Evaluation of Porcine Intestinal Epitheliocytes as an In vitro Immunoassay System for the Selection of Probiotic Bifidobacteria to Alleviate Inflammatory Bowel Disease
Evaluation of Porcine Intestinal Epitheliocytes as an In vitro Immunoassay System for the Selection of Probiotic Bifidobacteria to Alleviate Inflammatory Bowel Disease
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DOI：
10.1007/s12602-020-09694-z
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发表时间：
2020-08
影响因子：
4.9
通讯作者：
Nana Sato;M. Yuzawa;Md. Aminul;M. Tomokiyo;L. Albarracín;V. García-Castillo;Wakako Ideka-Ohtsubo;N. Iwabuchi;J. Xiao;A. García-Cancino;J. Villena;H. Kitazawa
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文献类型：
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作者：
Nana Sato;M. Yuzawa;Md. Aminul;M. Tomokiyo;L. Albarracín;V. García-Castillo;Wakako Ideka-Ohtsubo;N. Iwabuchi;J. Xiao;A. García-Cancino;J. Villena;H. Kitazawa
使用允许有效选择具有免疫调节特性的益生菌候选物的体外系统可以显着减少实验动物的使用。在这项工作中，我们产生了一种基于猪肠上皮细胞（PIE）和葡聚糖硫酸钠（DSS）管理的体外免疫测定系统，这可能有助于选择和表征用于炎症性肠病（IBD）患者的潜在益生菌菌株。我们的策略基于两个基本支柱：一方面，PIE细胞通过附着到相邻细胞并有效地产生炎症反应而形成单层的能力，另一方面，使用两种益生菌双歧杆菌菌株，其特征在于其免疫调节能力，特别是在小鼠IBD模型和患者中。我们的研究结果表明，DSS给药可改变PIE细胞在体外产生的上皮屏障，并诱导强效炎症反应，其特征在于几种炎症因子的表达水平增加，包括TNF-α、IL-1α、CCL 4、CCL 8、CCL 11、CXCL 5、CXCL 9、CXCL 10、SELL、SELE、EPCAM、VCAM、NCF 2和SAA 2。此外，我们证明了短双歧杆菌M-16 V和长双歧杆菌BB 536能够调节C-jun N-末端激酶（JNK）细胞内信号通路，减少DSS诱导的体外上皮屏障改变，并以应变依赖性方式差异调节炎症反应。我们在PIE细胞中的体外研究结果与先前在动物模型和IBD患者中的研究之间的良好相关性显示了我们的系统以有效方式选择新益生菌候选物的潜在价值。
The use of in vitro systems that allow efficient selection of probiotic candidates with immunomodulatory properties could significantly minimize the use of experimental animals. In this work, we generated an in vitro immunoassay system based on porcine intestinal epithelial (PIE) cells and dextran sodium sulfate (DSS) administration that could be useful for the selection and characterization of potential probiotic strains to be used in inflammatory bowel disease (IBD) patients. Our strategy was based on two fundamental pillars: on the one hand, the capacity of PIE cells to create a monolayer by attaching to neighboring cells and efficiently mount inflammatory responses and, on the other hand, the use of two probiotic bifidobacteria strains that have been characterized in terms of their immunomodulatory capacities, particularly in mouse IBD models and patients. Our results demonstrated that DSS administration can alter the epithelial barrier created in vitro by PIE cells and induce a potent inflammatory response, characterized by increases in the expression levels of several inflammatory factors including TNF-α, IL-1α, CCL4, CCL8, CCL11, CXCL5, CXCL9, CXCL10, SELL, SELE, EPCAM, VCAM, NCF2, and SAA2. In addition, we demonstrated thatBifidobacterium breveM-16V andB.longumBB536 are able to regulate the C-jun N-terminal kinase (JNK) intracellular signalling pathway, reducing the DSS-induced alterations of the in vitro epithelial barrier and differentially regulating the inflammatory response in a strain-dependent fashion. The good correlation between our in vitro findings in PIE cells and previous studies in animal models and IBD patients shows the potential value of our system to select new probiotic candidates in an efficient way.