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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 investigate the role of TGFβ in type Ⅰ collagen metabolism of disc and its mechanism. Methods: Dot blotting and in situ hybridization were used to detect the regulation of type Ⅰ collagen mRNA in fibroblasts and nucleus pulposus cells by primary culture of disc and subcultured by TGF β1. VIDAS software was used to calculate the hybridization spots The average optical density of cells in the cells, the relative quantitative collagen mRNA, statistical results using t test. Results: For typeⅠcollagen mRNA of primary cultured fibrocystic ring cells, when TGFβ1concentration was1ng / ml and10ng / ml, the regulatory effect was1.18times and1.37times respectively in0ng / ml group; For subcultured The regulation of fibroblast cells was 1.6 times and 1.84 times that of 0ng / ml group, respectively. The regulation effect on subcultured nucleus pulposus cells was 1.48 times and 2.03 times that of 0ng / ml group respectively. CONCLUSION: TGFβ can positively regulate collagen Ⅰ gene expression in intervertebral disc Ⅰ in a dose-dependent manner, which indicates that TGFβ is closely related to the development of fibrosis in the process of intervertebral disc degeneration.