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     

慢传输性便秘患者乙状结肠组织中Cajal间质细胞的分布

目的　了解Cajal间质细胞 (ICC)在慢传输性便秘 (STC)患者乙状结肠组织中的分布。方法　应用c kit单克隆抗体间接免疫荧光检测技术 ,对 12例STC患者和 8例对照组患者的乙状结肠组织中的ICC分布进行测定 ,激光共聚焦显微镜扫描图像 ,图像处理软件进行分析。结果　ICC广泛分布于结肠肌层中 ,包括纵肌层、肌间丛、环肌层和黏膜下环肌表面 ,形态主要表现为双极细胞和多突起细胞两种类型 ;肌间丛和环肌层ICC数量多于黏膜下环肌表面和纵肌层。与对照组相比 ,STC患者各个区域ICC均明显减少 (P <0 0 1)。部分病例黏膜下环肌表面ICC几乎消失。结论　STC患者结肠存在ICC减少 ,但ICC减少是原发性还是继发性有待探讨     

Cajal间质细胞在先天性巨结肠分布的研究

目的：通过观察cajal间质细胞（interstitial cells of cajal，ICC）在正常结肠及先天性巨结肠先天性巨结肠（hirschsprung’s disease，HD）患者痉挛段、移行段、扩张段的分布，探讨HD的发病机制。方法：收集25例HD患儿标本，于术中分别选取扩张段、移行段、痉挛段肠壁的全层组织，另取6例手术患儿的正常结肠全层组织标本，常规固定石蜡包埋组织切片备用。对标本行c-Kit免疫组织化学染色。光镜观察ICC的分布，计数并进行统计学分析。结果：正常结肠ICC主要分布在环肌内侧面与黏膜下层之间即黏膜下ICC（submucosal ICC．ICC—SM）、环肌与纵肌之间的肌间神经从周围即肌间ICC（myenteric ICC，ICC—MY）以及环肌与纵肌内。HD患儿痉挛段ICC—SM、ICC—IM细胞数较扩张段和正常对照组明显减少（P〈0．01），且ICC的细胞突起的分支亦减少，彼此之间不能形成完整的细胞网络。而扩张段ICC与正常对照组比较无明显差异（P〉0．05）。结论：HD患儿结肠ICC的异常分布，可能是HD发病、肠管蠕动障碍以及排便异常的原因之一。     

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60％。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。     

自制复方黄芪汤治疗慢传输型便秘与C—kit结肠组织表达的研究

为研究结肠慢传输型便秘（STC）大鼠结肠组织Cajal间质细胞（ICC）的分布、形态和数量变化,以及与大鼠结肠组织中CkitmRNA的表达的关系,观察自制汤剂复方黄芪汤治疗STC模型大鼠的疗效,将大鼠32只随机分为两组：模型组（24只）和生理盐水对照组（8只）,模型组饲料中添加大黄粉制作便秘模型,生理盐水对照组饲以普通软饲料。停药一周后从模型组中随机选取8只,与生理盐水对照组同时禁食不禁水24h后,测活性炭推进率,处死,取出标本。测模型组结肠组织中ICC的数量和形态改变,以及组织中C—kitmRNA的表达改变,并与生理盐水对照组进行比较。另外16只随机分为生理盐水空白组（8只）和中药干预组（8只）,中药干预组灌服自制汤剂复方黄芪汤10ml／（kg·d）,约含生药2g／ml。灌药30d。生理盐水空白组灌服同体积的生理盐水30d,测活性炭推进率,处死,取出标本。测中药干预组结肠组织中ICC的数量和形态改变以及组织中C—kitmRNA的表达改变,并与生理盐水空白组进行比较。结果显示,与生理盐水对照组比较。模型组炭末推进长度小于生理盐水对照组（P〈O．05）,炭末推进率明显低于生理盐水对照组（P〈O．01）。Cajal间质细胞的形态变短、变钝,细胞数量变少（P〈O．05）。Ckit mRNA的表达量变少。与生理盐水空白组比较,中药干预组炭末推进长度大于生理盐水空白组（P〈0．05）,炭末推进率明显高于生理盐水空白组（P〈0．01）。ICC形态恢复正常,细胞数量增多（P〈0．05）。C—kitmRNA的表达量变多（P〈0．05）。结果表明,大黄饲喂法造模是成功的,STC大鼠结肠组织中存在ICC的形态变短、变钝,细胞数量变少,CkitmRNA的表达量变小。自制汤剂复方黄芪汤能使STC大鼠结肠组织中ICC形态和数量向正常转化,使C—kitmR—NA的表达量增多。     

Cajal间质细胞在先天性巨结肠和巨结肠同源病结肠中的分布研究

目的研究先天性巨结肠（Hirschsprung’s disease，HD）和巨结肠同源病（allied Hirschsprung’s disease，AHD）肠壁内Cajal间质细胞（interstitial cells of Cajal，ICCs）的分布状态，探讨HD和AHD的发病机制。方法选择确诊为HD和AHD的患者各20例，取巨结肠根治术吻合口远端的全层肠壁作为实验组，另取16例正常结肠标本作为对照组。用鼠抗人c—kit单克隆抗体（CD117）标记ICCs，Image Pro-Plus图像分析系统检测ICCs。结果对照组中大量ICCs分布在肌间神经丛周围和环纵肌层内，ICCs包绕神经丛周围，肌层间ICCs连续分布；HD组远端肠管中肌层间和各肌层内ICCs明显减少甚至缺如，与对照组比较差异有统计学意义，P〈0．01；AHD组远端肠管中神经丛大小不一，ICCs分布差异大，大多数神经丛区ICCs减少，环肌层内ICCs明显减少，与AHD组和对照组比较差异有统计学意义，P〈0.01。HD组远端结肠中ICCs减少比AHD组显著，差异有统计学意义，P〈0．01。结论HD、AHD病变肠管中除了神经节细胞的异常外，同时存在ICCs异常；ICCs在HD和AHD的分布不同可能与两者临床症状差异有关；肠管中ICCs数量可能与临床症状及预后有一定关系。     

慢传输性便秘结肠平滑肌肌动蛋白改变

目的：深入地了解慢传输性便秘的发病机理和病理生理改变。方法：以免疫组化方法检测了STC患者结肠平滑肌收缩蛋白－肌动蛋白的变化。结果：与正常对照组相比,慢传输性便秘患者结肠黏膜下肌层、纵肌层内α肌动蛋白量明显减少（P＜0．01）,而环肌层内α肌动蛋白变化不明显（P＞0．05）。结论：推测这种平滑肌收缩蛋白的减少和分布异常导致了慢传输性便秘病人结肠运动功能的紊乱。     

结肠慢传输型便秘的研究现状及展望(摘要)

结肠慢传输型便秘(STC)是临床上常见的、以腹胀及便意淡漠为主要症状的慢性顽固性便秘。近年来大量的临床和实验研究发现。(1)STC病人的结肠壁变薄、肌细胞空泡变性或脂肪变性、环肌萎缩,病变呈进行性过程。(2)肠壁问神经节细胞数量减少,排列紊乱,形态皱缩或轻度水肿,空泡变性;神经微丝和微管数量减少、排列紊乱。(3)肠壁内兴奋性神经递质(Ach、SP)减少,抑制性递质(VIP)含量有增高现象。(4)Cajal间质细胞(ICC)的分布和功能异常与肠动力障碍有密切关系,详细机理尚不清楚。根据研究资料,肠壁肌细胞和肌问神经丛的损害、神经递质的改变以及ICC的分布和功能异常是STC发病的关键环节,究竟什么原因引起这些病理改变,目前尚不清楚。因此,目的STC的预防尚无良策,治疗仍是一般性的保守治疗,包括:①粗纤维饮食;②在结肠高动力期(早晨起床后或早餐后)训练排便运动,改善排便体位(蹲位最佳);③加强腹肌和膈肌锻炼;①适当给予粪便软化剂和润肠剂;⑤上述治疗无效者给予低渗性药物、水灌肠或油剂保留灌肠。对保守治疗无效者给予手术治疗,目的理想的术式为全结肠切除,回一直肠吻合术和次伞结肠切除,盲一直肠吻合术。作者提出。在STC确诊后先用结肠壁活组织病理检查或电生理等检查。根据肠壁肌、肌间神经、ICC及神经递质的     

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。     

大承气汤对MODS大鼠小肠深部肌间Cajal间质细胞损伤的作用

目的：观察多器官功能障碍综合征（MODS）大鼠小肠深部肌间Cajal间质细胞（ICC—DMP）的形态学变化，探讨大承气汤治疗MODS的机制。方法：Wistar大鼠50只，随机分为对照组、MODS模型组和大承气汤治疗组。采用免疫荧光标记、激光扫描共聚焦显微镜结合透射电镜观察MODS大鼠小肠ICC—DMP立体网络结构和超微结构的变化以及大承气汤治疗后的改变，并对ICC—DMP分布进行定量分析。结果：MODS组小肠ICC—DMP网络结构连接不完整，Cajal间质细胞超微结构损伤明显。大承气汤治疗组小肠ICC—DMP网络结构较MODS组完整，ICC—DMP超微结构无明显损伤。结论：大承气汤能够通过修复MODS大鼠小肠ICC—DMP形态学的损伤，改善MODS大鼠胃肠运动障碍。     

Assessing interstitial cells of Cajal in slow transit constipation using CD117 is a useful diagnostic test

Slow transit constipation (STC) is a colonic motility disorder that is characterized by measurably delayed movement of stools through the colon. The pathophysiology of STC is unclear and both the interstitial cells of Cajal (ICC) and cells of the enteric nervous system are believed to play an important role. The aim of this study was to compare the number and distribution of ICC and cells of the enteric nervous system in patients with a control group by means of immunohistochemistry. Formalin-fixed paraffin-embedded colonic sections were obtained from 15 patients, aged between 23 and 52 (mean age=37 y), who underwent colectomy for STC. Forty-five cases of normal colon from age and sex-matched nonobstructive colorectal cancer patients were selected as controls. By using c-kit (CD117) and PGP 9.5 immunohistochemical studies, ICC and enteric neurofilaments were demonstrated, respectively. The number of cells were counted under 40 x high-power field (HPF) in 3 layers of the colonic muscularis propria, that is, the inner circular muscle layer, the myenteric plexus, and the outer longitudinal muscle layer in both test and control groups. The mean number of ICC and enteric neurofilaments were significantly reduced in all 3 layers of the muscularis propria from STC patients compared with controls. This reduction was most significant in the inner circular muscle layer (P<0.0001). A cutoff value of 7 ICC per HPF in the inner circular muscle layer can be used as a further confirmation to the clinical diagnosis of STC in resected specimens.     

豚鼠阴茎海绵体中ICC和NO信号通路的形态学关系

目的初步探讨豚鼠阴茎海绵体组织中cajal间质细胞（ICC）和神经型一氧化氮合成酶（nNOS）阳性神经元的分布及两者的组织学关系。方法①制作成年豚鼠阴茎海绵体组织冰冻切片,利用c-kit/nNOS免疫荧光双染技术,激光共聚焦扫描显微镜（LSCM）下观察阴茎海绵体ICC及nNOS阳性神经元的形态及分布情况。②酶消化法体外培养阴茎海绵体ICC,c-kit/nNOS免疫荧光双染,LSCM下观察细胞形态及c-kit和nNOS的表达情况。结果 ICC分布在阴茎海绵体平滑肌小梁边缘及平滑肌肌间,纺锤形,具有长的突起,胞体呈圆形或椭圆形,并且表达nNOS;阴茎海绵体平滑肌小梁边缘及平滑肌肌间同时分布有ICC和nNOS阳性神经元,局部nNOS阳性神经主干附近可见较多ICC分布。结论豚鼠阴茎海绵体ICC与nNOS阳性神经元组织联系紧密,ICC可能参与阴茎海绵体内NO神经信号的传递。     

大鼠小肠Cajal间质细胞的体外分离、培养及鉴定

目的：探讨大鼠小肠Cajal间质细胞（ICC）的原代分离、培养及鉴定方法。 方法：出生后5~10 d的SD乳大鼠,颈椎脱臼处死。无菌条件下取出小肠,在解剖显微镜下剥去小肠系膜、肠黏膜和黏膜下层,将小肠平滑肌层组织剪成小块后接种于含有干细胞因子的DMEM培养基中进行培养。倒置显微镜下连续观察组织块周围细胞的游出情况及细胞的形态,用酪氨酸蛋白激酶受体c-kit特异性抗体免疫荧光染色鉴定细胞类型。 结果：培养1周后,倒置显微镜下可见组织块周围长出细胞,呈梭形、三角形,有多个短突起;随着培养时间的延长,突起细长化并彼此相互连接形成网络。该类细胞c-kit抗体免疫荧光染色呈阳性。 结论：该研究成功建立了一套由大鼠小肠平滑肌组织块原代培养ICC的方法,为ICC的生物学特性及其与胃肠道动力障碍性疾病关系的研究奠定了基础。     

便秘二号方对慢传输型便秘大鼠Cajal间质细胞及C-KIT表达的影响

目的:观察便秘二号方对结肠慢传输型便秘大鼠模型的疗效,并检测其对大鼠模型结肠组织Cajal间质细胞及C-KIT表达的影响。方法:32只大鼠随机分为正常组、模型组、实验组、盐水对照组,每组8只。正常组给予正常饮食,其余3组采用大黄递增灌胃法制作慢传输型便秘大鼠模型。造模成功后,实验组予中药便秘二号方(方由当归、白术、黄芪、升麻、杏仁、枳实、肉苁蓉组成)灌胃,盐水对照组予生理盐水灌胃。30d后以活性炭灌胃法检测结肠传输功能,以免疫组化法观察结肠组织Cajal间质细胞的形态和数量,采用RT-PCR法观察C-KITmRNA表达。结果:炭末推进长度模型组明显少于正常组(P0.05),实验组明显高于盐水对照组(P0.05),推进率比较差异均有统计学意义(P0.01);实验组与盐水对照组相比,可见各肌层Cajal间质细胞数量增加,细胞形态恢复;实验组与盐水对照组的Cajal间质细胞个数比较,实验组高于对照组,差异有统计学意义(P0.01);模型组C-KIT表达阳性率明显低于正常组,实验组C-KIT表达阳性率明显高于盐水对照组。结论:中药便秘二号方可以增强慢传输型便秘大鼠的结肠蠕动功能,增加结肠组织中Cajal间质细胞的数量,改善其形态,促进C-KIT表达。     

Origin, course, and endings of abnormal enteric nerve fibres in Hirschsprung's disease defined by whole-mount immunohistochemistry

Accurate delineation of the intramural pathway of abnormal enteric nerve fibres in Hirschsprung's disease has previously proved impossible because the neural network is invariably transected in conventional histological sections. With the technique of wholemount immunohistochemistry (WI), the bowel segment is converted into a rectangular sheet and the serosa, long muscle (LM), circular muscle (CM), submucosa, and mucosa are separated into layers to allow each nerve plexus to be examined intact and neural pathways traced. The entire resected bowel specimens of nine HD infants and five infants serving as controls were investigated, using neuron-specific enolase and vasoactive intestinal peptide (VIP) for WI. The major new findings are (1) More VIP fibres were observed in aganglionic bowel with WI than with conventional sections; (2) Thick nerve trunks in aganglionic bowel do not descend from intrinsic neurons of oligoganglionic bowel as previously suggested, but have an extrinsic origin, accompanying blood vessels as small nerves initially, expanding subsequently, and ending blindly in submucosa; (3) CM nerve fibres follow muscle fibres concentrically for long distances in aganglionic bowel; and (4) LM nerve fibres meander in spirals in aganglionic bowel instead of running straight. This study shows that (1) WI is highly sensitive; (2) nerve fibres in aganglionic bowel have an extrinsic origin; and (3) innervation abnormalities in Hirschsprung's disease are not only quantitative but qualitative.     

Kit positive cells in the guinea pig bladder

PURPOSE: We describe the presence of interstitial cells of Cajal (ICC) throughout the wall of the guinea pig bladder. MATERIALS AND METHODS: Bladders obtained from male guinea pigs were prepared for immunohistochemical investigations using various primary antibodies, including the specific ICC marker c-kit (Gibco BRL, Grand Island, New York). Enzymatically dispersed cells with a branched morphology were identified as ICC using anti-c-kit. They were loaded with fluo-4acetoxymethyl (Molecular Probes, Eugene, Oregon) and studied using confocal laser scanning microscopy. RESULTS: Anti-c-kit labeling demonstrated that ICC were oriented in parallel with the smooth muscle bundles that run diagonally throughout the bladder. Double labeling with anti-smooth muscle myosin (Sigma Chemical Co., St. Louis, Missouri) revealed that ICC were located on the boundary of smooth muscle bundles. When anti-c-kit was used in combination with the general neuronal antibody protein gene product 9.5 (Ultraclone Ltd., Isle of Wight, United Kingdom) or anti-neuronal nitric oxide synthase, it was noted that there was a close association between nerves and ICC. Enzymatic dissociation of cells from tissue pieces yielded a heterogeneous population of cells containing typical spindle-shaped smooth muscle cells and branched cells resembling ICC from other preparations. The latter could be identified immunohistochemically as ICC using anti-c-kit, whereas the majority of spindle-shaped cells were not Kit positive. Branched cells responded to the application of carbachol by firing Ca2+ waves and they were often spontaneously active. CONCLUSIONS: ICC are located on the boundary of smooth muscle bundles in the guinea pig bladder. They fire Ca2+ waves in response to cholinergic stimulation and can be spontaneously active, suggesting that they could act as pacemakers or intermediaries in the transmission of nerve signals to smooth muscle cells.     

Loss of CD117 (c-kit)- and CD34-positive ICC and associated CD34-positive fibroblasts defines a subpopulation of chronic intestinal pseudo-obstruction

Chronic idiopathic intestinal pseudo-obstruction is a syndrome in which symptoms of intestinal obstruction are present in the absence of mechanical obstruction. Lack of normal pacemaker activity, usually generated by the interstitial cells of Cajal (ICC), could account for the apparent obstruction. ICC are normally located around and between the myenteric plexus ganglia and within muscle and also in the deep muscular plexus of the small bowel and the submuscular plexus of the large intestine, just within the circular muscle. ICC can be demonstrated immunohistochemically with CD117 (c-kit) as well as with CD34, although this is less specific. CD34 also stains a population of fibroblasts that are intimately associated with ICC. To determine whether there is a relative deficiency of ICC and CD34-positive fibroblasts in patients with chronic idiopathic intestinal pseudo-obstruction, tissue from 30 patients of large intestine and eight patients with small intestine pseudo-obstruction was obtained. Controls (large intestinal specimens from 12 patients, small intestinal specimens from six patients) were chosen from resections for Crohn's disease and colorectal neoplasia, both with and without dilatation. Examination of pseudo-obstruction cases identified 10 patients (nine large intestinal and one small intestinal) in which both CD117 and CD34 were absent or severely reduced in all three of the examined areas. In contrast, the control cases, including those with preobstructive dilatation, showed relatively constant ICC staining. These results suggest that there is a proportion of pseudo-obstruction cases in which the ICC are markedly reduced. These results also demonstrate that, in these cases, loss of the kit immunoreactivity is correlated with the loss of CD34 staining: this indicates that both the ICC and the CD34-positive fibroblasts associated with the ICC are absent. These findings will allow surgical pathologists to identify this subpopulation of patients with CIIP using tissue obtained by laparoscopic biopsy of the muscularis propria or surgical resection.     

Cajal间质细胞在大鼠慢传输便秘模型结肠中的变化

目的研究Cajal间质细胞（ICC）与慢传输型便秘（STC）的关系。方法用复方地芬诺酯灌胃的方法建立大鼠慢传输便秘模型（STC组）,Westernblot法测定STC组与对照组大鼠升结肠和降结肠组织ICC的特异性标志物c—kit变化情况,利用其与对应的内参B—actin灰度值的比值作为各组c—kit蛋白相对含量。结果STC组大鼠日均粪便量为（1．3±0．7）g／100g,比对照组的（1．6±0．9）g／100g明显减少,差异有统计学意义（t=10．798,P〈0．05）。STC组首粒黑粪排出时间为（461．6±150．8）min,较对照组大鼠的（351．3±119．9）min显著延长（t=2．291,P〈0．05）。STC组与对照组升结肠c—kit灰度比平均值分别为0．277±0．077和0．576±0．081（t=10．719,P〈0．05）;降结肠c—kit灰度比平均值分别为0．280±0．075和0．571±0．079（t=10．700,P〈0．05）;c．kit在STC组升结肠和降结肠的表达均下调,差异均有统计学意义。结论ICC在升结肠和降结肠的减少可能对慢传输型便秘的发生与发展有一定作用。