微生物环境及其相应的分泌代谢产物谱是肠上皮细胞功能的重要调节因子,影响整个有机体。猪空肠上皮细胞系(IPEC-J2)是在不同细胞培养模型中肠细胞分化的体外模型。氧气供应的改善似乎是气液界面培养中分化的主要原因,但这一点尚未得到最终澄清。在这种情况下,细胞的营养及其对新陈代谢的影响也至关重要。由于短链脂肪酸(SCFA)在某些疾病(如多发性硬化症和其他炎性疾病)中的临床相关性,近年来对短链脂肪酸(SCFA)的兴趣稳步增长。但对猪中的FFAR2和FFAR3(游离脂肪酸受体2和3)知之甚少。我们要解决的问题:1。关于FFAR2和FFAR3在susscrofa2体内和体外的分布。是否有丙酸的影响,葡萄糖含量和培养对肠上皮细胞代谢的影响?通过对猪肠段空肠的冷冻切片进行免疫染色,研究了FFAR2和FFAR3在体内的形态学分析,回肠和结肠.两种受体均沿肠表达,并在肌层和粘膜肌层的平滑肌细胞中发现。此外,在空肠中也观察到肠神经系统中FFAR2的高表达和FFAR3的低表达,susscrofa的回肠和结肠。此外,对血管内的FFAR2和FFAR3进行了调查。FFAR3在静脉和淋巴管的内皮细胞上显示出强表达,但在动脉上未检测到。此外,我们第一次展示,FFAR2和FFAR3在IPEC-J2细胞RNA和蛋白质水平上,以及共聚焦显微镜。此外,在IPEC-J2细胞中,ENO1和NDUFA4作为2个重要基因在RNA水平上进行了研究,在新陈代谢中起着至关重要的作用。这里,在模型动物susscrofa以及猪细胞系IPEC-J2中检测到NDUFA4。用海马分析仪测试丙酸和/或葡萄糖和/或培养方法对细胞代谢的潜在影响。这里,在SMC中观察到的ECAR明显高于OCR。总之,我们能够证明,培养系统似乎比培养基成分或细胞营养有更大的影响。然而,这可以通过孵育时间或不同SCFA的组合来调节。
The microbiological environment and their corresponding secreted metabolite spectrum are an essential modulator of the enterocyte function, effecting the whole organism. Intestinal porcine jejunal epithelial cell line (IPEC-J2) is an established in vitro model for differentiation of
enterocytes in different cell culture models. An improved oxygen supply seems to be the main reason for differentiation in an air-liquid-interface culture, but this has not yet been conclusively clarified. In this context, the nutrition of the cell and its influence on the metabolism is also of crucial importance. The interest in short-chain fatty acids (SCFAs) has grown steadily in recent years due to their clinical relevance in certain diseases such as multiple sclerosis and other inflammatory diseases, but not much is known of FFAR2 and FFAR3 (free fatty acid receptor 2 and 3) in pigs. We want to address the questions: 1. about the distribution of FFAR2 and FFAR3 in vivo and in vitro in sus scrofa 2. whether there is an influence of propionic acid, glucose content and cultivation on metabolism of
enterocytes? The morphological analysis of FFAR2 and FFAR3 in vivo was investigated through immunostaining of frozen sections of the porcine gut segments jejunum, ileum and colon. Both receptors are expressed along the gut and were found in the smooth muscle cells of the tunica muscularis and lamina muscularis mucosae. Furthermore, a high expression of FFAR2 and a low expression of FFAR3 in the enteric nerve system was also observed in jejunum, ileum and colon of sus scrofa. In addition, FFAR2 and FFAR3 within the vessels was investigated. FFAR3 showed a strong expression on endothelial cells of veins and lymphatic vessels but was not detectable on arteries. Furthermore, we demonstrate for the first time, FFAR2 and FFAR3 in IPEC-J2 cells on RNA- and protein level, as well as with confocal microscopy. In addition, ENO1 and NDUFA4 were investigated on RNA-level in IPEC-J2 cells as 2 important genes, which play an essential role in metabolism. Here, NDUFA4 is detected in the model animal sus scrofa as well as in the porcine cell line IPEC-J2. A potential impact of propionic acid and/or glucose and/or cultivation method on the metabolism of the cells was tested with the Seahorse analyzer. Here, a significant higher ECAR was observed in the SMC than in the OCR. In summary, we were able to show that the cultivation system appears to have a greater influence than the medium composition or nutrition of the cells. However, this can be modulated by incubation time or combination of different SCFAs.