【作者】 朱国栋；

【导师】 王勤章；

【作者基本信息】 石河子大学 ， 外科学， 2009， 硕士

【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。更多还原

【Abstract】 Purpose:The purpose of this is to provide proofs to prove that the Cajal-like cells in guinea pig bladder is the pacemaker by observe its utrastructure and electrophysiological, and offer a new starting point linical and treatment of certain diseases (such as the detrusor instability, DI).Methods:In this experimental we can get the cajal-like interstitial cells by enzyme digestion method. We observe ICCs ultrastructure by transmission electron microscope; Cells were loaded with fluo-4 AM dye. Using laser scanning microscope imaging records the change of calcium fluorescence intensity. Fluorescence intensity was on behalf of changes of the intracellular calcium concentration; by patch clamp technology We observe the calcium current of ICCs in the whole-cell recording mode. It can provide a proof to verify that the ICCs in guinea pig bladder is the pacemaker.Results:In this experimental the ICCs can be observed to have rich in mitochondria, developed the RER, the sliding surface endoplasmic reticulum, Gorky body. There were 2 different types of spontaneous calcium oscillation in the Cajal-like interstitial cells of bladder:large, slow calcium oscillation and small, irregular calcium oscillation. The first type of calcium oscillation was completely different from those of smooth muscle cells; By the patch clamp technology We can’t observe the type calcium current of ICCs in the whole-cell recording mode.Conclusion:The ICCs in guinea pig bladder have a unique ultrastructure, which similar with the ICCs’s in the gastrointestinal tract. We have confirmed that part of the Cajal-like interstitial cells existence of the spontaneous calcium oscillation in the bladder of adult guinea-pig, which like the pacemaker cells of interstitial cells of Cajal in the digestive tract. The spontaneous calcium oscillation was not observed in urinary bladder detrusor. Implying that part of the.Cajal-like interstitial cells may be the pacemaker cells of contraction or activity of bladder. The role of the other part of the Cajal-like interstitial cells in urinary bladder possibly plays the conduction, the coupling information.更多还原