转化生长因子β对椎间盘细胞Ⅰ、Ⅲ型胶原基因表达的调节作用  

The Regulating Effects of Transforming Growth Factor β(TGF-β) on Gene Expression of Collagen TypeⅠ, Ⅲin the Human Intervertebral Discs

作　　者：陈岩[1] 胡有谷[1] 吕振华[1] 

机构地区：[1]青岛大学医学院附属医院骨科,266003

出　　处：《中华骨科杂志》1999年第10期610-613,共4页Chinese Journal of Orthopaedics

基　　金：山东省卫生厅资助

摘　　要：目的 探讨转化生长因子（ｔｒａｎｓｆｏｒｍｉｎｇ ｇｒｏｗｔｈｆａｃｔｏｒβ，ＴＧＦ－β）对椎间盘Ⅰ、Ⅲ型胶原基因表达的调节作用。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、Ⅲ型胶原ｍＲＮＡ 的调节作用，采用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。结果 （１）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１ ｎｇ／ｍｌ 及１０ ｎｇ／ｍｌ 时，其调节作用分别是０ ｎｇ／ｍｌ 组的１．１８ 倍及１．３７倍；对于传代培养的纤维环细胞，其调节作用分别是０ ｎｇ／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０ ｎｇ／ｍｌ 组的１．４８ 倍及２．０３ 倍。（２） 对于原代培养的椎间盘纤维环细胞Ⅲ型胶原ｍＲＮＡ，ＴＧＦ－β１ 浓度为１ ｎｇ／ｍｌ 及１０ ｎｇ／ｍｌ 时，其调节作用分别是０ ｎｇ／ｍｌ 组的１．０８ 倍及１．２２ 倍；对于传代培养的纤维环细胞，其调节作用分别是０ ｎｇ／ｍｌ 组的１．２２ 倍及１．４０ 倍；对于传代培养的髓核细胞，其调节作用分别是０ ｎｇ／ｍｌ 组的１．２０ 倍及１．Objective To assess the regualting effects of transforming growth factor β(TGF-β) on gene expression of collagne type Ⅰ, Ⅲin the human intervertebral discs. Methods Dot blot hybridization and in situ hybridization were used to investigate the effects of TGF-β1 on collagen mRNA in confluent primary and passaged monolayer cell cultures of annulus fibrosus(AF) as well as nucleus pulposus(NP). The mean photodensitometry of hybridization membrane and cell smears as semiquantitative analysis was evaluated by VIDAS software. Results 1)Exposure to 1 ng/ml and 10 ng/ml TGF-β 1 can increase the collagen typeⅠmRNA levels by 1.18 and 1.37-fold respectively in primary cultures of AF; 1 ng/ml and 10 ng/ml TGF-β 1 can increase the collagen typeⅠmRNA levels by 1.60 and 1.84-fold respectively in passaged cultures of AF; 1 ng/ml and 10 ng/ml TGF-β1 can increase the collagen typeⅠmRNA levels by 1.48 and 2.03-fold respectively in passaged cultures of NP. 2) Exposure to 1 ng/ml and 10 ng/ml TGF-β 1 can increase the collagen type ⅢmRNA levels by 1.08 and 1.22-fold respectively in primary cultures of AF; 1 ng/ml and 10 ng/ml TGF-β1 can increase the collagen type ⅢmRNA levels by 1.22 and 1.40-fold respectively in passaged cultures of AF; 1 ng/ml and 10 ng/ml TGF-β1 can increase the collagen type ⅢmRNA levels by 1.20 and 1.43-fold respectively in passaged cultures of NP. Conclusion TGF-βcould cause a dose dependent stimulation on collagenⅠ,Ⅲgene expression in human discs, which implied that TGF-βmay exerts its great potential in the accumulation of fibrosis in the discs degeneration.

关 键 词：转化生长因子Β 胶原 基因表达 椎间盘 

分 类 号：R681.530.2[医药卫生—骨科学]