酸敏感离子通道影响食管酸敏感性的机制研究

发布时间：2018-06-09 20:20

本文选题：胃食管反流病 + 内脏痛觉高敏感　；参考：《第二军医大学》2014年硕士论文

【摘要】：研究背景：胃食管反流病（gastro esophageal reflux disease，GERD)发病机制错综复杂，，研究显示部分GERD患者食管的酸暴露正常，有些有过长时间的酸暴露时间，随着进一步的研究，逐渐认识到不能简单地将GERD归结为反流物对食管粘膜的化学性刺激直接造成诱发组织损伤，使神经末梢暴露于酸环境。目前炎症和内脏高敏感（visceralhypersensitivity,VH）在GERD的发病过程中起到关键作用。GERD患者食管粘膜中的炎症介质可降低酸敏感离子通道(acid-sensing ion channels,ASICs)受体的信号转导阈值，从而介导外周致敏，引起酸痛觉高敏现象。ASICs各亚基在哺乳动物各器官组织中的分布具有特异性，对细胞外pH的敏感性不同。目前明确ASIC1、ASIC2和ASIC3均表达于食管及其支配神经，ASICs的不同亚型可能在GERD的发病过程中起截然不同的作用。其中ASIC1主要介导对酸和机械性刺激的抑制性反应，ASIC3主要是介导对酸刺激和机械扩张刺激的敏感性反应，同时ASIC3还参与酸刺激后的炎症反应过程。但酸敏感离子通道亚基在食管痛觉高敏感形成和维持中机制未明。研究目的：本研究旨在观察不同ASICs受体亚基在GERD患者食管粘膜的表达变化并评估不同ASICs亚基表达变化与患者食管痛觉阈值改变的相关性。研究方法招募明显反流、烧心等症状的患者填写GERD调查问卷。GERD-Q评分大于6分的患者进行电子胃镜检查，根据患者临床表现(包括临床症状、质子泵抑制剂治疗反应等)、电子内镜和病理检查结果，将其分实验组：反流性食管炎（RE）患者胃镜下Los-Angeles分级为B级及以上的在患处取材（A组）并在患处旁取材（病例自身对照组-D组）；Los-Angeles分级为A级的在患处取材（B组）；对照组以齿状线上3cm处为标准，留取受试者食道粘膜活检标本。16位正常志愿者组成无症状对照组（C组）。使用免疫荧光及实时定量聚合酶链反应（Real-time PCR）的方法检测GERD患者食管粘膜ASIC1和ASIC3受体亚基的表达改变。评价GERD患者ASIC1＆3与典型症状和炎症程度的相关性。研究结果胃镜下轻度食管炎可见食管粘膜稍红，严重的可有糜烂和溃疡。组织学的改变主要以慢性炎症为表现，鳞状上皮增生，粘膜固有层乳头向上皮腔面延伸，浅层毛细血管扩张。固有层内以淋巴细胞浸润为主，血管化的上皮及其出血的小灶，粘膜下层小静脉扩张，毛细血管增多。对照组食管粘膜组织未见明确病理学改变。 Real Time-PCR检测结果：ASIC1mRNA相对表达量为A组36.527±34.912，B组21.242±14.325，C组4.404±2.938，D组5.413±4.215。A组及B组均较C组及D组明显升高，P＜0.05，A组及B组间与C组及D组间不存在统计学差异； ASIC3mRNA相对表达量为A组5.200±4.343， B组1.137±0.660，C组0.352±0.164，D组0.419±0.349。A组较B组、C组及D组升高，B组较C组及D组升高，P＜0.05，C组及D组间不存在统计学差异。免疫荧光检测结果：A组ASIC1积分光密度值为0.107±0.012，B组ASIC1积分光密度值为0.100±0.018，C组ASIC1积分光密度值为0.048±0.006，D组ASIC1积分光密度值为0.053±0.007。A组及B组均较C组及D组明显升高，P＜0.01，A组及B组间与C组及D组间不存在统计学差异；A组ASIC3积分光密度值为0.075±0.006， B组ASIC3积分光密度值为0.075±0.004，C组ASIC3积分光密度值为0.055±0.007，D组ASIC3积分光密度值为0.055±0.007。A组及B组均较C组及D组明显升高，P＜0.01，A组及B组间与C组及D组间不存在统计学差异。ASIC1＆3的表达与RE的炎症程度无明显相关性，ASIC1＆ASIC3与典型症状烧心呈正相关，Pearson相关系数分别为0.675、0.571，ASIC1＆ASIC3与典型症状胸痛呈正相关，Pearson相关系数分别为0.636、0.611，与反流无明显相关性。结论 GERD患者食管粘膜ASIC1和ASIC3的表达发生了改变，这表明ASIC1和ASIC3在食管痛觉高敏感形成和维持中发挥重要作用。
[Abstract]:Research background:
The pathogenesis of gastro esophageal reflux disease (GERD) is complicated. The study shows that the acid exposure of the esophagus is normal in some GERD patients and the time of acid exposure for long time. With further study, it is gradually realized that the chemical stimulation of the esophagus mucous membrane can not be simply combined with GERD as a reflux. At present, inflammation and visceral Gao Mingan (visceralhypersensitivity, VH) play a key role in the pathogenesis of GERD, and the inflammatory mediators in the esophageal mucosa of.GERD patients can reduce the signal transduction threshold of the acid sensitive ion channel (acid-sensing ion channels, ASICs) receptor, and thus mediate the signal transduction threshold of the acid sensitive ion channel (acid-sensing ion channels, ASICs). The sensitivity of the.ASICs subunits in the organs and tissues of the mammalian organs is specific, and the sensitivity to the extracellular pH is different. At present, it is clear that ASIC1, ASIC2 and ASIC3 are all expressed in the esophagus and its dominant nerve, and the different subtypes of ASICs may play a very different role in the pathogenesis of GERD. ASIC1 mainly mediates inhibitory reactions to acid and mechanical stimuli, and ASIC3 is mainly mediated by sensitive reactions to acid stimulation and mechanical dilatation, and ASIC3 is also involved in the process of inflammatory response after acid stimulation.
The purpose of the study is:
The purpose of this study was to observe the changes in the expression of different ASICs receptor subunits in the esophageal mucosa of patients with GERD and to evaluate the correlation between the changes in the expression of different ASICs subunits and the changes of the esophageal pain threshold.
research method
Patients who recruited obvious reflux, heart burning and other symptoms filled in the GERD questionnaire.GERD-Q score greater than 6 for electronic gastroscopy, according to the clinical manifestations of the patients (including clinical symptoms, proton pump inhibitors treatment, etc.), electronic endoscopy and pathological results, and the experimental group: reflux esophagitis (RE) patients under the gastroscope Los-An Geles was classified as grade B and above in the affected area (group A) and taken from the affected part (case control group -D); Los-Angeles was classified as class A in the affected area (group B); the control group took 3cm at the dentate line as the standard, and the normal volunteers of.16 position of the esophagus mucosa biopsy specimen of the subjects were composed of the asymptomatic control group (C group).
The expression of ASIC1 and ASIC3 receptor subunits in the esophageal mucosa of GERD patients was detected by immunofluorescence and real-time quantitative polymerase chain reaction (Real-time PCR). The correlation between the atic 1 & 3 and the typical symptoms and the degree of inflammation in the GERD patients was evaluated.
Research results
Mild esophagitis under gastroscope shows a slight reddish mucous membrane of the esophagus, severe erosion and ulcers. The histological changes are mainly characterized by chronic inflammation, squamous epithelium hyperplasia, the extension of the lamina propria papillae to the epithelial cavity, and the dilatation of the superficial capillaries. There was no obvious pathological changes in the esophageal mucosa of the control group.
The results of Real Time-PCR detection: the relative expression of ASIC1mRNA was 36.527 + 34.912 in group A, 21.242 in group B and 4.404 in C in group C, and in group 5.413 + 4.215.A and B group of D group was significantly higher than that in C group and D group, and P < 0.05. Group 0.352 + 0.164, group D 0.419 + 0.349.A group was higher than group B, group C and D group, B group was higher than C group and D group, P < 0.05, C group and D group had no statistical difference.
The results of immunofluorescence detection: A group ASIC1 integral light density value is 0.107 + 0.012, B group ASIC1 integral light density value is 0.100 + 0.018, C group ASIC1 integral light density value is 0.048 + 0.006, D group ASIC1 integral light density value is 0.053 + 0.007.A group and B group is higher than C group and D group, < 0.01. The optical density of ASIC3 integral in A group was 0.075 + 0.006, B group ASIC3 integral light density value was 0.075 + 0.004, C group ASIC3 integral light density value was 0.055 + 0.007, ASIC3 integral light density value of D group was 0.055 + 0.007.A group and B group was significantly higher than C group and D group. There was no significant correlation between the expression of RE and the degree of inflammation of the typical symptoms. The correlation coefficient of Pearson was 0.675,0.571, and the atic 1 & AC 3 was positively correlated with the typical symptoms of chest pain, and the correlation coefficient of Pearson was 0.636,0.611, and there was no obvious correlation with reflux.
conclusion
The expression of ASIC1 and ASIC3 in esophageal mucosa of GERD patients has changed. This indicates that ASIC1 and ASIC3 play an important role in the formation and maintenance of esophageal hyperalgesia.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R571

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