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目的通过研究ｈＰＴＨ（１～３４）对人成骨肉瘤细胞株ＳａＯＳ－２胞浆内ｃＡＭＰ、ＩＰ３、Ｃａ２＋的影响，来阐明ＰＴＨ与其膜上特异受体结合后的胞浆内主要信息传递途径。方法利用蛋白竞争结合法测定胞浆内ｃＡＭＰ，以Ｆｌｕｏ－３／ＡＭ作为荧光指示剂，激光共聚焦显微系统显示ＳａＯＳ－２内［Ｃａ２＋］ｉ的变化，离子交换层析法测定ｈＰＴＨ（１～３４）作用下胞浆内ＩＰ３的改变。结果ｈＰＴＨ（１～３４）可使ＳａＯＳ－２胞浆内ｃＡＭＰ明显升高且与其浓度呈正相关，其最佳作用时间为１０ｍｉｎ。ｈＰＴＨ（１～３４）能迅速提高ＳａＯＳ－２内［Ｃａ２＋］ｉ，而且作用３０ｓ时ＳａＯＳ－２胞浆内ＩＰ３升高即达最高峰，未发现百日咳毒素对此现象有抑制作用。结论ｃＡＭＰ及ＩＰ３／Ｃａ２＋两条信息途径均参与ｈＰＴＨ（１～３４）对成骨细胞的调节作用
Objective To study the effect of hPTH (1 ~ 34) on cAMP, IP3 and Ca2 + in the cytoplasm of human osteosarcoma cell line SaOS-2 to clarify the main intracellular informational pathway of PTH binding to its specific membrane-bound receptor. Methods The intracellular cytoplasmic cAMP was measured by protein competition and binding assay. Fluo-3 / AM was used as fluorescent indicator. The change of [Ca2 +] i in SaOS-2 was detected by laser scanning confocal microscopy. ~ 34) under the action of cytoplasmic IP3 changes. Results hPTH (1 ~ 34) could significantly increase cAMP in cytoplasm of SaOS-2 with a positive correlation with the concentration of hPTH (1 ~ 34). The optimal time of action was 10 min. hPTH (1 ~ 34) could rapidly increase [Ca2 +] i in SaOS-2, and the intracytoplasmic IP3 of SaOS-2 reached the peak at 30s. Pertussis toxin was not found to inhibit this phenomenon. Conclusions Both cAMP and IP3 / Ca2 + signaling pathways are involved in the regulation of osteoblasts by hPTH (1 ~ 34)