幽门螺杆菌感染患儿胃黏膜紧密连接蛋白表达的变化及其意义

发布时间：2018-06-23 03:38

  本文选题：幽门螺杆菌 + 紧密连接蛋白　； 参考：《浙江大学》2013年硕士论文

【摘要】：研究背景： 胃黏膜具有阻止各种物理和化学物质损伤的防御机制,包括黏液-碳酸氢盐屏障、黏膜屏障、黏膜血流和上皮细胞更新。胃黏膜屏障主要由上皮细胞及细胞间连接组成,可有效防止H+的反向弥散,阻止大分子入侵,是防止有害微生物经胃侵入机体的第一道防线。细胞间连接是由紧密连接、黏附连接、缝隙连接和桥粒等组成。一旦紧密连接受损,上皮细胞间的通透性增加,细菌、内毒素等就可通过紧密连接进入机体,这与多种疾病的发生发展有关。幽门螺杆菌(helicobacter pylori, Hp)感染与慢性胃炎、消化性溃疡、胃腺癌和黏膜相关性淋巴组织淋巴瘤密切相关,但其发病机制尚不清楚。研究表明,Hp产生的毒素和有毒素作用的酶能破坏胃黏膜屏障,还能使机体产生炎症和免疫反应,导致疾病的发生。流行病学资料显示,50%以上Hp相关疾病患者是在儿童期受到感染,Hp与儿童慢性胃炎、十二指肠炎、消化性溃疡的发生发展密切相关。目前认为,Hp感染后胃上皮屏障功能的受损在Hp相关性胃病的发病中起着重要的作用,有关儿童Hp感染对胃上皮屏障功能的影响尚不明了。因此,进一步研究儿童Hp感染后胃黏膜紧密连接蛋白表达及其变化,对于了解Hp相关性胃病的发病机制有着重要的临床意义。 目的： 探讨Hp感染患儿胃黏膜紧密连接蛋白如咬合蛋白(occludin)、闭合蛋白(claudin)、闭合小环蛋白(ZO-1)、上皮型钙黏蛋白(E-cadherin)和μ连环蛋白(β-catenin)表达的变化及其临床意义。 方法： 收集2011年5月-11月因“恶心、呕吐、腹痛、腹胀或反酸、黑便”等消化道症状在浙江大学医学院附属儿童医院胃肠镜室接受胃镜检查,所有研究均与家长签署知情同意书,并符合医学伦理原则。胃镜检查时在患儿胃窦部取3块活检组织,一块用于快速尿素酶检测,一块固定于10%福尔马林送检病理,一块液氮冻存后.80℃低温冰箱中保存。胃黏膜紧密连接蛋白mRNA水平的测定采用RT-PCR法,其蛋白质水平的测定采用Western blot法；E-cadherin和β-catenin在胃黏膜上皮的定位和半定量测定采用免疫组织化学法。 结果： 临床病理资料完整的患儿共41例,其中男28例,女13例,年龄3.5-16岁。幽门螺杆菌尿素酶及病理检测Hp阳性27例,Hp阴性14例。Hp阳性组中消化性溃疡者13例,非溃疡者14例。在Hp感染患儿的胃黏膜组织中无论有溃疡组及无溃疡组,ZO-1、E-cadherin、β-catenin mRNA表达水平低于Hp阴性组,差异有统计学意义(P均0.01)；claudin-4在Hp阳性溃疡组表达明显升高,差异有统计学意义(P0.01)；而occludin在三组间差异没有统计学意义。免疫组织化学结果显示,Hp阳性组患儿的E-cadherin、β-catenin蛋白表达水平低于阴性对照组,差异有统计学意义(P分别0.01和0.05)。 结论： Hp感染患儿胃黏膜紧密连接蛋白ZO-1、E-cadherin、β-catenin表达降低,消化性溃疡患儿claudin-4表达增加,提示儿童Hp感染后胃上皮屏障功能受损可能是Hp相关性胃病的主要致病机制。
[Abstract]:Research background:
The gastric mucosa has a defense mechanism that prevents damage to various physical and chemical substances, including the mucous - bicarbonate barrier, the mucosal barrier, the mucosal blood flow and the epithelial cell renewal. The gastric mucosal barrier is composed mainly of epithelial cells and intercellular connections, which can effectively prevent the reverse dispersion of H+, prevent the invasion of large molecules, and prevent the harmful microorganism from invading through the stomach. The first line of defense in the body. Intercellular connections are composed of close connections, adhesion connections, gap junctions and bridging particles. Once the close connections are damaged, the permeability between the epithelial cells increases, bacteria, endotoxin and so on can enter the body through close connection, which is related to the development of a variety of diseases. Helicobacter pylori, H P) infection is closely related to chronic gastritis, peptic ulcers, gastric adenocarcinoma and mucosa associated lymphoid tissue lymphoma, but its pathogenesis is still unclear. Studies have shown that the toxins produced by Hp and enzymes that have toxic effects can destroy the gastric mucosal barrier, and can also cause inflammation and immune response to the disease. Epidemiological data have been shown to show. More than 50% of the patients with Hp related diseases are infected in childhood, and Hp is closely related to the development of chronic gastritis, duodenitis and peptic ulcers in children. It is now believed that the damage of the gastric epithelial barrier function after Hp infection plays an important role in the pathogenesis of Hp related gastropathy, and related to the function of Hp infection to the gastric epithelial barrier in children. Therefore, further study of the expression and changes of the close connexin in the gastric mucosa after Hp infection in children has important clinical significance for understanding the pathogenesis of Hp related gastropathy.
Objective:
The changes and clinical significance of the expression of close connexin (occludin), closed protein (claudin), closed small cyclic protein (ZO-1), epithelial Calc mucin (E-cadherin) and muon catenin (beta -catenin) in the gastric mucosa of children with Hp infection were investigated.
Method:
The digestive tract symptoms, such as "nausea, vomiting, abdominal pain, abdominal distention, or anti acid, and stool", were collected in the gastroenteroscopy room of the children's Hospital Affiliated to Zhejiang University Medical College, Zhejiang University, in May 2011. All the studies signed the informed consent with the parents and were in accordance with the principles of medical ethics. In the gastroscopy, 3 biopsy tissues were taken in the antrum of the children's stomach. Used for rapid urease detection, one was fixed in 10% formalin to be examined by 10% formalin, and one piece of liquid nitrogen was stored in cryogenic refrigerator after cryopreservation. RT-PCR method was used to determine the level of close connexin mRNA in gastric mucosa. The protein level was determined by Western blot; E-cadherin and beta -catenin were located and semi quantified in the gastric mucosa epithelium. The immuno histochemical method was adopted.
Result锛，

本文编号：2055588