豚鼠膀胱Cajal样间质细胞钙震荡特性

摘要

Objective:To ascertain the spontaneous and cyclical calcium oscillation in Cajal-like interstitial cells of Cajal in the bladder of adult guinea-pig.Method:The V-type enzyme collagenase was used to separate primary cells from myoblast of the preparation muscle tissue,primary cells were cultured.Using fluo-4 AM dye the Cells.The change of calcium fluorescence intensity was recorded with laser scanning microscope imaging. The changes of the intracellular calcium concentration was on behalf with the Fluorescence intensity.Result:The Cajal-like interstitial cells maintained its inherent characteristics by primary cultivation.We could see that the cell had a spindle cell body with two slender bipolars.In the Cajal-like interstitial cells of bladder:there were 2 different types of spontaneous calcium,small irregular calcium oscillation and large oscillation slow calcium oscillation.The second type of calcium oscillation was completely different from those of smooth muscle cells.Conclusion:We have confirmed the existence of the spontaneous calcium oscillation in the Cajal-like interstitial cells in the bladder of adult guinea-pig,that like the pacemaker cells of interstitial cells of Cajal in the digestive tract.Implying that part of the Cajal-like interstitial cells may be the pacemaker cells of contraction or activity of bladder.%目的：探讨正常成年豚鼠膀胱Cajal样间质细胞（Cajal-like interstitial cells）是否有自发周期性钙震荡（calcium oscillation），以期为膀胱兴奋性起搏细胞为Cajal样间质细胞这一学说提供实验依据。方法：运用胶原酶V消化法，获得原代豚鼠膀胱Cajal样间质细胞进行培养，采用激光共聚焦显微镜成像技术，Fluo-4AM负载后的Cajal样间质细胞内钙离子荧光强度的变化采用激光共聚焦显微镜成像技术进行记录，荧光强度的改变代表细胞内钙离子浓度的变化。结果：培养后的Cajal样间质细胞保持其固有特征，可见胞体为梭形且胞体两极有两个细长突起；培养的Cajal样间质细胞有两类不同的钙震荡：一类震幅小而频率不规则，另一类震幅大而频率慢。第二类钙震荡与膀胱逼尿肌细胞的钙震荡显著不同。结论：与消化道起搏细胞Cajal间质细胞（interstitial cells of Cajal，ICCs）自发性钙震荡类似，成年豚鼠膀胱Cajal样间质细胞亦存在自发钙震荡，提示膀胱收缩活动的起搏细胞可能为其中一种类型的膀胱Cajal样间质细胞。