Stat3基因敲除小鼠成骨细胞内钙和钙通道电流变化特性的研究

Study on intracellular calcium concentration and calcium current of osteoblast in Stat3 knockout mice

目的研究Stat3基因敲除小鼠成骨细胞内钙和钙通道电流的变化特性。方法应用PCR技术对Stat3基因敲除小鼠进行鉴定;采用二次酶消化法分离、培养小鼠的原代成骨细胞;通过激光扫描共聚焦技术测定细胞内游离钙离子浓度([Ca2+]i);应用全细胞膜片钳技术记录成骨细胞膜钙通道电流(ICa)的变化。结果共聚焦结果显示Stat3基因敲除小鼠与正常小鼠相比,细胞内[Ca2+]i明显升高(P<0.01)。膜片钳结果显示,刺激电压为+10m V时,Stat3基因敲除组ICa为(-443.03±49.03)p A,与正常组[(-325.19±38.40)p A]相比,明显增加(P<0.01)。结论 Stat3基因敲除小鼠成骨细胞伴有明显的[Ca2+]i的异常,其机制可能与细胞膜上钙通道活性改变有关。

Objective To investigate the character of intracellar calcium concentration（ [Ca2 ＋]）iand calcium current（ ICa） of osteoblast in Stat3 knockout mice. Methods PCR was used to identify Stat3 knockout mice. The first generation of osteoblast was isolated and purified from the mice by sequential enzyme digestion.[Ca2 ＋]iwas detected by laser scanning confocal microscopy,and the changes of ICa were recorded by whole-cell patch-clamp technique. Results Confocal experiments showed that [Ca2 ＋]iwas markedly increased in Stat3 knockout mice,compared with that in control mice（ P〈0. 01）. Patch-clamp studies displayed that at ＋ 10 m V,the ICa was（- 443. 03 ± 49. 03） p A and（- 325. 19 ± 38. 40） p A in Stat3 knockout mice group and control group,respectively. ICa was obvious difference between two groups（ P〈0. 01）. Conclusions [Ca2 ＋]iof osteoblast is abnormal in Stat3 knockout mice, and the mechanisms are associated with the changes of electrophysiological activity of calcium channel.