豚鼠膀胱Cajal样间质细胞的分离、培养及其鉴定

目的探索豚鼠膀胱Cajal样间质细胞的原代培养方法。方法颈部脱臼法处死豚鼠，无菌条件下分离出膀胱肌组织制备成肌条，采用V型胶原酶酶解法分离细胞，将细胞悬液接种于含干细胞因子的细胞培养基中培养，隔日换液。用酪氨酸蛋白激酶受体特异性抗体标记细胞，证实细胞类型。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，该类细胞酪氨酸蛋白激酶受体抗体荧光染色阳性证实细胞培养成功。结论用V型胶原酶消化法可获得成年豚鼠膀胱Cajal样间质细胞，并在体外条件下生长，可用于Cajal样间质细胞的电生理学研究。     

豚鼠膀胱组织内Cajal样间质细胞及NOS表达阳性神经元的分布

目的探讨Cajal样间质细胞和一氧化氮合酶2（NOS2）表达阳性神经元在成年豚鼠膀胱组织内的分布。方法成年豚鼠膀胱冰冻切片,c—Kit免疫荧光和n—NOS2免疫荧光染色。结果膀胱肌层内可见c—Kit表达阳性细胞的存在,n-NOS2免疫荧光染色显示其阳性神经元广泛分布于膀胱组织内。c—Kit免疫荧光和n-NOS2免疫荧光双重染色虽未发现两者的共存,但可见c—Kit表达阳性细胞存在于NOS表达阳性神经元的附近。结论豚鼠膀胱的Cajal样间质细胞可能参与膀胱自主节律性运动的调控,提示二者可能共同参与膀胱自主节律性运动的调控,为“神经-Cajal样问质细胞-肌肉”作用单元的形成提供结构基础。     

Cajal样间质细胞与糖尿病膀胱病变关系研究进展

糖尿病膀胱病变是因糖尿病而导致的排尿障碍,一般发病隐匿,症状进展极缓慢,容易被忽视.最近人们对它的认识在不断深入,发现膀胱Cajal样间质细胞在膀胱中起非常重要的作用,本文就Cajal样间质细胞在膀胱中主要特点、生理功能和SCF/c-kit信号通路对Cajal样间质细胞的影响,以及膀胱中Cajal样间质细胞异常在糖尿病膀胱病变发病机制的研究进展做一综述.     

Cajal样细胞在人类后腹腔镜下肾癌根治手术标本的分布特点

目的探讨Cajal样细胞在经后腹腔镜肾癌根治手术标本中的形态和分布特点。方法收集我科2008年1～8月行后腹腔镜肾癌根治手术切下的肾脏和输尿管上段标本23例,分别对肾盏、肾盂和输尿管上段取材、石蜡包埋、切片,常规HE染色及CD117免疫组织化学染色,以正常结肠肠管的Cajal细胞作为阳性对照组,光学显微镜下观察两者形态学的异同,以及Cajal样细胞在上尿路的分布密度特点。结果形态学上人类上尿路Cajal样细胞与结肠肌壁间的Cajal细胞相似,呈梭形,CD117染色阳性,不同之处在于结肠的Cajal细胞主要位于内环、外纵肌间的肌间神经丛周围,而人类上尿路Cajal样细胞散在分布于上尿路的固有层和肌层间。分布密度上,肾盏、肾盂和输尿管上段依次为15.4±5.4、22.6±6.6和19.9±5.8个/cm2,肾盏区Cajal样细胞的分布密度明显小于肾盂和输尿管上段（P=0.000,P=0.014）,而肾盂和输尿管上段Cajal样细胞的分布密度差异无显著性（P=0.129）。结论人类上尿路存在Cajal样细胞,但其分布范围与胃肠道Cajal细胞不同,在肾盏、肾盂、输尿管上段的分布密度也不相同,这种差异可能与其功能有关。     

豚鼠Oddi括约肌内Cajal样细胞及NOS表达阳性神经元的分布

目的研究Cajal样细胞和一氧化氮合酶(nitric oxide synthase,NOS)表达阳性神经元在成年豚鼠Oddi括约肌(sphincter of Oddi,SO)的分布。方法成年豚鼠SO冷冻切片,c-Kit免疫细胞化学和NADPH-黄递酶组织化学双重染色。结果SO横切面环行平滑肌层内可见少量c-Kit阳性细胞,胞体呈梭形,两端伸出细长的突起。纵切面SO的两个壁分别为:外侧壁和十二指肠壁内SO壁,前者与肠壁结构类似,在深肌丛和肌间神经丛周围可见较多的c-Kit阳性细胞;而十二指肠壁内SO壁的肌层内存在大量c-Kit阳性细胞和突起,其形态与肠壁肌层内Cajal细胞相似。NADPH-黄递酶组织化学染色可见NOS表达阳性神经元广泛分布于SO的肌间神经丛和平滑肌内。c-Kit免疫细胞化学和NADPH-黄递酶组织化学双重染色虽未发现二者的共存,但可见c-Kit阳性细胞及突起存在于NOS表达阳性神经元的附近。结论豚鼠SO内存在的Cajal样细胞可能参与SO自主节律性运动的调控,可能是NOS表达阳性神经元对SO运动发挥调节作用的靶细胞。     

尿路"Cajal样间质细胞"的研究进展

Cajal间质细胞(interstitial cells of Cajal, ICCs)具有独特和多变的形态学特征,是胃肠道慢波节律的起搏点.泌尿道平滑肌具有和胃肠道平滑肌相似的肌源性兴奋性,并且在泌尿系统多个部位陆续发现ICC样细胞,提示它们具有与胃肠道ICCs相似的功能.本文综述了泌尿道ICC样细胞的分布和特性以及它们与尿路动力性疾病的关系.     

人膀胱不同区域Cajal样间质细胞的分布及意义

目的观察人膀胱不同区域Cajal样间质细胞（interstitial cells of cajal,ICCs）的分布情况,探讨其作为起搏细胞在人膀胱的分布意义。方法标本来源于5个膀胱癌患者手术（全膀胱切除术）切下的正常非癌变膀胱全层组织（病理检查无病变）,按解剖学分组（顶部、体部、颈部）及组织学分组（黏膜层、黏膜下层、肌层）制作冰冻切片,运用免疫荧光显色技术观察ICCs的分布情况。结果通过激光共聚焦显微镜观察到在人膀胱不同区域发现的ICCs形态学、免疫表型与在消化道发现的ICCs类似。可见CD117呈阳性的ICCs出现在膀胱不同解剖区域及组织层次中,荧光主要在细胞膜及突起着色,细胞呈梭形,轴向两端存在突起。解剖学组中ICCs存在于膀胱顶部最多,膀胱体部次之,膀胱颈部罕见。组织学组中ICCs主要存在于肌层和黏膜下层,黏膜层少见。结论为临床上治疗某些疾病提供新的理论依据,ICCs在人膀胱顶部和肌层大量存在,很可能构成了膀胱活动的第一起搏点,使慢波向膀胱体、膀胱颈传播,缺乏及丧失ICCs可能会导致人类膀胱动力障碍性疾病。     

膀胱Cajal间质细胞研究近况

近年来,随着有关膀胱Cajal间质细胞(Interstitial cells of Cajal,ICCs)的不断深入研究,有关膀胱ICCs的分布、表面受体、信号传导、多种因素对其影响的多项研究证实:膀胱ICCs为膀胱的起搏细胞,且多种相关疾病:如糖尿病性膀胱病(DCP)、逼尿肌不稳定(DI)、膀胱出口梗阻(BOO)等均与膀胱ICCs异常有关,相关膀胱ICCs的研究为上述疾病的治疗提出新方向,现就膀胱Cajal间质细胞研究近况予以综述.     

膀胱Cajal间质细胞样细胞的研究进展

膀胱cajal间质细胞样细胞（interstitial cells of Cajal like cells,ICC-LC）是一种特殊间质细胞,它可能是膀胱逼尿肌的起搏者,其功能可能与某些膀胱功能障碍性疾病的发病有关,而成为尿动力方面研究的热点。本文就膀胱ICC-LC的分布、形态特征、电生理特征及与逼尿肌不稳定（detrusor instability,DI）的关系等方面国内外研究的新进展作一综述。     

膀胱间质细胞起搏机制的研究进展

膀胱间质细胞(IC)是区别于神经细胞和平滑肌细胞(SMC)的一类细胞。目前对IC的研究多聚焦于Cajal间质细胞(ICC)初始的起搏机制中,可认为IC中ICC表面存在产生超极化激活的内向电流的超极化激活环核苷酸门控阳离子通道,使得ICC成为膀胱起搏细胞成为可能。而成纤维样细胞和端粒细胞与膀胱平滑肌细胞和神经纤维之间存在...     

机械牵张对豚鼠膀胱组织Cajal间质细胞形态学及兴奋性的影响

目的 探讨机械牵张对豚鼠膀胱组织Cajal间质细胞(ICC)形态及兴奋性的影响.方法 建立雌性豚鼠膀胱颈部分梗阻(PBOO)模型作为实验组.并设假手术组作为对照.术后4周取膀胱组织制片,免疫荧光染色观察2组ICC形态和分布情况;胶原酶消化豚鼠膀胱制细胞悬液,免疫荧光标记,流式细胞仪检测2组c-kit阳性细胞比率.在弹性硅胶膜七原代培养膀胱ICC,利用Fluo-4钙荧光指示剂检测机械牵张对ICC钙信号的影响.结果 2组豚鼠膀胱组织铺片均可见c-kit染色阳性典型长梭形有突起的ICC,主要分布于平滑肌肌束间;PBOO组膀胱平滑肌间质增厚,c-kit染色阳性细胞数量及其突起明显增多,互相连接呈网络状;PBOO组c-kit阳性细胞比率为(6.7±1.7)%,显著高于对照组的(1.0±0.5)%,差异有统计学意义(P＜0.05).体外培养的ICC可检测到自发性钙波,机械牵张刺激可诱导ICC发生钙波增强现象.结论 机械牵张可诱导膀胱ICC兴奋性增强;PBOO膀胱组织中ICC数量及相互联系显著增多.ICC可能参与了膀胱牵张感受功能并在长期牵张应力负荷下发生一定的适应性改变.     

Interstitial cells of Cajal in the human normal urinary bladder and in the bladder of patients with megacystis-microcolon intestinal hypoperistalsis syndrome

OBJECTIVE: To investigate the distribution of c-kit-positive interstitial cells of Cajal (ICCs) in normal bladder and bladders from patients with megacystis-microcolon-intestinal peristalsis syndrome (MMIHS, a rare congenital and generally fatal cause of functional intestinal obstruction in the newborn), the most characteristic feature of which is abdominal distension caused by a distended unobstructed urinary bladder. PATIENTS AND METHODS: Full-thickness bladder specimens were obtained from four infants with MMIHS and four controls, and processed as paraffin-wax and frozen sections. Sections were assessed using single immunohistochemistry with monoclonal and polyclonal anti-c-kit antibodies. Anti-alpha-smooth muscle actin (SMA) antibody was used to investigate the contractile apparatus in smooth muscle cells of the urinary bladder. Specimens were examined using light and confocal scanning microscopy. RESULTS: There were many c-kit positive ICCs in the normal urinary bladder, appearing as small, long, bipolar cells with only two long and several short processes. In contrast, ICCs were absent in the MMIHS bladder. alpha-SMA immunoreactivity was lower in MMIHS urinary bladder than in control sections. CONCLUSION: This study shows for the first time the presence of c-kit-positive ICCs in the normal human urinary bladder. The lack of ICCs in the MMIHS bladder may contribute to the voiding dysfunction in this disease.     

二氨基乙氧基双苯萘酯对豚鼠膀胱不同区域逼尿肌兴奋性的影响

目的初步探讨二氨基乙氧基双苯萘酯（2-aminoethoxydiphenyl borate,2-APB）对豚鼠膀胱不同区域逼尿肌兴奋性的影响。方法将豚鼠膀胱按解剖标志划分并切取不同部位组织块,制成逼尿肌肌条,置Kreb液孵育,通过离体逼尿肌肌条实验观察2-APB对豚鼠逼尿肌收缩活动频率、幅度变化的影响。结果应用2-APB后膀胱不同区域逼尿肌收缩频率和收缩幅度抑制程度不同,顶部抑制程度最高,体部次之,颈部最弱。结论膀胱逼尿肌不同区域兴奋性不同,可能与不同区域ICCs样细胞的分布差异有关。     

Correlation between the structure and function of the rabbit urinary bladder following partial outlet obstruction

PURPOSE: To understand the relationship between contractile and structural changes in the obstructed bladder, rabbit bladder was partially obstructed for up to 70 days and alterations in tension response to field stimulation and carbachol were compared with alterations in ultrastructure and innervation of detrusor smooth muscle (SM). The effect of partial outlet obstruction on the physiological responses to field stimulation (FS) (nerve mediated contraction) and carbachol (receptor mediated contraction) were correlated with the structure and innervation of the detrusor smooth muscle (SM) of the same animal during a 70 day period. MATERIALS AND METHODS: 28 rabbits were subjected to 1 to 70 days of mild partial outlet obstruction. Sham operated rabbits were euthanized at 7, 14, 28, and 70 days post-obstruction. At each time period, isolated strips of bladder body were mounted in individual baths and the contractile response to FS and carbachol determined. Three additional strips from each bladder were fixed for electron microscopy. RESULTS: Bladder mass increased rapidly during the first 7 days after obstruction, was constant for the next 7 days, and then continued to increase gradually. Dysfunction of the contractile response to FS was noted as early as 3 days and progressively increased over the 70-day study period. The decrease in the response to FS increased at a significantly faster rate than the decrease in the contractile response to carbachol. In ultrastructure studies, at 3 and 7 days post-obstruction the majority of SM cells displayed the characteristics of hypertrophy. At 28 days some SM cells displayed loosely packed myofilaments and an irregular distribution of sarcoplasmic dense bodies. At 70 days swollen mitochondria were present in all cell types of the bladder wall. Evidence of axonal degeneration was first observed at 7 days post-obstruction and became more extensive thereafter. No evidence of mitotic figures, nerve growth cones or regenerating SM cells was observed. CONCLUSIONS: Prolonged partial bladder outflow obstruction is accompanied by a progressive decrease in contractility of SM. The present study describes the structural damage that occurs in the bladder wall in response to partial outlet obstruction and correlates these observations with the contractile dysfunction with which it is associated. Furthermore, mitochondrial damage in vessels and fibroblasts is suggestive of bladder wall ischemia.     

豚鼠膀胱cajal样细胞的类型

目的：观察豚鼠膀胱cajal样细胞形态学特点并结合该细胞具有的特殊功能,探讨cajal样细胞在膀胱组织中的类型。方法：豚鼠膀胱经手术取材后制成冷冻切片和体外培养膀胱cajal样细胞,采用间接免疫荧光染色后在激光共聚焦显微镜观察cajal样细胞形态学特点,然后将体外培养的膀胱cajal样细胞应用Fluo一4AM标记细胞内钙离子浓度含量,在激光共聚焦显微镜下观察cajal样细胞钙离子振荡功能。结果：膀胱组织冷冻切片和体外cajal样细胞培养均可见单体cajal样细胞和二聚体cajal样细胞,体外培养的两种类型的cajal样细胞具有钙离子振荡功能。结论：豚鼠膀胱组织中存在单体和二聚体两种不同类型的cajal样细胞。     

慢传输型便秘患者结肠壁内Cajal细胞的形态学研究

目的　慢传输型便秘 (STC)患者乙状结肠壁内Cajal细胞 (ICC)的形态学研究。方法　全层铺片、冰冻切片的免疫细胞化学染色及透射电镜观察。结果　正常成人乙状结肠壁内ICC主要分布在环肌内侧面与粘膜下层之间 (ICC SM)、环肌层内 (ICC CM)、纵肌层内 (ICC LM)及肌间神经丛周围 (ICC MP)。STC乙状结肠壁内ICC的数量均较正常对照组减少 ,其中 ,ICC SM和ICC CM减少尤为显著 ,约减少 6 0 %。铺片显示ICC -MP不仅数量减少 ,且突起的分支亦减少 ,彼此间不能形成完整的细胞网络。电镜观察可见上述部位的Cajal细胞内溶酶体聚集、脂质沉积 ,ICC SM突起间的缝隙连接较小、数量减少。结论　本研究结果提示ICC的这些病理改变可能与STC的发生、发展有关 ,但是 ,ICC的减少是该病的原因还是继发性损害的结果仍有待进一步研究     

Immunocolocalization of the heme oxygenase-2 and interstitial cells of Cajal in normal and aganglionic colon

Purpose: Interstitial cells of Cajal (ICCs) are pacemaker cells that play an important role in the control of gut motility. Carbon monoxide (CO) has been proposed as an endogenous messenger molecule between ICC and smooth muscle cells in the gastrointestinal tract (GIT). Heme oxygenase-2 (HO-2) is the main physiologic mechanism for generating CO in human cells. The aim of this study was to investigate the immunocolocalization of the HO-2 and ICCs in normal and aganglionic bowel of Hirschsprung's disease (HD). Methods: Full-thickness specimens were obtained from aganglionic colon during pull-through operation from 10 patients diagnosed as having HD. Normal control large bowel specimens were collected from 4 patients during bladder augmentation procedures. Double immunostaining was carried out using c-kit and HO-2 antibodies. Immunolocalization was detected by means of confocal laser scanning microscopy. Results: HO-2 immunoreactivity (IR) was found in many ICCs present around the myenteric plexus, within the longitudinal and circular muscle layers and at the innermost part of the circular muscle layer in normal colon. In the aganglionic colon there was absence of HO-2 IR in the sparsely found ICCs. In the transitional zone of HD bowel the colocalization of HO-2 IR and ICCs was much reduced compared with controls. Conclusions: The results of this study provide the first evidence for the presence of HO-2 immunoreactivity in the ICCs in normal human colon and absence of HO-2 immunoreactivity in sparsely appearing ICCs in the bowel of HD patients. The lack of HO-2 in the ICCs in the bowel of HD patients may result in impaired intracellular communication between ICCs and SMCs causing motility dysfunction. J Pediatr Surg 38:73-77.     

Abnormalities of C-Kit-positive cellular network in isolated hypoganglionosis

Background/Purpose: C-Kit-positive interstitial cells of Cajal (ICCs) have a key role in the normal motility function and development of the bowel. They are pacemaker cells, which facilitate active propagation of electrical events and neurotransmission in the bowel wall. ICCs are present in the bowel as myenteric ICCs (ICC_myS) and muscular ICCs (ICC_musS). The aim of this study was to examine the distribution of c-Kit-positive ICCs and their relationship to the autonomic intrinsic innervation in bowel specimens from patients with isolated hypoganglionosis. Methods: Full-thickness large bowel specimens were obtained from 6 patients with hypoganglionosis and from 4 patients during bladder augmentation (controls). Frozen sections and whole-mount preparations were stained using c-Kit immunohistochemistry, nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase, and acetylcholinesterase (AChE) histochemistry and evaluated using normal brightfield and confocal laser scanning microscopy. Results: NADPH-diaphorase and AChE histochemistry findings showed characteristic histologic features of hypoganglionosis, eg, sparse and small myenteric ganglia and low or absent AChE activity in the lamina propria. Myenteric plexus in the normal bowel was surrounded by a dense network of c-Kit-positive ICC_myS, whereas in hypoganglionosis sparse isolated ICC_myS were found. C-Kit-positive ICC_musS were reduced markedly in the longitudinal and circular muscle layer and at the innermost part of the circular muscle in hypoganglionosis. Conclusion: Deficient expression of c-Kit-positive myenteric and muscular ICCs in the hypoganglionic colon may contribute to the motility dysfunction in the affected bowel.