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成年豚鼠小肠壁内Cajal细胞再生及细胞网络再形成实验研究

发布时间:2018-02-24 16:08

  本文关键词: Cajal 细胞 KIT 受体 BrdU 再生 干细胞因子 神经 平滑肌细胞 出处:《第三军医大学》2005年硕士论文 论文类型:学位论文


【摘要】:Cajal 细胞(Interstitial cells of Cajal, ICCs) 作为一种特殊的间质细胞,以细胞网络的形式分布于整个胃肠道。近二十余年,国内外学者通过对胃肠壁内ICCs 的形态特征、超微结构、起源及生理功能等的一系列研究表明:ICCs 具有产生自发性电慢波、参与神经信息传递等功能,在调节胃肠道平滑肌运动中发挥重要作用。 动物实验发现:胃肠壁内ICCs 减少、细胞网络缺如的大鼠和小鼠,胃肠运动自主节律性消失,呈扩张性肠麻痹状态,胃和肠的内容物反流,提示ICCs 减少及其细胞网络完整性的破坏可能与上述现象的发生有关。临床可见胃肠道手术后患者常常出现一过性肠麻痹和胃肠蠕动运动功能消失等假性肠梗阻症状,推测可能与ICCs 细胞网络断裂和手术部位ICCs 的表型(phenotype)发生急性变化(不能被其特异性标志物所染色),丧失正常功能有关。近年临床研究亦发现ICCs 与一些胃肠运动功能障碍性疾病:例如贲门失弛缓症、慢传输型便秘、假性肠梗阻、先天性巨结肠等密切相关。尽管上述疾病的发生机制尚不清楚,但均有一个共同的特点:病变部位胃肠壁内ICCs数量呈不同程度减少,甚至缺如,ICCs 彼此间及其与平滑肌之间不能形成完整的细胞网络。但因对上述疾病的发病机理及ICCs 减少、细胞网络完整性被破坏的原因等尚不清楚,一直是临床治疗的难题。 设想如果能够阐明成年动物消化管壁内ICCs 损伤丢失后,是否通过再生、细胞突起生长延长、重新形成细胞网络的调控机制,势必可为临床相关疾病的治疗及腹部外科术后胃肠运动功能的恢复提供有益的思路。因此,本实验利用外科手术的方法,模拟术后胃肠运动功能障碍,制作小肠壁内ICCs 受损及细胞网络完整性被破坏的动物模型,结合免疫组化染色、双重标记技术、透射电镜观察等方法,动态分析小肠半横断吻合术后ICCs 的变化。该模型不仅能够使我们了解小肠术后ICCs 的变化,同时也有助于阐明成年动物肠壁内ICCs 是否具有分裂增殖能力、随着时间的经过ICCs 的突起能否通过生长延长重新形成细胞网络,及其平滑肌细胞和神经对此过程的影响等问题,为治疗胃肠运动功能障碍性疾病开辟新的思路和方法。 主要结果如下: 根据吻合断端的特点,将其分为3 区域:Ⅰ区位于肠壁横断末端,平滑肌层因回缩而稍增厚,该区突出于肠腔;Ⅱ区是紧靠Ⅰ区正常侧的平滑肌层,是吻合部位,由于缝线张力的牵拉作用使该区肠壁肌层变薄;Ⅲ区是Ⅰ区和Ⅱ区浆膜面与吻合中线间的区域。
[Abstract]:As a special mesenchymal cell, Cajal cells are distributed in the whole gastrointestinal tract in the form of cellular network. In the past 20 years, the morphological features and ultrastructure of ICCs in the wall of stomach and intestine were analyzed by domestic and foreign scholars. A series of studies on the origin and physiological function of ICCs have shown that ICCs have the functions of producing spontaneous slow waves and participating in the transmission of neural information. They play an important role in regulating the movement of gastrointestinal smooth muscle. Animal experiments showed that in rats and mice with decreased ICCs in gastrointestinal wall and absent cellular network, the autonomic rhythm of gastrointestinal motility disappeared, showing dilated intestinal paralysis, and reflux of the contents of stomach and intestine. The results suggest that the decrease of ICCs and the destruction of cellular network integrity may be related to the occurrence of these phenomena. The clinical symptoms of pseudo-intestinal obstruction such as transient intestinal paralysis and disappearance of gastrointestinal peristaltic motor function are often found in patients with gastrointestinal tract operation. It is speculated that the acute changes of ICCs cell network breakage and phenotype of ICCs at the site of operation (which can not be stained by its specific markers) may be related to the loss of normal function. In recent years, clinical studies have also found that ICCs and some gastrointestinal motility have been involved. Disorders such as achalasia, Slow transit constipation, pseudo-intestinal obstruction and Hirschsprung's disease are closely related. Although the pathogenesis of these diseases is not clear, they all have a common characteristic: the number of ICCs in gastrointestinal wall decreases in varying degrees. Even without ICCs and smooth muscle cells can not form a complete network, but the pathogenesis of the disease and the reduction of ICCs, the reasons for the destruction of the integrity of the cell network are still unclear, which has been a difficult problem in clinical treatment. Imagine that if we can clarify the regulatory mechanisms of ICCs damage loss in the digestive walls of adult animals, whether it is possible to regenerate, prolong the growth of the cell processes and reform the cellular network. It will provide useful ideas for the treatment of clinical related diseases and the recovery of gastrointestinal motility after abdominal surgery. The animal model of damaged ICCs and damaged cellular network integrity in the small intestine wall was made by immunohistochemical staining, double labeling technique, transmission electron microscopy and other methods. Dynamic analysis of the changes of ICCs after semi-transverse anastomosis of small intestine. This model can not only help us understand the changes of ICCs after intestinal surgery, but also help us to clarify whether the ICCs in the intestinal wall of adult animals has the ability to divide and proliferate. Over time, whether the processes of ICCs can reform the cellular network through growth prolongation, and the effects of smooth muscle cells and nerves on this process, will open up new ideas and methods for the treatment of gastrointestinal motility disorders. The main results are as follows:. According to the characteristics of anastomotic end, it is divided into three regions: zone I is located at the end of the transverse section of the intestinal wall, the smooth muscle layer is slightly thickened by the retraction of the smooth muscle layer, and the region protrudes out in the intestinal cavity, and area 鈪,

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