-14、PDGF、aggrecan、COL2A1、SOX 9、BMP-2、iNOS、EGF、MMP3、BMP-4和BMP-7mRNA表达水平。结果:与正常纤维环细胞相比,退变纤维环细胞TGF-β、IL-1、PDGF和IGF-1 mRNA表达量降低(P<0.01,P<0.05);bFGF、TIMP-1、BMP-14和COL1A1 mRNA表达升量高(P<0.01);正常纤维环细胞aggrecan和BMP-2 mRNA高表达,退变纤维环细胞两者表达阴性;COL2A1、iNOS和MMP3在正常纤维环细胞mR-NA有表达,在退变纤维环细胞表达阴性;EGF在正常纤维环细胞mRNA表达阴性,而在退变纤维环细胞有表达。Sox-9、BMP-4和BMP-7在正常和退变纤维环细胞mRNA表达均为阴性。结论:椎间盘纤维环退变与TGF-β、IL-1、PDGF、IGF-1、aggrecan和BMP-2 mRNA低表达,bFGF、TIMP-1、BMP-14和COL1A1 mRNA高表达相关;BMP-2、TGF-β、PDGF和IGF-1可作为退变纤维环细胞体外刺激因子,用于逆转椎间盘退变的研究;bFGF、TIMP-1和BMP-14可能是椎间盘纤维环细胞退变的标志物。

AIM： To study the differential expression levels of 17 intervertebral disc degeneration-related genes in human normal and degenerative annulus fibrosus（AF） cells and to explore the relationship of these genes and intervertebral disc degeneration.METHODS： The mRNA expression levels of TGF-β,IGF-1,bFGF,IL-1,TIMP-1,COL1A1,COL2A1,AGGRECAN,MMP3,iNOS,BMP-2,BMP-7,BMP-14,BMP-4,EGF,PDGF and SOX9 were detected by real-time quantitative RT-PCR.RESULTS： The mRNA expression levels of TGF-β,IL-1,PDGF and IGF-1 were lower and bFGF,TIMP-1,BMP-14 and COL1A1 were higher in degenerative AF cells than those in normal AF cells.The mRNA expression levels of aggrecan and BMP-2 were high in normal AF cells but negative in degenerative AF cells.The mRNA expression levels of COL2A1,iNOS and MMP3 were positive in normal AF cells but negative in degenerative AF cells.The mRNA expression of EGF was negative in normal AF cells but positive in degenerative AF cells.The mRNA expression levels of SOX-9,BMP-4 and BMP-7 were negative both in normal AF cells and in degenerative AF cells.CONCLUSION： According to the gene expression profiles,the degeneration of intervertebral disc is associated with the low gene expression of TGF-β,IL-1,PDGF,IGF-1,aggrecan and BMP-2,and the high gene expression of bFGF,TIMP-1,BMP-14 and COL1A1.Therefore,BMP-2,TGF-β,PDGF and IGF-1 may be as stimulating factors for degenerative AF cells to reverse the intervertebral disc degeneration.