Turner R T, Spelsberg T C
Department of Orthopedic Surgery, Mayo Clinic, Rochester, Minnesota 55905.
Am J Physiol. 1991 Sep;261(3 Pt 1):E348-53. doi: 10.1152/ajpendo.1991.261.3.E348.
This report describes the relationship between bone formation and mRNA levels for selected bone proteins. Dynamic bone histomorphometry was used to measure bone formation in tibial periosteum of male rats from weanling (3 wk) to 52 wk old. Northern blot analysis of freshly isolated periosteal cells from the long bones was used to determine steady-state mRNA levels for the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAP), the bone matrix proteins osteocalcin (BGP), and prepro-alpha-2 (I) chain of type 1 precollagen (collagen), the osteoblast marker enzyme alkaline phosphatase (AP), and the osteoblast-derived signaling factor (growth factor) transforming growth factor-beta (TGF-beta). Radial growth at the tibial diaphysis achieved a maximum value in 8-wk-old rats and decreased progressively with age thereafter. This age-related decrease in the radial growth rate was initially due to reduced osteoblast activity; however, in older rats (greater than 17 wk old) reduced osteoblast number contributed to the decrease in bone formation. There was a strong correlation between the steady-state mRNA level for collagen and the periosteal bone formation rate. In contrast, the mRNA levels for the other bone proteins were more weakly correlated (TGF-beta and AP) or not correlated (BGP). These results suggest that the decreased bone matrix synthesis by periosteal cells in long bones of maturing rats is due to decreased expression of genes for bone matrix proteins.
本报告描述了特定骨蛋白的骨形成与mRNA水平之间的关系。采用动态骨组织形态计量学方法测量雄性大鼠从断奶(3周)至52周龄胫骨骨膜的骨形成。运用Northern印迹分析法,对从长骨新鲜分离的骨膜细胞进行检测,以确定糖酵解酶甘油醛-3-磷酸脱氢酶(GAP)、骨基质蛋白骨钙素(BGP)、Ⅰ型前胶原前体α-2(I)链(胶原蛋白)、成骨细胞标记酶碱性磷酸酶(AP)以及成骨细胞衍生的信号因子(生长因子)转化生长因子-β(TGF-β)的稳态mRNA水平。胫骨骨干的径向生长在8周龄大鼠时达到最大值,此后随年龄增长逐渐下降。这种与年龄相关的径向生长速率下降最初是由于成骨细胞活性降低;然而,在老年大鼠(大于17周龄)中,成骨细胞数量减少也导致了骨形成的下降。胶原蛋白的稳态mRNA水平与骨膜骨形成速率之间存在很强的相关性。相比之下,其他骨蛋白的mRNA水平相关性较弱(TGF-β和AP)或无相关性(BGP)。这些结果表明,成熟大鼠长骨中骨膜细胞骨基质合成的减少是由于骨基质蛋白基因表达的降低。