Thiébaud D, Guenther H L, Porret A, Burckhardt P, Fleisch H, Hofstetter W
Department of Internal Medicine, University Hospital, Lausanne, Switzerland.
J Bone Miner Res. 1994 Sep;9(9):1347-54. doi: 10.1002/jbmr.5650090905.
Growth factors, such as transforming growth factor beta (TGF-beta) and insulin-like growth factors (IGF) I and II, have been shown to exert anabolic effects on bone cells in vitro. Hormones, such as PTH and probably insulin and growth hormone, were recently shown to stimulate bone formation in vivo as well. The aim of the present study was to assess by northern blots, which were quantitated by densitometry, the effects of these anabolic growth factors and hormones in two osteogenic cell populations: CRP 10/30 cells, a clonal cell population derived from primary rat calvarial cells, and IRC 10/30-myc cells, which were established from CRP 10/30 by immortalization. Transcripts for alpha 1(I) collagen, biglycan, osteonectin, osteopontin, and osteocalcin were detected in both cell populations, which is consistent with the phenotype expressed by mature osteoblasts. There were no difference in the basal expression of bone matrix mRNAs between the two cell populations. PTH increased alpha 1(I) collagen mRNA levels in both osteoblastic cells but had no effect on the biglycan transcripts. Neither insulin nor growth hormone affected mRNA levels of either matrix protein after 24 h exposure. All three growth factors, TGF-beta, IGF-I, and IGF-II, increased alpha 1(I) collagen transcripts in a time- and dose-dependent manner in both cell populations. Biglycan mRNA levels were enhanced in both osteoblastic lines only by IGF-I and IGF-II, but not TGF-beta.(ABSTRACT TRUNCATED AT 250 WORDS)
生长因子,如转化生长因子β(TGF-β)以及胰岛素样生长因子(IGF)-I和IGF-II,已被证明在体外对骨细胞具有合成代谢作用。激素,如甲状旁腺激素(PTH),可能还有胰岛素和生长激素,最近也被证明在体内能刺激骨形成。本研究的目的是通过经光密度测定进行定量的Northern印迹法,评估这些合成代谢生长因子和激素对两种成骨细胞群体的影响:CRP 10/30细胞,一种源自原代大鼠颅骨细胞的克隆细胞群体;以及IRC 10/30-myc细胞,它是通过CRP 10/30永生化建立的。在这两种细胞群体中均检测到α1(I)胶原蛋白、双糖链蛋白聚糖、骨连接蛋白、骨桥蛋白和骨钙素的转录本,这与成熟成骨细胞表达的表型一致。这两种细胞群体之间骨基质mRNA的基础表达没有差异。PTH使两种成骨细胞中的α1(I)胶原蛋白mRNA水平升高,但对双糖链蛋白聚糖转录本没有影响。暴露24小时后,胰岛素和生长激素均未影响任何一种基质蛋白的mRNA水平。所有三种生长因子,即TGF-β、IGF-I和IGF-II,在两种细胞群体中均以时间和剂量依赖性方式增加α1(I)胶原蛋白转录本。仅IGF-I和IGF-II增强了两种成骨细胞系中的双糖链蛋白聚糖mRNA水平,而TGF-β没有。(摘要截短于250字)