Vorobjova Tamara, Raikkerus Helerin, Kadaja Lumme, Talja Ija, Uibo Oivi, Heilman Kaire, Uibo Raivo
Department of Immunology, Institute of Biomedicine and Translational Medicine, University of Tartu, Ravila 19, 51014, Tartu, Estonia.
Department of Pathophysiology, Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia.
Dig Dis Sci. 2017 Feb;62(2):358-371. doi: 10.1007/s10620-016-4403-z. Epub 2016 Dec 19.
Impaired intestinal integrity, including increased permeability of the small bowel mucosa, has been shown in patients with celiac disease (CD) as well as with type 1 diabetes (T1D). Zonulin (ZO, pre-haptoglobin), a tight junction regulator, plays a particular role in the regulation of intestinal barrier function and in the pathogenesis of the above-mentioned diseases.
To investigate whether enteroviruses (EVs) and immunoregulatory cells are associated with intestinal permeability in patients with CD alone and with coexistent T1D.
Altogether 80 patients (mean age 10.68 ± 6.69 years) who had undergone small bowel biopsy were studied. Forty patients with functional dyspepsia and normal small bowel mucosa formed the control group. The circulating ZO level in sera was evaluated using ELISA. The densities of EV, FOXP3+ regulatory T cells (Tregs), indoleamine 2,3-dioxygenase (IDO+) dendritic cells (DCs) and glutamic acid dexarboxylase (GAD)65+ cells in small bowel mucosa were investigated by immunohistochemistry. The expression analysis of FOXP3, tight junction protein 1 (TJP1), gap junction (GJA1), IDO and CD103 genes was evaluated by real-time PCR.
The ZO level was higher in CD patients compared to subjects with a normal small bowel mucosa, particularly in those with Marsh IIIc atrophy (p = 0.01), and correlated with the density of EV (r = 0.63; p = 0.0003) and IDO+ DCs (r = 0.58; p = 0.01) in the small bowel mucosa. The density of GAD65+ epithelial cells was correlated with the density of EV (r = 0.59; p = 0.03) and IDO+ DCs (r = 0.78; p = 0.004) in CD patients. The relative expression of FOXP3 mRNA in the small bowel mucosa tissue was significantly higher in patients with CD, compared to subjects with a normal mucosa, and correlated with the density of EV (r = 0.62; p = 0.017) as well as with the relative expression of IDO mRNA (r = 0.54; p = 0.019).
The CD is associated with elevation of the circulating ZO level, the value of which correlates with the density of EV in CD patients with severe atrophic changes in the small bowel mucosa, particularly in cases of concomitant T1D. The CD is also characterized by the close relationship of the density of GAD65+ epithelial cells with the EV, ZO level and IDO+ DCs.
乳糜泻（CD）患者以及1型糖尿病（T1D）患者均出现肠完整性受损，包括小肠黏膜通透性增加。闭合蛋白（ZO，前触珠蛋白）作为一种紧密连接调节因子，在肠道屏障功能调节及上述疾病的发病机制中发挥着特殊作用。
研究肠道病毒（EVs）和免疫调节细胞是否与单纯CD患者以及合并T1D患者的肠道通透性相关。
共研究了80例接受小肠活检的患者（平均年龄10.68±6.69岁）。40例功能性消化不良且小肠黏膜正常的患者作为对照组。采用酶联免疫吸附测定法（ELISA）评估血清中循环闭合蛋白水平。通过免疫组织化学研究小肠黏膜中EV、叉头框P3+调节性T细胞（Tregs）、吲哚胺2,3-双加氧酶（IDO+）树突状细胞（DCs）和谷氨酸脱羧酶（GAD）65+细胞的密度。通过实时聚合酶链反应（PCR）评估叉头框P3、紧密连接蛋白1（TJP1）、缝隙连接蛋白（GJA1）、IDO和CD103基因的表达分析。
与小肠黏膜正常的受试者相比，CD患者的闭合蛋白水平更高，尤其是马什IIIc级萎缩患者（p = 0.01），且与小肠黏膜中EV的密度（r = 0.63；p = 0.0003）和IDO+ DCs的密度（r = 0.58；p = 0.01）相关。CD患者中GAD65+上皮细胞的密度与EV的密度（r = 0.59；p = 0.03）和IDO+ DCs的密度（r = 0.78；p = 0.004）相关。与黏膜正常的受试者相比，CD患者小肠黏膜组织中叉头框P3信使核糖核酸（mRNA）的相对表达显著更高，且与EV的密度（r = 0.62；p = 0.017）以及IDO mRNA的相对表达（r = 0.54；p = 0.019）相关。
CD与循环闭合蛋白水平升高相关，其水平与小肠黏膜有严重萎缩性改变的CD患者（尤其是合并T1D的病例）中EV的密度相关。CD的特征还在于GAD65+上皮细胞的密度与EV、闭合蛋白水平和IDO+ DCs密切相关。
