豚鼠膀胱Cajal样细胞超微结构及起搏功能的相关性研究

发布时间：2018-01-14 13:17

本文关键词：豚鼠膀胱Cajal样细胞超微结构及起搏功能的相关性研究　出处：《石河子大学》2009年硕士论文　论文类型：学位论文

【摘要】：目的：本课题以正常成年豚鼠膀胱Cajal样间质细胞为研究对象,通过对膀胱Cajal样间质细胞(interstitial cells of Cajal, ICCs)超微结构观察及电生理学特性研究,探讨其是否为膀胱的起博细胞,为临床上治疗某些疾病(如detrusor instability, DI)提供新的切入点。方法:本实验运用采用酶消化法,得到体外培养状态下的ICCs样细胞,透射电镜观察正常成年豚鼠膀胱ICCs样细胞的超微结构；采用激光共聚焦显微镜成像技术记录Fluo4AM负载后的Cajal样间质细胞内钙离子荧光强度的变化,以荧光强度代表细胞内钙离子浓度的变化。；运用膜片钳技术,在全细胞记录模式下观察其是否具有钙电流,以验证其是否为膀胱的起博细胞。结果：透射电镜观察到CCs样细胞胞浆内线粒体、粗面内质网较丰富,滑面内质网分散分布,中间丝难能见到,未见微管；运用激光共聚焦显微镜成像技术连续扫描,见体外培养的原代膀胱Cajal样间质细胞有2种类型钙震荡：一类振幅大、频率慢而规则的钙震荡；另一类振幅小、频率快且不规则,且形成网络的膀胱Cajal样间质细胞的钙离子震荡的幅度明显高于单个的膀胱Cajal样间质细胞。膀胱逼尿肌细胞钙震荡振幅小不规则,其振幅显著低于Cajal样间质细胞第一类钙震荡振幅；膜片钳试验膀胱Cajal样细胞的钙离子电流不明显。结论：电镜观察到Cajal样细胞具有独特的超微结构,与胃肠道中ICCs的超微结构有相似；部分膀胱Cajal样细胞具有自发而规律的钙震荡,类似消化道起搏细胞Cajal间质细胞(interstitial cells of Cajal,ICCs)的自发性钙震荡,膀胱逼尿肌无自发性钙震荡。提示：部分膀胱Cajal样细胞是膀胱收缩活动的起搏细胞；另外部分膀胱Cajal样细胞可能起到传导、耦合信息的作用。
[Abstract]:Objective: to study the Cajal like stromal cells of normal adult guinea pig bladder. The ultrastructure and electrophysiological characteristics of interstitial cells of Cajal (ICCs) were observed. To investigate whether it is a primordial cell of the bladder and to provide a new entry point for the clinical treatment of certain diseases such as detrusor instability. Methods: ICCs like cells were obtained by enzyme digestion in vitro. The ultrastructure of ICCs like cells in normal adult guinea pig bladder was observed by transmission electron microscope. Laser confocal microscopy (LSCM) was used to record the changes of intracellular calcium fluorescence intensity in Cajal like stromal cells after Fluo4AM loading, and the fluorescence intensity represented the change of intracellular calcium concentration. The patch-clamp technique was used to observe whether it had calcium current in whole cell recording mode to verify whether it was a pacemaker cell of the bladder. Results: the mitochondria in the cytoplasm of CCs like cells were observed by transmission electron microscope, the rough endoplasmic reticulum was abundant, the smooth endoplasmic reticulum was scattered, the middle filaments could not be seen, and no microtubules were found. Continuous scanning with laser confocal microscopy showed that there were two types of calcium oscillations in primary bladder Cajal like stromal cells: one was large amplitude, slow frequency and regular calcium oscillation; The other has small amplitude, fast frequency and irregular frequency. The amplitude of calcium oscillation in the mesenchymal cells of bladder Cajal was significantly higher than that in the single interstitial cells of Cajal. The amplitude of calcium oscillation in the bladder detrusor muscle cells was small and irregular. The amplitude of calcium oscillation was significantly lower than that of Cajal like stromal cells. Patch clamp test showed that calcium current in Cajal-like cells of bladder was not obvious. Conclusion: the ultrastructure of Cajal like cells is similar to that of ICCs in gastrointestinal tract. Some Cajal like cells of bladder have spontaneous and regular calcium concussion. Spontaneous calcium concussion similar to interstitial cells of Cajal interstitial cells in digestive tract pacemaker cells. There was no spontaneous calcium concussion in the detrusor of the bladder. It was suggested that some of the bladder Cajal like cells were pacemaker cells of bladder contraction. Another part of bladder Cajal-like cells may play a role in the transmission and coupling of information.
【学位授予单位】：石河子大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R334

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