摘要
目的探讨外源性ATP对大鼠膀胱Cajal间质细胞(ICC)兴奋性的影响。方法采用胶原酶消化法急性分离SD大鼠膀胱ICC和逼尿肌细胞(DSMC),通过激光共聚焦技术检测外源性ATP作用前后ICC及DSMC内Ca2+信号的变化。结果胶原酶P消化后,在干细胞因子(SCF)的刺激下,经c-k it免疫荧光染色阳性的ICC呈长梭形或纺锤形,胞体较小,并且带有分枝状突起,而DSMC呈纤维形或短棒状,二者形态有明显区别。F luo-4 NW染色后,激光共聚焦显微镜观察显示,静止状态下ICC内[Ca2+]i平均荧光强度(55.16±3.23)显著强于平滑肌细胞(45.59±2.08),P<0.05;20μmol/L和100μmol/L ATP可诱发体外培养的ICC及DSMC细胞内[Ca2+]i一过性升高,随后缓慢下降。结论在SCF刺激条件下,SD大鼠膀胱ICC和DSMC可在体外稳定混合培养。外源性ATP可兴奋体外混合培养的ICC和DSMC。
Objective To explore the effect of ATP on excitability of interstitial cells of Cajal（ICC） and detrusor smooth muscle cells（DSMC） in SD rat bladder.Methods ICC and DSMC in SD rat bladder were isolated by collagenase digestion and the effects of ATP on variation of Ca2＋ in ICC and DSMC were assessed by confocal laser scanning microscopy（CLSM）.Results Obvious morphological differences were observed between ICC and DSMC.The positive ICC were long shuttle-shaped or spindle-shaped,small with branch-like protrusion while those of DSMC were in fiber shape or short-bar shape after immunoflurorescence staining by c-kit.The results of CLSM showed that the mean intensity of Ca2＋ in ICC was（55.16±3.23）,obvious higher than that of（45.59±2.08） in DSMC under static condition after Fluo-4 NW staining（P〈0.05）.Continue application of ATP at concentrations of 20 μmol/L and 100 μmol/L caused a transient rise of Ca2＋followed by a slow decline in both ICC and DSMC.Conclusion ICC and DSMC in SD rat bladder can be co-cultured in vitro under stem cell factor（SCF） stimulation and ATP can excite ICC and DSMC in vitro.
出处
《局解手术学杂志》
2011年第2期117-119,共3页
Journal of Regional Anatomy and Operative Surgery
基金
国家自然科学基金资助项目(30700270
81070604)
作者简介
[作者简介]樊汝会（1968-），女，重庆市人，主管技师，本科，主要从事排尿功能障碍方面的研究。电话：（023）68765346
[通讯作者]方强，电话：（023）-68765345，E-mai：fangqiang@tmmu．edu．cn
