维生素D_3刺激髁突软骨细胞内钙释放及机械压力对其影响的研究  

Intracellular Calcium Ion Release Stimulated by 1,25(OH)_2D_3 and Influenced with Mechanical Pressure in Mandibular Condylar Chondrocytes of Rabbit

作　　者：张旻 王美青[1] 王景杰[1] 

机构地区：[1]第四军医大学口腔医学院解剖生理教研室,陕西西安710032

出　　处：《华西口腔医学杂志》2004年第1期69-72,共4页West China Journal of Stomatology

基　　金：国家自然科学基金资助项目 (30 0 0 0 0 35)

摘　　要：目的　探讨维生素D3(1,2 5 (OH) 2 D3)刺激体外培养的兔髁突软骨细胞内钙离子的释放通道及机械压力对其影响。方法　消化法培养新西兰白兔髁突软骨细胞 ,分别进行肝素 (2 0g L)、普鲁卡因 (1g L)钙通道阻断处理 ,及用自行设计制作的可控液压细胞加载装置分别进行 90kPa 6 0min和 90kPa 36 0min的加压处理 ,经钙荧光指示剂Fluo_3负载后 ,用激光扫描共聚焦显微镜测定 10 _8mol L 1,2 5 (OH) 2 D310 0 μL刺激前后髁突软骨细胞内钙随时间变化情况 ,同时设有对照组。结果　 1,2 5 (OH) 2 D3刺激后对照组细胞内荧光强度随时间明显升高 ;普鲁卡因处理组变化与其相似 ,而肝素处理组在刺激前后胞内荧光强度无明显变化 ;90kPa加压 6 0min处理组在接受 1,2 5 (OH) 2 D3刺激后胞内荧光强度也随时间明显升高 ,且其升高幅度显著高于对照组 ,而 90kPa加压 36 0min处理组在刺激后胞内荧光强度虽也有升高 ,但与对照组无显著差异 ,且在记录末期出现下降趋势。结论　 1,2 5 (OH) 2 D3能刺激髁突软骨细胞内三磷酸肌醇受体 (IP3R)钙释放通道开放 ,使细胞内钙离子水平显著升高 ;Objective To investigate the intracellular calcium ion release and the system of calcium channel by 1,25(OH)-2D-3 stimulus, and the effect of mechanical pressure on it in rabbit mandibular condylar chondrocytes (MCC) in vitro. Methods In vitro cultured MCC from two-week-old New Zealand rabbits were incubated under 20 g/L heparin, 1 g/L procaine, continuous pressure of 90 kPa for 60 min and 360 min in a hydraulic pressure controlled cellular strain unit. With the Fluo-3/AM probe loaded, 1,25(OH)-2D-3 was added to the medium and then the intracellular calcium level was detected by a laser confocal scanning microscope.Results Intracellular calcium concentration increased in MCC treated with 1,25(OH)-2D-3, 1,25(OH)-2D-3 and procaine, while it didn′t change in heparin treated group. Calcium in group under continuous pressure of 90 kPa for 60 min was also increased, even higher than the group stimulated only with 1,25(OH)-2D-3. Intracellular calcium in group treated with continuous pressure of 90 kPa for 360 min showed no significant difference compared to the control and even decreased at the end of the recording period. Conclusion 1,25(OH)-2D-3 could stimulate the intracellular calcium release channel of inositol triphosphate (IP3) receptor open in MCC in vitro and increases the level of intracellular calcium concentration. Pretreatment of definite mechanical pressure could modulate the sensitivity of IP3 channel to 1,25(OH)-2D-3 stimulus.

关 键 词：维生素D3 髁突软骨细胞 钙释放 机械压力 激光扫描共聚焦显微镜 

分 类 号：R780.2[医药卫生—口腔医学]