Proteinase-Activated Receptor-2 in the Pathogenesis of Gastroesophageal Reflux Disease
Proteinase-Activated Receptor-2 in the Pathogenesis of Gastroesophageal Reflux Disease
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DOI：
10.1038/ajg.2010.265
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发表时间：
2010-09-01
影响因子：
9.8
通讯作者：
Malfertheiner, Peter
中科院分区：
文献类型：
--
作者：
Kandulski, Arne;Wex, Thomas;Malfertheiner, Peter
目的：蛋白水解酶激活受体-2(PAR-2)由丝氨酸蛋白酶激活，具有促炎和神经炎性作用。它被认为改变了跨上皮阻力，并介导了内脏高敏感性。方法：将123例胃食道反流病患者分为糜烂性反流病(n=50)、非糜烂性反流病(n=46)和反流阴性患者(n=27)。分别根据洛杉矶分类和改良的Ismail-Beigi标准进行内窥镜检查和组织病理学检查。用定量逆转录聚合酶链式反应和免疫组织化学方法检测PAR-2的表达。采用定量RT-PCR方法检测IL-8基因表达水平，并与PAR-2表达水平进行相关性分析。体外培养食管鳞状细胞系(KYSE150、KYSE450)，调节不同的pH值(7.0、6.0、5.0)，并加入胆汁酸和PAR-2活化肽(SLIGKV-NH2)。结果：PAR-2基因表达上调7~10倍(P<；0.0001)，并与IL-8表达及组织形态改变(细胞间隙扩大、乳头延长、基底细胞增生)呈正相关。免疫组织化学结果显示，与对照组相比，胃肠道反流性疾病组各上皮层均有PAR-2的强阳性表达(P=0.0005)。体外研究表明，酸化介质(P<；0.01)可诱导食道鳞状细胞PAR-2基因表达1.5-20倍，但不能被额外的胆汁酸诱导。PAR-2的激活导致IL-8的表达和分泌。结论：本研究为PAR-2介导的途径在GERD及GERD相关的粘膜改变和炎症改变的发病机制中的功能重要性提供了证据。
OBJECTIVES: The proteinase-activated receptor-2 (PAR-2) is activated by serine proteases and has been demonstrated to induce proinflammatory and neuroinflammatory effects. It is considered to alter transepithelial resistance and mediates visceral hypersensitivity. This study aimed to evaluate the expression of PAR-2 in human esophageal mucosa of patients with gastroesophageal reflux disease (GERD) in relation to mucosal alterations.METHODS: The study included 123 patients with GERD stratified to erosive reflux disease (n = 50), non-erosive reflux disease (n = 46), and reflux-negative patients as controls (n = 27). Endoscopic and histopathological characterization was performed according to the Los Angeles classification and modified Ismail-Beigi criteria, respectively. PAR-2 expression was analyzed by quantitative reverse transcription (RT)-PCR and immunohistochemistry. The gene expression levels of interleukin (IL)-8 were determined by quantitative RT-PCR and correlated to PAR-2 expression in each patient. Performing in vitro studies, esophageal squamous cell lines (KYSE 150, KYSE 450) were incubated, adjusted to different pH (7.0, 6.0, and 5.0), and exposed to bile acids and PAR-2-activation peptide (SLIGKV-NH2).RESULTS: PAR-2 gene expression was 7- to 10-fold upregulated (P < 0.0001) in the mucosa of patients with GERD and correlated positively with IL-8 expression and with histomorphological alterations (dilated intercellular spaces, papillary elongation, basal cell hyperplasia (BCH); P < 0.01). Immunohistochemistry showed an intense staining of PAR-2 throughout all epithelial layers in patients with GERD compared with controls (P = 0.0005). In vitro studies revealed a 1.5- to 20-fold induction of PAR-2 gene expression in esophageal squamous cells by acidified medium (P < 0.01), but not by additional bile acids. The activation of PAR-2 leads to expression and secretion of IL-8.CONCLUSIONS: This study provides evidence of the functional importance of PAR-2-mediated pathways in the pathogenesis of GERD and GERD-associated mucosal alterations and inflammatory changes.