The effects of stimulators of intracellular cyclic AMP on rat and chick osteoclasts in vitro: validation of a simplified light microscope assay of bone resorption.
The effects of stimulators of intracellular cyclic AMP on rat and chick osteoclasts in vitro: validation of a simplified light microscope assay of bone resorption.
复制标题
细胞内环磷酸腺苷刺激剂对体外大鼠和鸡破骨细胞的影响：骨吸收简化光学显微镜测定的验证。
DOI：
10.1016/8756-3282(90)90089-h
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发表时间：
1990
期刊：
影响因子：
4.1
通讯作者：
Dempster,DW
中科院分区：
文献类型：
--
作者：
Murrills,RJ;Dempster,DW
本研究的目的是调查是否存在一个环AMP介导的抑制机制，在胚胎鸡破骨细胞和扩展数据牵连环AMP在新生大鼠破骨细胞的抑制。二丁酰环腺苷酸（（Bu）2cAMP）（5 × 10− 4 M及以上）和异丁基甲基黄嘌呤（IBMX）（10− 4 M及以上）减少了鸡破骨细胞在24小时内在失活的牛皮质骨切片中形成的凹坑数量。毛喉素（FSK）对鸡破骨细胞的作用是双相的，10 - 5 M产生微弱的和可变的小窝数量减少，而10 - 6 M和10 - 7 M刺激吸收。单独使用FSK（10− 5 M）和（Bu）2cAMP（3 × 10− 4 M）不会产生一致的显著效应，但当联合使用时，会产生协同和高度显著的小坑数量减少，这意味着这些药物通过共同的机制起作用，可能是环AMP。刺激剂量的FSK与破骨细胞数量增加有关，表明环AMP参与破骨细胞的形成。在使用新生大鼠破骨细胞的对比实验中，（Bu）2cAMP（10− 4 M及以上）、IBMX（10− 3 M）和FSK（10− 7 M及以上）都减少了凹陷的数量。这些药物的强抑制剂量导致鸡破骨细胞收缩成半球形;（Bu）2cAMP和FSK可使大鼠破骨细胞收缩成星状，但IBMX则不然。我们的研究结果牵连环磷酸腺苷在大鼠和鸡破骨细胞的抑制，并表明，在光学显微镜下的小坑计数是一种有效的方法来分析解聚破骨细胞吸收测定。
The aim of this study was to investigate whether a cyclic AMP-mediated inhibitory mechanism is present in embryonic chick osteoclasts and to extend data implicating cyclic AMP in the inhibition of neonatal rat osteoclasts. Dibutyryl cyclic AMP ((Bu)2cAMP) (5 × 10−4M and above) and isobutylmethylxanthine (IBMX) (10−4M and above) reduced the number of pits made in slices of devitalized bovine cortical bone by chick osteoclasts over 24 h. The effect of forskolin (FSK) on chick osteoclasts was biphasic, 10−5M producing a weak and variable reduction in pit number while 10−6M and 10−7M stimulated resorption. Doses of FSK (10−5M) and (Bu)2cAMP (3 × 10−4M), which individually produced no consistent significant effect, produced a synergistic and highly significant reduction in pit number when used in combination, implying that these agents were acting through a common mechanism, presumably cyclic AMP. Stimulatory doses of FSK were associated with increased osteoclast numbers, implicating cyclic AMP in the formation of osteoclasts. In comparative experiments using neonatal rat osteoclasts, (Bu)2cAMP (10−4M and above), IBMX (10−3M) and FSK (10−7M and above) all reduced the number of pits excavated. Strongly inhibitory doses of these agents caused contraction of chick osteoclasts into a hemispherical shape; contraction of rat osteoclasts into a stellate shape occurred with (Bu)2cAMP and FSK, but not with IBMX. Our results implicate cyclic AMP in the inhibition of both rat and chick osteoclasts, and show that pit counting in the light microscope is a valid method of analyzing the disaggregated osteoclast resorption assay.