大鼠胃肠道不同部位Cajal间质细胞与慢传输型便秘的关系

目的观察慢传输型便秘(slow transit constipation,STC)大鼠模型胃肠道Cajal间质细胞(interstitial cell of cajal,ICC)的分布特点与含量改变,全面评估ICC在STC发病机制中的作用。方法 24只健康Wistar大鼠随机分成便秘组和对照组,分别饲喂含复方苯乙哌啶的混悬液和生理盐水,每5 d记录1次大鼠大便粒数、大便干质量及大鼠体质量。饲养90 d后停药1周,测定胃肠道传输功能并通过免疫组化的方法测定ICC的特异性标志物c-kit+细胞在胃窦、小肠、结肠的分布情况与含量变化。结果便秘组日均粪便粒数少于对照组(P<0.01),平均每粒粪便质量大于对照组(P<0.05);便秘组首粒黑便排出时间长于对照组(P<0.05);与对照组比较,便秘组胃窦部位c-kit+细胞数量无明显变化(P>0.05)。而c-kit+细胞在便秘组大鼠小肠、结肠的数目均少于对照组(P<0.05)。结论在STC模型中,胃窦ICC变化不明显,小肠ICC数量有减少趋势,可能对STC有一定影响,结肠部位ICC数量明显减少,可能是慢传输型便秘大鼠的主要病理生理机制。     

Cajal间质细胞与慢传输型便秘研究的现状与趋势

Cajal间质细胞(ICC)是分布在胃肠道神经细胞与平滑肌之间的一种特殊细胞,具有起搏及协调胃肠道运动的功能。研究发现慢传输型便秘(STC)患者结肠组织中Cajal间质细胞数量减少,形态及其网状结构改变,因此ICC的异常可能是STC的重要发病机制。预防或干预ICC的病变,积极改善ICC的功能,协调肠道运动可能是潜在治疗STC的方向。     

糖尿病慢传输运动结肠Cajal间质细胞和干细胞因子的变化

目的:了解Cajal间质细胞(ICC)、干细胞因子(SCF)在糖尿病大鼠结肠慢传输运动模型中的变化,探讨其作用及可能的调控机制.方法:54只♂ SD大鼠分为糖尿病和正常对照组,经腹腔注射链脲佐菌素建立糖尿病大鼠模型,于造模后6,8,10 wk各组分别处死9只大鼠,以免疫组化、透射电镜研究近端结肠组织中ICC的变化,以Western blot方法检测近端结肠组织中膜结合型干细胞因子的表达,以ELISA测定血清中可溶型SCF的浓度,分析他们之间的相关性.结果:糖尿病大鼠血糖随时间增加而升高,而胃肠推进率却降低(P＞0.05).免疫组化结果显示,6,8,10 wk时的糖尿病大鼠肌间ICC表达较对照组明显减少(P＜0.05),且糖尿病大鼠近端结肠ICC数量随时间推移有逐渐降少的趋势.透射电镜显示糖尿病大鼠结肠ICC线粒体肿胀、空泡样变,细胞器数量明显减少.与对照组相比,糖尿病大鼠血清中可溶型SCF显著降低(6 wk:0.93±0.53 μg/L vs 1.87±0.92 μg/L,P＜0.05;8 wk: 0.78±0.21 μg/L vs 1.76±0.94 μg/L,P＜0.05;10 wk:0.73±0.20 μg/L vs1.82±0.96 μg/L,P＜0.05),而结肠组织中的膜结合型SCF无明显差异(P＞0.05),且可溶型干细胞因子与ICC具有相同的变化趋势.结论:糖尿病胃肠动力障碍大鼠存在血清中可溶性干细胞因子浓度下降以及结肠组织中ICC数量减少和结构破坏,这些变化及其可能存在的序贯性调控作用可能是糖尿病出现结肠慢传输变化的基础.     

Slow transit colon constipation is not related to the number of interstitial cells of Cajal

Background and aims Recent studies have demonstrated decreased numbers of interstitial cells of Cajal in patients suffering from severe chronic constipation as measured by c-Kit (CD117) and CD34 immunohistology. In this study, we wished to determine whether there were abnormalities in the number of neurons of the Auerbach's plexus, their CD117 and CD34 immunoreactivity, or the thickness of colon wall sections in patients with refractory slow transit colonic constipation as compared with control subjects.Patients and methods Specimens from 13 patients who had undergone subtotal colectomy for severe chronic constipation refractory to medical treatment were compared with normal controls. Enteric neurons of Auerbach's plexus were counted, and thickness of the circular and longitudinal layer of the muscularis externa as well as total muscularis externa was measured. Quantitative assessment of anti-CD117 and anti-CD34 immunoreactivity was performed using an Automated Cellular Imaging System and expressed as fractional scores.Results Except for a decreased circular muscle layer thickness in the constipated patients, no statistically significant differences were observed between the two groups. In particular, there was no relationship between CD117/CD34 fractional staining score and the duration or severity of disease, despite the selection of highly symptomatic individuals requiring colonic resection.Conclusion Using quantitative immunohistochemistry for CD117/CD34, we could not detect a relationship between fractional CD117/CD34 staining score and chronic constipation as compared to controls.     

小鼠不全性肠梗阻对Cajal间质细胞表型的影响

目的:探讨小鼠小肠慢性不全性肠梗阻导致自发性节律性收缩运动、电活动变化与Cajal间质细胞(ICC)表型转化的关系.方法:通过外科手术方法在小鼠回肠套入硅胶管,建立小鼠不全性肠梗阻模型.利用苏木精-伊红(H&E)染色,研究小肠平滑肌层形态学改变;利用常规生理和电生理技术观察环行平滑肌肌条自发性节律性收缩和慢波变化规律;利用全层平滑肌免疫组织化学方法,观察ICC表型标志酪氨酸激酶受体(c-kit)表达的变化.结果:小鼠不全性肠梗阻模型形成后14d,梗阻近端肠管显著扩张、小肠平滑肌层明显增生肥厚;小肠平滑肌自发性节律性收缩节律紊乱、幅度不规则、频率变慢;平滑肌静息膜电位去极化、慢波幅度变小、频率变慢;ICC表型标志c-kit表达显著减弱、甚至消失.结论:小鼠不全性肠梗阻导致机械和电活动紊乱与ICC表型转化导致的ICC数量减少和网络破坏有关.该模型为进一步研究ICC表型改变的细胞/分子机理提供了一个良好的实验模型.     

Decreased density of interstitial cells of Cajal and neuronal cells in patients with slow-transit constipation and acquired megacolon

BACKGROUND: The pathophysiology of constipation is not clearly identified as yet, and the interstital cells of Cajal (ICC), known to generate the slow wave activity and to be involved in intestinal neurotransmission and the enteric nervous system (ENS), are suspected to play an important role. The aims of the present study were to assess the distribution of ICC and neuronal cells of ENS in patients with slow-transit constipation and acquired megacolon. METHODS: Sigmoid colon specimens were obtained from patients who underwent colectomy due to slow-transit constipation (n = 10), acquired megacolon (n = 9) and non-obstructive colon cancer (n = 10) as a control group. The ICC were visualized by c-Kit immunohistochemistry and neuronal cells of the ENS were demonstrated by protein gene product (PGP) 9.5. Density of cells stained by c-Kit and PGP 9.5 was calculated as percent area (area stained/area of X-Y plane) x 100, when images were collected at a magnification of x40 objective, with maximum area examined in the horizontal X-Y plane of 400 microm x 400 microm using an image analyzer. RESULTS: The densities of ICC and PGP 9.5 reactive neuronal structures were significantly decreased in all layers of sigmoid colon specimens in patients with slow-transit constipation and acquired megacolon, compared with that of the control group. However, there was no statistically significant difference in either the density of ICC or that of neuronal structures between the patients with slow-transit constipation and acquired megacolon. CONCLUSIONS: Slow-transit constipation and acquired megacolon were associated with alteration of ICC and neuronal cells of ENS in the sigmoid colon.     

慢传输型便秘患者结肠中五羟色胺受体亚型的表达及意义

目的:研究主要5-HT受体亚型在慢传输型便秘(slowtransitconstipation,STC)患者结肠中的表达,探讨其在慢传输型便秘发病机制中的作用.方法:采用免疫组化EnVision法,检测20例STC患者和20例对照组结肠组织中5-HT1A,5-HT3和5-HT4受体的分布及表达水平,并采用IMS计算机辅助图像分析系统进行半定量分析.结果:5-H1A受体分布于黏膜下层、肌层,肌间神经丛5-HT1A受体的表达在STC组和对照组间无显著差异(P=0.548).5-HT3受体分布于黏膜下层和肌层,肌间神经丛STC组阳性指数显著低于对照组(65.2±15.9vs94.3±20.1,P<0.01).5-HT4受体分布于黏膜层、黏膜下层、肌层.在黏膜层和肌间神经丛,STC组5-HT4受体阳性指数均显著低于对照组(57.8±10.9vs78.5±12.9,P<0.01;77.5±19.9,119.2±26.9,P<0.01).STC组中,5-HT3受体表达水平与结肠传输试验第5天体内残留标志物数量无关(P>0.05);但5-HT4受体表达水平与第5天体内残留标志物数量呈负相关(r=-0.782,P<0.01).结论:STC患者结肠中存在5-HT3和5-HT4受体亚型的表达下调,两者可能参与了STC的发病机制.     

Disruption of interstitial cells of Cajal networks after massive small bowel resection

AIM: To investigate the disruptions of interstitial cells of Cajal (ICC) in the remaining bowel in rats after massive small bowel resection (mSBR). METHODS: Thirty male Sprague-Dawley rats fitting entry criteria were divided randomly into three experimental groups (n = 10 each): Group A rats underwent bowel transection and re-anastomosis (sham) and tissue samples were harvested at day 7 post-surgery. Group B and C rats underwent 80% small bowel resection with tissue harvested from Group B rats at day 7 post-surgery, and from Group C rats at day 14 postsurgery. The distribution of ICC at the site of the resid-ual small bowel was evaluated by immunohistochemical analysis of small intestine samples. The ultrastructural changes of ICC in the remnant ileum of model rats 7 and 14 d after mSBR were analyzed by transmission electron microscopy. Intracellular recordings of slow wave oscillations were used to evaluate electrical pacemaking. The protein expression of c-kit, ICC phenotypic markers, and membrane-bound stem cell factor (mSCF) in intestinal smooth muscle of each group were detected by Western blotting. RESULTS: After mSBR, immunohistochemical analysis indicated that the number of c-kit-positive cells was dramatically decreased in Group B rats compared with sham tissues. Significant ultrastructural changes in ICC with associated smooth muscle hypertrophy were also observed. Disordered spontaneous rhythmic contractions with reduced amplitude (8.5 ± 1.4 mV vs 24.8 ± 1.3 mV, P = 0.037) and increased slow wave frequency (39.5 ± 2.1 cycles/min vs 33.0 ± 1.3 cycles/min, P = 0.044) were found in the residual intestinal smooth muscle 7 d post mSBR. The contractile function and electrical activity of intestinal circular smooth muscle returned to normal levels at 14 d post mSBR (amplitude, 14.9 ± 1.6 mV vs 24.8 ± 1.3 mV; frequency, 30.7 ± 1.7 cycles/min vs 33.0 ± 1.3 cycles/min). The expression of Mscf and c-kit protein was decreased at 7 d (P = 0.026), but gradually returned to normal levels at 14 d. The ICC and     

慢传输型便秘结肠次全切除术后盲肠直肠吻合方式的探讨

目的探讨慢传输型便秘行结肠次全切除术后更合理的盲肠直肠吻合方式。 方法对兰州军区兰州总医院连续收治并进行手术治疗的34例STC患者的临床资料进行回顾性分析,男性3例,女性31例;年龄36～75岁,平均60.5岁。所有患者均选择行结肠次全切除盲肠直肠吻合,手术在常规开腹或是腹腔镜辅助下进行,盲肠直肠吻合方式采用回盲瓣对侧的盲肠侧壁与直肠残端进行端侧吻合。 结果传统开腹施行手术18例,腹腔镜辅助手术16例。术后1～3天开始排便,6～20次/d,术后第5-6天时排便次数逐渐减少。22例患者在术后10天时大便次数减少至8次/d以内;10例患者术后出现稀水样便伴肛门疼痛,经口服易蒙停、思密达等对症处理后好转。术后发生吻合口瘘1例、切口感染5例、尿潴留2例、炎性肠梗阻2例。30例患者得到随访2月至8年,大便次数保持在1-6次/d。2例患者7年后诉肛门坠胀,排便不净感。1例患者于术后4.5年时出现不完全性肠梗阻,经保守治疗后痊愈。27例患者术后立即感觉治疗效果满意,占80%;随访过程中,所有患者自觉症状均较术前改善明显,对手术治疗满意。无围手术期死亡。 结论对于确实需要手术治疗的STC患者,结肠次全切除盲直端侧吻合可以作为术式选择之一。     

肠神经系统与慢传输型便秘

慢传输型便秘(STC)病因未明,多因素与其发病相关.肠神经系统(ENS)可独立调节肠道功能,其在慢传输型便秘中的改变具有重要意义.此文就此予以阐述,为STC临床治疗提供依据.     

Abnormalities of nerve fibers in the circular muscle of patients with slow transit constipation

Abnormalities of the enteric nervous system are thought to explain the pathophysiology of motility disorders. Our aim was to determine if particular classes of enteric neurons are affected in slow transit constipation (STC). Specimens were taken from the terminal ileum and ascending, transverse and descending colon of patients undergoing subtotal colectomy for STC. Immunohistochemistry was performed using antisera to neuron-specific enolase, tachykinin, leu-enkephalin, choline acetyltransferase, vasoactive intestinal peptide, nitric oxide synthase, tyrosine hydroxylase and neuropeptide Y. The density of nerve fibres labelled with these antibodies in each layer was compared with age-matched controls. The density of nerve fibres with tachykinin and enkephalin immunoreactivity was reduced in the colonic circular muscle of the 15 patients with STC, whereas innervation of all other layers was normal. This reduction of tachykinin-immunoreactive nerve fibres also occurred in nine of the 12 specimens of terminal ileum examined. No difference was detected in the density or distribution of nerve fibres using the other antisera. Excitatory nerve fibres are present in the circular muscle in STC but they are deficient in tachykinins and enkephalin. Accepted: 14 January 1998     

糖尿病大鼠胃肠功能紊乱时结肠组织中Cajal间质细胞的表达

目的观察糖尿病大鼠胃肠功能紊乱时结肠组织内Cajal间质细胞的分布及表达变化,探讨Cajal间质细胞在糖尿病胃肠功能紊乱发病机制中的作用。方法30只SD大鼠随机分为两组,糖尿病组20只,正常对照组10只。糖尿病组大鼠用链脲佐菌素单剂量腹腔注射建立糖尿病模型,对照组注射等量枸橼酸缓冲液。两组大鼠饲养6周后处死,计算胃肠推进率并且收集结肠组织标本。用免疫组化方法观察Cajal间质细胞在两组大鼠结肠组织内的分布和表达,用W estern b lot方法检测c-k it蛋白在两组大鼠结肠内的表达。结果糖尿病组大鼠胃肠推进率较对照组明显降低（P〈0.05）。免疫组化和W estern b lot检测都显示糖尿病大鼠结肠组织内Cajal间质细胞的表达较正常大鼠明显减少（P均〈0.05）。结论糖尿病大鼠结肠组织内Cajal间质细胞表达减少,推测与糖尿病大鼠胃肠功能紊乱有一定相关性。     

针刺治疗老年慢传输型便秘60例临床观察

目的 观察针刺治疗老年慢传输型便秘的疗效及对肠动力的影响.方法 随机将60例老年慢传输型便秘患者分为两组,其中,实验组：30例老年患者运用针刺治疗,每日1次,每次30分钟.对照组：30例给予口服莫沙必利治疗,每次5mg,每日三次.两组均共治疗2周.结果 实验组30例,痊愈14例（46.67％）,有效15例（50％）,无效1例（3.33％）,总有效率96.67％;对照组30例,痊愈8例（26.67％）,有效16例（53.33％）,无效6例（20％）,总有效率80％,实验组疗效优于对照组,两组差异有统计学意义（P＜0.05）.结论 针刺治疗老年慢传输型便秘具有较好的临床疗效,并能有效促进肠蠕动.     

艾灸联合穴位注射对慢传输型便秘患者胃动素的影响

目的:研究艾灸联合穴位注射治疗慢传输型便秘的临床疗效及其对慢传输型便秘患者胃动素(MLT)的影响,探讨其治疗慢传输型便秘的可能机制.方法:将符合纳入标准的56例慢传输型便秘患者随机分为观察组和西药组.同时选取正常体检志愿者28人作为正常对照组.西药组患者给予莫沙比利10 mg口服,3次/d,共20 d.观察组患者给予双...     

小鼠空肠cajal间质细胞的分离与培养

目的研究CD1小鼠空肠cajal间质细胞(ICC)的分离和培养方法,为进一步研究其生理学特征提供基础。方法无菌条件下取小鼠空肠组织,分离出平滑肌肌条,采取组织块培养法,在24孔培养板中进行原代培养,光镜下观察其形态,用荧光标记的特异性C_Kit抗体进行鉴定。结果倒置显微镜下观察ICC有多个突起,并有次级分支,相互之间形成网络。免疫荧光法可见ICC呈阳性。结论组织块培养法可以简便、有效地培养ICC。所培养的ICC细胞生物学特性有待进一步研究。     

Hand-assisted laparoscopic total colectomy for slow transit constipation

Purpose Current medical treatments for slow transit constipation (STC) are often ineffective, and total colectomy with ileorectal anastomosis has been the procedure of choice for selected patients with refractory STC. Today, minimally invasive approaches are being utilized in a greater number of procedures as surgeons become more familiar with the techniques involved. The aim of this study was to assess the safety and utility of hand-assisted laparoscopic total colectomy for STC. Method From January 2002 to December 2005, 44 women presented with complaints of intractable constipation and failed to respond to medical treatment. Slow transit constipation was diagnosed after a series of examinations, including a colonic transit test, anal manometry, balloon expulsion test, and barium enema. All eligible patients underwent a hand-assisted laparoscopic total colectomy with ileorectal anastomosis. Main outcome measures included the operative time, conversion to open procedure, blood loss, time to return of flatus, length of postoperative hospital stay, and complications. Result The mean operative time was 197 min (range, 125–295 min). The mean estimated blood loss was 113 ml (range, 100–300 ml). The mean day of first time to flatus was 2 days, and the mean hospital stay was 7.6 days. There was no conversion to an open procedure and no surgical mortality. In the following period, two patients developed intestinal obstruction, which underwent exploratory laparotomy. However, some 39 patients (88.6%) expressed excellent or good in satisfaction. Conclusion Hand-assisted laparoscopic total colectomy could be a safe and efficient technique in the treatment of STC.     

慢传输型便秘大鼠结肠诱导型一氧化氮合酶和血红素氧合酶2的变化

目的 探讨慢传输型便秘(STC)大鼠结肠内诱导型一氧化氮合酶(iNOS)和血红素氧合酶2(HO-2)的变化.方法 健康Wistar大鼠32只,随机分为STC组和对照组,每组16只.采用复方苯乙哌啶灌胃法制备STC大鼠模型,饲养100 d后,采用活性炭灌胃法测定肠道传输速度确定模型建立.用免疫组织化学方法分别检测iNOS和HO-2在大鼠结肠的表达情况.结果 STC组大鼠日均粪便粒数、日均粪便干重、日均粪便质量均比对照组明显减少;检测大鼠肠道传输速度,STC组较对照组明显减慢,首粒黑便排出时间较对照组显著延长.iNOS和HO-2在STC大鼠结肠的表达明显强于对照组.结论 iNOS和HO-2在STC大鼠结肠的表达异常,表明iNOS和HO-2在慢传输型便秘发病机制中可能起着重要的作用.     

Cajal间质细胞的研究进展

Cajal间质细胞(interstitial cells of Cajal,ICC)是一类主要分布于胃肠道的间质细胞,是胃肠道的起搏细胞(pacemaker cell)和信号传导细胞,与肌细胞以及末梢神经元有着紧密的关系,具有激发和促进胃肠蠕动的作用.借助于电子显微镜技术,清楚观察到了ICC位置分布和内部精细结构;应用免疫荧光等生化技术,发现了其特殊表达的C-kit蛋白;利用电生理技术,得知多种胃肠动力障碍疾病也与其异常有关.多年来,学者逐渐在胃肠道、胆道、膀胱、子宫等部位发现了ICC的踪迹,并试图阐述其与某些疾病的发生机制.本文就ICC的起源、形态学、受体和功能、以及与其相关的疾病等作一综述.