Jongen J W, Bos M P, van der Meer J M, Herrmann-Erlee M P
Laboratory of Cell Biology and Histology, University of Leiden, The Netherlands.
J Cell Physiol. 1993 Apr;155(1):36-43. doi: 10.1002/jcp.1041550106.
We studied the effects of parathyroid hormone (PTH) on two markers of the osteoblast phenotype: alkaline phosphatase (AP) (activity and mRNA) and cyclic adenosine monophosphate (cAMP) accumulation. Osteoblast-like cells derived from fetal rat (ROB) and mouse (MOB) calvariae were isolated by collagenase treatment. Cells were cultured in alpha-Minimal Essential Medium (MEM) with 2% fetal calf serum (FCS) for 4 days. In ROB and MOB bPTH(1-34) induced a fast increase (up to 5 minutes) in cAMP accumulation. When equal amounts of cells were seeded, the cAMP accumulation was higher in MOB than in ROB. No difference in basal AP activity was observed between ROB and MOB. When bpTH (1-34) was added to ROB for the last 24 or 48 hr, AP activity decreased dose dependently. However, MOB treated with bPTH(1-34) for the last 24 or 48 hours showed an increase of AP activity. Basal AP activity was positively correlated with the seeding density of ROB and MOB cultures. Basal AP activity influenced the degree of inhibition (ROB) or stimulation (MOB) after incubation with bPTH(1-34).
我们研究了甲状旁腺激素(PTH)对成骨细胞表型的两个标志物的影响:碱性磷酸酶(AP)(活性和mRNA)以及环磷酸腺苷(cAMP)积累。通过胶原酶处理从胎鼠(ROB)和小鼠(MOB)颅骨分离出成骨样细胞。细胞在含2%胎牛血清(FCS)的α-最低必需培养基(MEM)中培养4天。在ROB和MOB中,bPTH(1-34)诱导cAMP积累快速增加(长达5分钟)。当接种等量细胞时,MOB中的cAMP积累高于ROB。在ROB和MOB之间未观察到基础AP活性的差异。当在最后24或48小时向ROB中添加bPTH(1-34)时,AP活性呈剂量依赖性降低。然而,在最后24或48小时用bPTH(1-34)处理的MOB显示AP活性增加。基础AP活性与ROB和MOB培养物的接种密度呈正相关。基础AP活性影响与bPTH(1-34)孵育后的抑制程度(ROB)或刺激程度(MOB)。