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。     

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     

慢传输性便秘乙状结肠Cajal间质细胞分布的免疫组化观察

目的：Cajal间质细胞（interstitial cells of Cajal，ICC）对结肠动力有重要的调控作用。研究旨在初步观察ICC在STC乙状结肠的分布情况。方法：用ckit免疫组织化学技术观察了12例慢传输性便秘（slow transit constipation，STC）患者和8例对照组患者乙状结肠ICC形态和分布情况，并进行半定量分析。结果：ICC广泛分布于结肠肌层中，包括纵肌层（LM）、肌间丛（MP）、环肌层（CM）和黏膜下环肌表面（SMB）。ICC在形态上主要表现为双极细胞和多突起细胞两种类型。ICC—MP和ICC—CM相对多于ICC—SMB和ICC—LM。与对照组相比，STC患者各个区域ICC均明显减少（P〈0．01）。部分病例ICC—SMB几乎消失。结论：STC结肠ICC数量显著减少，但ICC减少是原发性的还是继发于某种其他损害仍有待探讨。     

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。     

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。     

大承气冲剂治疗对多器官功能不全综合征胃肠Cajal间质细胞形态学影响

目的:研究大承气冲剂在治疗多器官功能不全综合征(MODS)过程中对Cajal间质细胞(ICC)形态学变化的影响,探讨药物作用机制。方法:Wistar大鼠20只,随机分为MODS模型组和大承气冲剂治疗组。对各组大鼠取胃体、胃窦、小肠标本:(1)进行c-Kit免疫荧光染色,观察Cajal间质细胞在胃肠道的分布情况和数量变化;(2)进行电镜观察,明确ICC的超微结构变化。结果:治疗组与MODS组比较:胃肠肌间神经丛ICC的荧光染色分布较连续,保持条带样结构,细胞突起可见;细胞数量较MODS组增多。电镜下可见细胞核形态明显好于MODS组,存在大量线粒体,少量线粒体肿胀,细胞突起损伤不明显,与邻近平滑肌细胞、神经末梢和其他ICC之间的存在缝隙连接。结论:大承气冲剂可通过保护和维持ICC的正常形态和结构,维持细胞间信号正常传导,有效地改善胃肠运动障碍,增强胃肠蠕动。     

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间质细胞在大鼠"泻剂结肠"结肠肌电变化中的作用

目的　研究长期服用接触性泻剂对大鼠结肠肌电的影响 ,探讨Cajal间质细胞 (ICC)在“泻剂结肠”肌电变化中的作用。方法　 32只大鼠随机分为 2组 ,实验组饲以含酚酞饲料 ,3个月后测定结肠慢波频率及振幅 ;用碘化锌 锇酸法 (ZIO)观察肌间丛ICC变化 ,透射电镜观察肌间丛神经和ICC的超微结构变化。结果　“泻剂结肠”结肠慢波频率明显减慢 (P <0 .0 5 ) ,肌间丛ICC分布不均匀 ,突起连接杂乱 ;电镜下见肌间丛神经轴突空化 ,ICC样细胞变性。结论　长期服用酚酞可导致结肠慢波频率减慢 ,其可能机理为肌间丛神经及ICC变性所引起     

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.     

雌激素受体β在女性慢传输性便秘患者乙状结肠壁内的分布及其蛋白表达

目的研究女性慢传输性便秘(slow transit constipation,STC)患者雌激素受体β(estrogen receptor β,ERβ)在乙状结肠组织的分布及其蛋白表达。方法采用免疫组织化学及Western blot方法检测20例STC患者以及20例对照组患者乙状结肠ERβ的分布及其蛋白表达。结果免疫组化结果显示ERβ在STC组及对照组乙状结肠黏膜、肌间神经丛及黏膜下神经丛均有表达,肌层未见表达。与对照组相比,STC组ERβ表达均明显降低,差异有统计学意义(P0.05)。Western blot结果显示STC组乙状结肠ERβ蛋白表达较对照组明显降低,差异有统计学意义(P0.01)。结论肌间及黏膜下神经丛ERβ蛋白表达降低可能参与STC发病。     

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.     

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.     

Immunolocalization of the gap junction protein Connexin43 in the interstitial cells of Cajal in the normal and Hirschsprung's disease bowel

BACKGROUND: Interstitial cells of Cajal (ICC) are pacemaker cells between gastrointestinal smooth muscles; they generate spontaneous slow waves of the smooth muscle layers and mediate neurotransmission. The cellular network of ICC is connected by Gap junctions to each other and to the smooth muscle cells. Although there have been several studies reporting distribution of ICC in the normal bowel and pathological conditions such as Hirschsprung's disease, there is little information on the crucial role of Gap junctions in the intercellular communication in the gut musculature. The aim of this study was to investigate the immunolocalization of the Gap junction protein Connexin43 in the normal and Hirschsprung's disease (HD) bowel using whole-mount preparation technique and confocal laser scanning microscopy. METHODS: Full-thickness bowel specimens were collected at pull-through operation from 8 patients diagnosed as having HD. Normal control large bowel specimens were collected from 12 patients during bladder augmentation operation. Whole-mount preparation was performed on all specimens and double immunostaining was carried out using anti c-kit and antiConnexin43 antibodies. The immunolocalization was detected with the help of confocal laser scanning microscopy. RESULTS: Connexin43 immunoreactivity appeared in and between the c-kit-positive cells and along the smooth muscle fibers of the normal bowel and ganglionic part of HD bowel. In the aganglionic part of HD bowel there was no expression of Connexin43. In the transitional zone of HD the Connexin43 staining was weak and colocalized only in the processes of the c-kit-positive Cajal cells. CONCLUSIONS: Results of this study show for the first time that Gap junctional protein Connexin43 is present in the ICCs, which form a 3-dimensional network in the normal bowel wall. The lack of expression of Connexin43 in the aganglionic bowel and reduced expression in the transitional zone of HD suggest that the impaired intercellular communication between ICCs and smooth muscle cells may partly be responsible for the motility dysfunction in HD.     

Three-dimensional morphology of gut innervation in total intestinal aganglionosis using whole-mount preparation

BACKGROUND: Total intestinal aganglionosis (TIA) is a rare form of Hirschsprung's disease (HD). The aim of this study was to examine the 3-dimensional morphology of the myentric plexus of the entire gastrointestinal tract in a newborn with total intestinal aganglionosis. METHODS: Whole-mount preparations were made of the entire gastrointestinal tract using NADPH-diaphorase histochemistry and c-kit (a marker of interstitial cells of Cajal) immunohistochemistry. RESULTS: Whole-mount preparations of the esophagus, stomach, and duodenum showed 3-dimensional morphology of the myenteric plexus forming a meshlike network of nerve fibers, connected to each other and to ganglia. There were large numbers of NADPH-diaphrase-positive nerve fibers between the muscle fibers in the circular muscle layer. In esophagus, stomach, and duodenum, c-kit-positive interstitial cells of Cajal (ICC) formed a 3-dimensional network between the two muscle layers and also were abundant within the circular muscle layer. In the jejunum, ileum, and colon, the myenteric plexus was absent and was replaced by hypertrophic nerve bundles that stained weakly with NADPH-diaphrase. Circular muscle layer completely lacked NADPH-diaphrase-positive nerve fibers. The c-kit-positive ICCs in the jejunum, ileum, and colon were sparse and localized mainly around the nerve trunks between the circular and longitudinal muscle layers. CONCLUSIONS: Whole-mount preparation is an elegant 3-dimensional technique in which the relationship of branching and interconnecting nerve fibers to each other and to muscle can be seen clearly. Absence of myenteric plexus, lack of nitrergic innervation, and depletion of interstitial cells of Cajal in the bowel wall throughout the small and large bowel contribute to the inability of the smooth muscle to relax, thereby causing lack of peristalsis in TIA.     

How common is colonic elongation in children with slow-transit constipation or anorectal retention?

PurposeColonic elongation is reported as a possible cause for slow colonic transit, as it is observed in patients with slow-transit constipation (STC). This study aimed to determine the frequency of colonic elongation in children with STC or anorectal retention using radioimaging. We hypothesized that transverse colon elongation may occur in patients with STC, whereas sigmoid colon elongates in patients with anorectal retention.MethodsNuclear transit scintigraphy performed for chronic constipation (1999-2011) was analyzed qualitatively for elongated transverse colon or sigmoid colon. Three major colonic transit patterns were identified: slow transit in the proximal colon (STC), normal proximal colonic transit with anorectal retention (NT-AR), and rapid proximal transit ± anorectal retention (RT). χ² Test was used for statistical analysis (P < .05 significant).ResultsFrom 1999 to 2011, 626 children had nuclear transit scintigraphy. Transverse colon elongation occurred more frequently in STC (73/322, or 23%) compared with NT-AR (9/127, or 7%) and RT (5/177, or 3%; P < .0001). Sigmoid colon elongation was equally common in NT-AR (8/127, or 6%) compared with RT (10/177, or 6%) and STC (14/322, or 4%; P < .9).ConclusionTransverse colon elongation is more common in STC (23%), whereas sigmoid colon elongation is not more common in anorectal retention. Colonic elongation may be the cause or the result of the underlying slow colonic transit.     

Alterations in smooth muscle contractile and cytoskeleton proteins and interstitial cells of Cajal in megacystis microcolon intestinal hypoperistalsis syndrome

Background/Purpose: Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is characterized by decreased or absent peristalsis. Gastrointestinal motility depends on the enteric nervous system, smooth muscle cells (SMCs), and the interstitial cells of Cajal (ICCs). Contractile and cytoskeleton proteinase are important structural and functional components of SMCs. The aim of study was to examine the expression of contractile and cytoskeleton proteins in SMCs and distribution of ICCs in MMIHS bowel. Methods: Full-thickness bowel specimens were obtained from 4 infants with MMIHS and 4 controls. Specimens were processed as whole-mount preparations and frozen and paraffin sections. Combined staining of NADPH-d histochemistry/c-kit immunohistochemistry, single and double immunohistochemistry using [alpha ]-smooth muscle actin ([alpha ]-SMA), calponin (CALP), caldesmon (CALD), desmin (DES), protein gene product 9.5 (PGP 9.5) and c-kit antibodies were performed and examined using light and confocal scanning microscopy. Results: [alpha ]-SMA, CALP, CALD, and DES immunoreactivity were reduced markedly in MMIHS bowel compared with controls. Combined NADPH/c-kit staining showed dense network of ICCs around myenteric plexus in MMIHS bowel. In contrast, the intramuscular ICCs either were absent or reduced in MMIHS bowel. Conclusions: Marked reduction of contractile and cytoskeleton proteins in SMCs combined with reduced expression of intramuscular ICCs in the gut may be responsible for the motility dysfunction in MMIHS. J Pedriatr Surg 38:749-755. [copy ] 2003 Elsevier Inc. All rights reserved.