Rakopoulos M, Ikegame M, Findlay D M, Martin T J, Moseley J M
St. Vincent's Institute of Medical Research, St. Vincent's Hospital, Victoria, Australia.
Bone. 1995 Nov;17(5):447-53. doi: 10.1016/8756-3282(95)00280-8.
Cells exhibiting osteoclast characteristics of calcitonin receptors (CTRs) and tartrate-resistant acid phosphatase (TRAP) histochemistry are formed in murine bone marrow cultures treated with 1 alpha,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. We have previously demonstrated that CTR mRNA is highly expressed in these cultures. The aim of this study was to investigate homologous regulation of the CTR, and regulation of TRAP expression in osteoclast-like cells after brief treatment with salmon CT (sCT). Murine bone marrow cells were cultured in 9 cm dishes in the presence of 10 nmol/L 1,25-(OH)2D3. On day 6 of culture, when multinucleated cells were abundant, the cells were treated with 1 nmol/L sCT for 1 h. Both control and treated cells were then harvested at intervals up to 72 h posttreatment, and both CTR and TRAP mRNA levels assessed by reverse-transcription PCR (RT-PCR). In parallel cultures, cells with CTR expression detectable by autoradiography, and TRAP positivity by histochemistry, were counted. The effects of brief sCT treatment could be seen 6 h after treatment when the CTR RT-PCR product was markedly reduced. Total recovery of CTR mRNA levels had not occurred even after 72 h. Calcitonin treatment had little effect on TRAP mRNA levels. There was no difference in the numbers of multinucleated TRAP(+) osteoclast-like cells between treated and control cells. These results indicate that brief sCT treatment, while not influencing multinucleated osteoclast-like cell number, causes specific, acute reduction of CTR mRNA in bone marrow culture-derived osteoclasts. The prolonged decrease in CTR mRNA levels suggests that recovery may require new osteoclast formation, and indicates that regulation of the CTR in cells of the osteoclast lineage is different from that in nonosteoclastic cells and tissues.
在用1α,25 - 二羟基维生素D3 [1,25-(OH)2D3]处理的小鼠骨髓培养物中，会形成表现出破骨细胞特征的细胞，这些细胞具有降钙素受体（CTR）和抗酒石酸酸性磷酸酶（TRAP）组织化学特性。我们之前已经证明CTR mRNA在这些培养物中高度表达。本研究的目的是研究CTR的同源调节，以及在用鲑鱼降钙素（sCT）短暂处理后破骨细胞样细胞中TRAP表达的调节。将小鼠骨髓细胞在含有10 nmol/L 1,25-(OH)2D3的9厘米培养皿中培养。在培养的第6天，当多核细胞丰富时，用1 nmol/L sCT处理细胞1小时。然后在处理后长达72小时的间隔时间收获对照细胞和处理后的细胞，并通过逆转录聚合酶链反应（RT-PCR）评估CTR和TRAP mRNA水平。在平行培养物中，对通过放射自显影可检测到CTR表达且通过组织化学检测为TRAP阳性的细胞进行计数。短暂的sCT处理后6小时，当CTR RT-PCR产物明显减少时，就可以观察到其效果。即使在72小时后，CTR mRNA水平也没有完全恢复。降钙素处理对TRAP mRNA水平影响很小。处理后的细胞和对照细胞之间，多核TRAP(+)破骨细胞样细胞的数量没有差异。这些结果表明，短暂的sCT处理虽然不影响多核破骨细胞样细胞的数量，但会导致骨髓培养来源的破骨细胞中CTR mRNA特异性急性减少。CTR mRNA水平的持续下降表明恢复可能需要新的破骨细胞形成，并且表明破骨细胞谱系细胞中CTR的调节与非破骨细胞和组织中的调节不同。
