转化生长因子β与椎间盘细胞Ⅰ型胶原基因调控的关系

The Regulation Effect of Transforming Growth Factor β(TGF-β) on Gene Expression of Collagen Type Ⅰ in the Human Intervertebral Discs

目的 :探讨TGF β在椎间盘Ⅰ型胶原代谢中的作用及作用机制。方法 :应用斑点杂交和原位杂交技术检测了TGF β1对椎间盘原代培养及传代培养的纤维环细胞、髓核细胞中Ⅰ型胶原mRNA的调节作用 ,采用了VIDAS软件计算杂交膜斑点及杂交涂片中细胞的平均光密度值 ,进行胶原mRNA相对定量 ,统计结果采用t检验。结果 :对于原代培养的椎间盘纤维环细胞Ⅰ型胶原mRNA ,TGF β1浓度为 1ng/ml及 10ng/ml时 ,其调节作用分别是 0ng/ml组的1.18倍及 1.3 7倍 ;对于传代培养的纤维环细胞 ,其调节作用分别是 0ng/ml组的 1.6倍及 1.84倍 ;对于传代培养的髓核细胞 ,其调节作用分别是 0ng/ml组的 1.48倍及 2 .0 3倍。结论 :TGF β可以按照剂量依赖方式正向调节椎间盘Ⅰ胶原基因表达 ,反映了TGF β与椎间盘退变过程中不断发展的纤维化过程密切相关。

Objective:To understand the characteristic and significance of the action of Transforming Growth Factor β(TGF β) on gene expression of collagen type Ⅰ in the human intervertebral discs.Method:Dotblot hybridization and in situ hybridization were used to investigate these regulation effects in confluent primary and passaged monolayer cell cultures of annulus fibrosus (AF) as well as nucleus pulposus (NP).Results:Exposure to TGF β 1 in serum free medium caused a dose dependent stimulation of collagen type ImRNA both in primary cultures of AF and in passaged cultures of AF and NP. ①1ng/ml and 10ng/ml TGF β 1 can increase the collagen type ImRNA levels by 1.18 and 1.37 fold respectively in primary cultures of AF; ②1ng/ml and 10ng/ml TGF β 1 can inerease the collagen type ImRNA levels by 1.60 and 1.84 fold respectively in passaged cultures of AF; ③lng/ml and 10ng/ml TGF β 1 can inerease the collagen type ImRNA levels by 1.48 and 2.03 fold respectively in passaged cultures of NP.Conclusion:TGF β may exerts its great potential in the accumulation of collagen type I.in the degenerative discs.