胆道Cajal间质细胞的研究进展

Cajal间质细胞(interstitial cells of cajal,ICC)作为首先在胃肠道内发现的一类特殊的间质细胞,近几年在胃肠道外,尤其胆道系统内也普遍发现ICC的存在.通过复习文献,本文就其在胆道系统的ICC的一般生物学特性、尤其在胆道分布的特殊性,以及与胆道疾病的关系等方面的研究进展作一综述.     

阻断c-kit对小鼠小肠Cajal间质细胞的影响及机制初探

目的:观察阻断c-kit对小鼠小肠Cajal间质细胞(ICC)的影响以及与Cajal间质细胞c-kit基因表达的关系。方法:选取新出生Balb/c小鼠,腹腔内隔天注射c-kit抗体(ACK2)100μg,共5次,对照组注射生理盐水,于小鼠出生后第10天观察小肠Cajal间质细胞的分布情况并检测小肠肌条c-kitmRNA及蛋白的表达。结果:阻断c-kit后小鼠小肠中未能明显观察到ICC分布,并且c-kitmRNA表达以及c-kit蛋白表达明显减少。结论:阻断c-kit可导致小鼠小肠Cajal间质细胞缺失,与ICCc-kit基因水平表达下调引起ICC发育受影响有关。     

慢性综合应激对大鼠肠道Cajal间质细胞的影响

目的 观察慢性综合应激对大鼠行为及Cajal间质细胞(ICC)的影响.方法 将雄性Wistar大鼠30只随机分为对照组(15只)和模型组(15只).模型组接受21d不同的应激,对照组不予任何应激.造模前后行旷场试验.整个实验期间,定期观察大鼠体质量及行为变化.采用透射电镜和免疫荧光法观察各组大鼠ICC超微结构及网络变化.结果 模型组大鼠体质量(230.50±14.89)g明显低于对照组(292.50±20.55)g(P＜0.05);旷场试验显示,模型组大鼠总行程(1407.68±586.92)cm和直立次数(7.13±2.50)较正常对照组(6224.58±1918.25)cm、(17.07±5.42)明显减少,差异有统计学意义(P＜0.05).慢性综合应激使大鼠ICC超微结构发生改变、数量减少、与周围其他ICC、平滑肌和神经末梢之间的缝隙连接显著减少.结论 大鼠ICC形态结构改变,可能是慢性综合应激所致功能性胃肠病而引起胃肠运动异常的重要环节.

Abstract：

Objective To investigate the effect of chronic and comprehensive stress on the behavior and interstitial cells of Cajal (ICC) of adult rats. Methods Thirty male Wistar rats were randomly divided into 2 groups: normal control group (n = 15) and model group (n = 15). Except the normal group,the model group was given various kinds of stimulation for continuous 21 days. Open-field test was used before and after the chronic and comprehensive stress. During the whole experiment, the body weight and behavior of each rat were observed in periodic days. The ultrastructure and network of ICC in adult rats were observed by using the transmission electron microscopy and immunofluorescence. Results Compared with the control group, the body weight in the model group was remarkably reduced [ ( 230. 50 ± 14. 89 ) vs (292.50 ± 20. 55 ) g, P ＜ 0. 05 ) ]. Before and after chronic stress, there were significant differences in total ambulation distance and total number of rearings in open-field tests [ (6224.58 ± 1918. 25 ) vs ( 1407.68 ± 586. 92) cm, ( 17.07 ± 5.42) vs (7. 13 ± 2. 50), P ＜ 0. 05 ) ]. The ultrastructure of ICC was changed, and the quantity of ICC and gap junctions between ICC and surrounding cells, such as other ICC,smoth muscle cells, and terminal nerves were decreased significantly in the model group as compared with the contro group. Conclusion The changes of ICC may play an important role in abnormal gastrointestinal motility of functional gastrointestinal diseases caused by chronic and comprehensive stress.     

肝硬化大鼠小肠Cajal间质细胞超微结构及平滑肌细胞线粒体膜电位的变化

目的研究肝硬化大鼠小肠Cajal间质细胞超微结构的改变及平滑肌细胞线粒体膜电位的变化,探讨肝硬化胃肠动力障碍的相关机制。方法 20只Wister大鼠随机分为肝硬化模型组和对照组,每组10只,采用CCl4溶剂大腿根部皮下注射法制作肝硬化模型,用葡聚糖蓝-2000作为胃肠内标记物,观察大鼠肠道传输,透射电镜观察小肠Cajal间质细胞超微结构的改变,激光共聚焦显微镜检测小肠平滑肌细胞线粒体膜电位的改变。结果与对照组比较,肝硬化组大鼠小肠动力明显减弱(P0.01),透射电镜观察显示空肠Cajal间质细胞的超微结构发生明显改变,线粒体等细胞器明显减少(P0.01)。小肠平滑肌线粒体膜电位显著低于对照组(P0.01)。结论肝硬化大鼠肠道传输减弱与肠道Cajal间质细胞超微结构改变,细胞器减少及线粒体膜电位的降低有关。     

大黄素对体外培养Cajal间质细胞钙离子振荡功能的影响

目的:探讨大黄素对体外培养Cajal间质细胞钙离子振荡的影响,从而进一步揭示大黄素等促进胃肠动力的分子机制方法:体外培养小鼠ICC,体外添加大黄素,应用钙离子指示剂标记细胞内钙离子含量,在激光共聚焦显微镜下动态观察其钙离子振荡功能的变化。结果:免疫荧光鉴定结果证实体外培养细胞为ICC,体外培养ICC具有钙离子振荡功能,大黄素能显著增强其钙离子振荡的频率和振幅。结论:大黄素能显著增强体外培养ICC钙离子振荡功能,可能是其促进胃肠动力的重要机制。     

Cajal间质细胞的分离、培养方法探讨

目的　探讨体外培养Cajal间质细胞 (interstitialcellsofCajal,ICC)的方法。 方法　以小鼠小肠为ICC的组织来源 ,应用酶解法并结合密度离心法分离、培养ICC ,对ICC培养中的污染、Ca2 、Mg2 离子以及胶原酶等影响因素进行了探讨 ,应用ICC特异的c kit抗体进行免疫荧光染色后在激光共聚焦显微镜对其进行了鉴定。结果　成功建立起一套的分离、培养的方法。免疫荧光鉴定结果提示 ,本方法建立的ICC分离培养方法是稳定可靠的。结论　ICC分离培养比较困难 ,需要注意防止污染、减少无Ca2 、Mg2 离子液体以及胶原酶消化时间等问题 ,本方法有望为致力于ICC细胞研究的科研工作者提供方法借鉴 ,并有望促进ICC研究的深入进行。     

豚鼠胆囊胆固醇结石形成过程中胆囊Cajal样间质细胞及动力的变化

目的 观察豚鼠胆囊胆固醇结石形成过程中胆囊组织Cajal样间质细胞(ICLCs)及胆囊动力的变化.方法 将100只豚鼠随机分为实验组与对照组,每组50只,实验组给予致石饲料(胆固醇含量2％),对照组给予正常颗粒饲料,8周造模结束后,利用胆囊全层铺片免疫荧光化学染色及激光共聚焦显微镜观察各组ICLCs数量的变化,利用透射电镜观察各组胆囊组织中ICLCs的超微结构特征,利用在体胆囊收缩素(CCK-8)激发实验评价胆囊动力的变化.结果 对照组豚鼠胆囊ICLCs平均阳性面积为(18.37±0.64)％,实验组为(2.47±0.28)％,较对照组明显降低(t=21.122,P＜0.01);对照组豚鼠胆囊胆汁时间-流量曲线下面积为(31.58 ±2.26)μl,实验组为(15.02±1.98)μl,较对照组明显降低(t=8.586,P＜0.01).结论 饮食诱导的豚鼠胆囊胆固醇结石形成过程中,胆囊ICLCs明显减少,同时胆囊动力受损,对外源性CCK反应下降.胆囊动力受损可能与ICLCs减少有关.     

电针足三里穴对不全肠梗阻大鼠小肠Cajal间质细胞影响的免疫组化观察

目的初步观察电针足三里穴对不全肠梗阻大鼠小肠Cajal间质细胞(interstitial cells of Cajal,ICC)数量变化的影响,为进一步探讨电针足三里穴对ICC表型变化的影响奠定基础。方法采用圈套法造成不全肠梗阻从而建立ICC数目减少的SD大鼠模型。取20只雌性大鼠采用简单随机法均分成正常对照组、不全肠梗阻30 d未电针足三里穴组(梗阻组)、不全肠梗组30 d电针足三里穴组(足三里组)和不全肠梗阻30 d电针阴陵泉穴组(阴陵泉组)4组。其中足三里组和阴陵泉组分别连续电针足三里穴或阴陵泉穴7 d后,取小肠组织采用免疫组化方法以及免疫荧光观察ICC数量的变化。结果正常对照组、足三里组、梗阻组及阴陵泉组ICC阳性面积分别为(102 051.00±16 969.38)μm2、(92 642.12±14 854.49)μm2、(45 221.33±6 230.20)μm2和(63 136.16±8 863.91)μm2,各组间差异有统计学意义(F=21.240,P<0.001),其中足三里组的ICC阳性面积较梗阻组高(P<0.05)。结论电针足三里穴可使不全肠梗阻大鼠小肠受损的ICC数量得到部分恢复,但其具体机理有待进一步研究。     

硬膜外输注吗啡联合小剂量纳洛酮对兔结肠Cajal间质细胞的影响

目的 探讨硬膜外输注吗啡复合小剂量纳洛酮对兔肠蠕动及结肠组织中Cajal间质细胞密度变化的影响.方法 健康家兔60只,采用随机数字表法将其分为四组:即生理盐水对照组(NS组)、吗啡对照组(M组)、吗啡复合纳洛酮组(NM组)和纳洛酮组(N组),每组15只.四组均行硬膜外穿刺置管,背硬膜外镇痛泵,连续输注7d.观察记录粪便性状以及疼痛评分.测量幽门至回盲部肠管长度及墨汁在肠道的推进距离,计算墨汁推进率.取近端和远端结肠组织,采用免疫组化法测定c-kit表达,行HE染色,光镜下观察病理学结果,进行Cajal间质细胞(ICC)计数.结果 与M组比较,N组、NS组疼痛评分明显升高,小肠推进率明显升高,近端结肠c-kit表达和ICC计数明显增多(P＜0.05).与NM组比较,N组、NS组疼痛评分明显降低(P＜0.05);M组疼痛评分明显升高,小肠推进率明显降低,近端结肠c-kit表达和ICC计数明显降低(P＜0.05).近端结肠组织M组出现ICC损伤细胞数明显高于其他三组,c-kit阳性细胞面积明显小于其他三组.结论 硬膜外输注吗啡镇痛可抑制家兔肠运动功能,其机制与降低近端结肠ICC数量有关;小剂量纳洛酮可能逆转吗啡诱导的ICC变化,增强镇痛效果并减少其不良反应的发生.     

Cajal间质细胞与慢传输型便秘的关系

Cajal间质细胞(interstitial cells of cajal,ICC)是主要分布在消化道自主神经末梢与平滑肌细胞之间的一类特殊细胞,是胃肠运动的起搏细胞,具有产生、传导电慢波,调节胃肠道平滑肌运动的功能.深入研究Cajal间质细胞,对进一步认识胃肠运动的生理及胃肠动力疾病的发生机制有重要意义.     

胆囊癌合并肝门胆管侵犯的手术治疗体会

目的探讨胆囊癌合并肝门胆管侵犯手术治疗需注意的问题.方法回顾性分析22例手术治疗病人的术前发现和手术情况,比较切除与未切除受累胆管组术后生存时间.结果手术证实全组均有明确胆管侵犯.未切除胆管者病情无实质性改善,切除胆管组术后生存时间较长(P＜0.01),但仍存在"扩大的姑息性切除”的情况;胰后淋巴结清扫不彻底是妨碍实现临床根治的最主要原因.结论对胆囊癌合并肝门胆管侵犯者可根据有无肝门横沟处肝实质浸润、左右肝管是否显像、门静脉主干及左右支有无受累决定是否施行扩大根治切除;对适合手术切除的晚期胆囊癌宜将淋巴清扫扩大至第3站,以求达到真正意义上的临床根治.     

The expression of Cajal cells at the obstruction site of congenital pelviureteric junction obstruction and quantitative image analysis

Objective

The study aimed to analyze the expression and significance of interstitial cells of Cajal (ICC) at the obstruction site of congenital pelviureteric junction obstruction (PUJO).

Methods

Specimens of the strictured segment of the PUJ were obtained from 24 patients who were diagnosed to have PUJO (without aberrant vessels and adhesive band compression of ureteral junction) intraoperatively. In the control group, PUJ specimens were taken from 21 patients who had Wilms' tumor. Pelviureteric junction tissues were confirmed to be free of tumor invasion by histology. Immunohistochemistry with c-kit antibody was performed to detect the expression of ICC in specimens of the 2 groups. Quantitative analysis was made using image analysis technique and statistical analysis was carried out.

Results

Immunoreactivity to ICC was predominantly detected in the muscle layers of PUJ. The mean area of ICC expression in the PUJO group was 14.86 ± 1.37 × 10⁴μm² , which was lower than that in the control group (16.80 ± 1.68) × 10⁴μm² (P < .01). The mean density of ICC expression in the PUJO was 0.207 ± 0.020, which was also lower than that in the control group (0.262 ± 0.026) (P < .05).

Conclusions

A reduction of the number of ICC may play an important role in the etiology and pathogenesis of PUJO.     

肝外胆管梗阻性疾病的MRCP诊断价值

目的探讨磁共振胆胰管成像技术对肝外胆管梗阻性疾病的诊断价值。方法对52例经病理或随访证实的肝外胆管梗阻性疾病患者的MRCP资料进行回顾性分析,总结良恶性肝外胆管梗阻的不同MRCP表现。结果52例中,MRCP均能够准确测定胆管扩张程度和梗阻的水平,其中良性梗阻27例,MRCP主要表现为胆管均匀扩张及逐渐狭窄,肝外胆管较肝内胆管扩张明显,肝内胆管呈“枯树枝状”;恶性梗阻25例,MRCP主要表现为胆管截断,胆胰管扩张,出现“双管征”,肝内外胆管扩张一致。恶性梗阻患者的肝内外胆管扩张程度明显大于良性梗阻者。结论MRCP作为一种无创的影像检查方法,对肝外胆管梗阻性疾病的诊断具有较高准确性,     

放射性镍钛合金内支架治疗晚期恶性胆管梗阻

目的 探讨放射性镍钛记忆合金胆道内支撑架对晚期恶性胆管梗阻的治疗效果。方法 通过对比放射性镍钛合金胆道内支架，Wallstent金属内支架，胆肠吻合术在减黄效果，术后并发症，生存时间等方面 疗效，研究放射性镍钛合金胆道内支架在治疗晚期恶性胆管梗阻方面的优越性。结果 三组在减黄效果上无明显差别，但放射性镍钛合金内支架组在无症状生存时间及总的生存时间方向较另两组长，其术后并发症较另两组明显少（P＜0．01），复发率低，而且病人痛苦小，生存质量明显得到提高。结论 放射性镍钛合金胆道支架既能解决晚期恶性胆管梗阻病人因胆道梗阻所致黄疸带来的痛苦，又能有效抑制肿瘤细胞的生长，对于延长病人生存时间，提高生存质量具有切实疗效。     

Protective effect of polyunsaturated phosphatidylcholine on liver damage induced by biliary obstruction in rats

Background/purpose: Persistent inflammatory response secondary to congenital or acquired biliary choleastasis plays an important role in the pathophysiology of hepatic tissue damage. The polyunsaturated fatty acids (PUFA) have been shown to suppress the inflammatory reactions in vivo and in vitro. PUFA has been shown also to protect againts various types of experimental liver damage in animal models and isolated hepatocytes. Therefore, the aim of this study was to investigate the protective effect of PUFA administration on liver damage using the rat chronic biliary obstruction model.Methods: Swiss albino rats of either sex were divided into 4 groups as follows: control group (group 1, 10 rats); rats with sham operation and treated with saline group 2, 10 rats); rats with biliary obstruction (group 3, 15 rats); and polyunsaturated phophatidylcholine (PPC)-treated rats with biliary obstruction (Group 4, 15 rats). Biliary obstruction was induced by double ligation and division of the common bile duct. PUFA treatment was started 2 weeks later from biliary obstruction in doses of 50 mg/d per rat and continued for 2 weeks. All animals were killed after 4 weeks of common bile duct ligation or sham operation. Liver damage and cholestasis were determined by biochemical and histologic examinations.Results: The data showed a decrease in plasma bilirubin level (both conjugated and unconjugated) and liver enzyme levels (AST, ALT, AP, GGT, 5′-NT) in group 4, when compared with group 3 (P < .05). Tissue levels of malondialdehyde (MDA) in group 4 was 20.00 ± 2.93 compared with that in group 3, 27.12 ± 2.96 (P < .05). Administration of PUFA to the biliary obstructed rats resulted in inhibition of collagen accumulation (P < .05) and ductal proliferation (P < .05).Conclusions: PUFA reduced liver damage, ductular proliferation, and fibrosis in biliary obstructed rats. These effects suggest that it might be a useful agent to preserve liver function in patients with biliary obstruction such as biliary atresia.     

干细胞因子促进Cajal间质细胞恢复的研究

目的 观察阻断c-Kit信号通路致Caja1间质细胞(ICC)缺失,而c-Kit的配体干细胞因子(SCF)能否促进其恢复.方法 将新出生的小鼠于腹腔内隔天注射c -Kit抗体(ACK2) 100μg,共5次,建立ICC缺失模型,再于第10天腹腔内注射SCF,连续注射5d后观察空肠电节律并通过免疫组织化学法观察Cajal间质细胞的恢复；采用逆转录-聚合酶链反应(RT-PCR)、Western blot法检测c-Kit基因转录和表达.结果 当c-Kit受体被阻断后,空肠ICC几乎缺失且慢波消失；SCF腹腔注射给药可使ICC缺失小鼠空肠慢波频率明显增快(7.83±1.34)次/min(P ＜0.01),振幅也有加大(0.044±0.009) mV(P ＜0.05)；而且肌间神经丛区ICC增多,c-Kit mRNA及c-Kit蛋白表达也上调.结论 c-Kit与其配体SCF结合所启动的信号途径对ICC的生存及表型维持至关重要.阻断c-Kit的作用后,ICC会出现缺失.通过给予外源性SCF刺激后,缺失的ICC部分恢复,且这种恢复与c-Kit表达水平的上调有关.     

球囊扩张法治疗损伤性胆管狭窄的前瞻性研究

目的 探讨球囊扩张法治疗损伤性胆管狭窄(TBS)修复术后再狭窄的效果.方法 将1997年2月至2007年2月成都军区总医院收治的81例TBS患者按随机数字表法分成2组:对照组40例,采用传统胆肠吻合术,术后T管支撑6个月;球囊组41例,方法同对照组,同时在吻合口置人球囊,并逐渐扩张肝门部胆管空肠吻合口及上方胆管.两组疗效比较采用x2检验.结果 对照组7例失败(18%),其中6例改用球囊扩张治疗后5例获得成功.球囊组2个月内吻合口径达到12 mill,再维持1个月后将T管和球囊拔出,仅1例失败(2%).两组比较差异有统计学意义(x2=5.10,P＜0.05).结论 球囊扩张法能大大提高TBS修复术的疗效.