肾上腺髓质素对低氧时肺成纤维细胞内钙离子动态变化的影响

Effects of adrenomedullin on concentration change of intracellular calciumion([Ca^(2+)]_i) in human fetal lung fibroblasts induced by hypoxia

目的动态观察低氧(2%O2)对培养的人胚肺成纤维细胞内游离钙离子浓度的影响及肾上腺髓质素预处理对其作用。方法 1、采用体外细胞培养方法,培养并鉴定人胚肺成纤维细胞;2、建立人胚肺成纤维细胞低氧(2%O2)模型;3、采用激光扫描共聚焦显微镜技术动态观测低氧时成纤维细胞内游离钙离子浓度的变化及肾上腺髓质素对其影响。结果低氧促进成纤维细胞内游离钙离子浓度升高。与常氧组比较,低氧后成纤维细胞内游离钙离子浓度增加,标准荧光值峰值上升了70%(P<0.01)。肾上腺髓质素预处理成纤维细胞后,低氧引起的成纤维细胞内游离钙离子浓度升高受到抑制,标准荧光值峰值上升了40%(P<0.01),持续时间缩短了20s。结论肾上腺髓质素可以抑制低氧引起的人胚肺成纤维细胞内游离钙离子浓度增加,提示这可能是其发挥调节成纤维细胞功能的作用机制之一。

Objective To observe the effects of adrenomedullin （AM） on the changes in the concentration of intracellular caleiumion （[Ca^2＋]1） in human fetal lung fibroblasts induced by hypoxia. Methods Human fetal lung fibroblasts were cultured by outgrowth in vitro. Fibroblasts grown on a Petri dish were placed on the stage of an inverted microscope. The fluorescence signals of five representative fibroblasts were measured with a confocal laser scanning system. Results Compared with standard oxygen levels, low oxygen level （2% O2） increased [Ca^2＋]1 in human lung fibroblasts by 70%. After pretreatment with AM （10-7mool/L） in normal culture medium for 5 min, low oxygen level （2%O2） increased [Ca^2＋]1in human lung fibroblasts by 40%. Conclusion The effects of hypoxia on [Ca^2＋]i were significantly inhibited by pretreatment with AM, suggesting that the Ca^2＋ influx response to hypoxia may be blocked by AM in cultured human fetal lung fibroblasts.