In vitro screening of probiotics and synbiotics according to anti-inflammatory and anti-proliferative effects
In vitro screening of probiotics and synbiotics according to anti-inflammatory and anti-proliferative effects
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DOI：
10.1016/j.ijfoodmicro.2010.09.007
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发表时间：
2010-11-15
影响因子：
5.4
通讯作者：
Rogues, Christine
中科院分区：
文献类型：
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作者：
Grimoud, Julien;Durand, Henri;Rogues, Christine
越来越多的证据表明益生菌、益生元和合生元治疗炎症性肠病 (IBD) 及其长期并发症之一——结直肠癌 (CRC) 的有效性。在这项研究中，筛选了不同菌株的益生菌乳酸菌、益生元低聚葡萄糖（GOS）或两者的合生元组合，在此类疾病的不同体外模型中筛选其抗炎和抗增殖作用。为了模拟IBD对革兰氏阴性细菌的反应，HT-29细胞通过IFNγ对脂多糖（LPS）的炎症反应敏感，从而增加了LPS生物传感器TLR4的表达，然后用益生菌、益生元和合生元进行处理。分泌的 IL-8 和活化的 NF-κ B 作为炎症生物标志物进行监测。然后选择活性菌株进行第二个炎症细胞培养模型，该模型由在 NF-κ B 诱导型启动子控制下的报告基因转染的炎症激活的转基因 Caco-2 细胞组成。报告基因表达的量化使我们能够证明一些益生菌抑制特性或在两种不同的模型中确认这些特性。通过 XTT 测定监测癌性 HT-29 细胞的增殖。只有三种益生菌菌株引起增殖减少，但缺乏重现性。二元或三元益生菌组合，无论是否由益生元 GOS 补充，均显着降低增殖，尤其是与短双歧杆菌、乳酸乳球菌和寡聚交替糖的合生组合。一个GOS。选择该组合用于以下实验。我们通过剂量范围测试证明了这种合生元的细菌和碳水化合物化合物对观察到的效果的参与。我们证明，这种增殖减少可能是由于分化表型的诱导，如肠道碱性磷酸酶（一种分化生物标志物）的上调所示，通过实时 RT-PCR 在选定的合生元处理的 HT-29 细胞中进行监测。因此，这项研究证明了益生菌发挥抗炎作用的能力，并显示了特定合生元的一些抗增殖特性。这些产品应在动物模型中进行进一步评估，以确认体外结果。 (C) 2010 Elsevier B.V. 保留所有权利。
There is emerging evidence of the efficiency of probiotic, prebiotic and synbiotic treatments in inflammatory bowel diseases (IBDs) and one of their long-term complications, colorectal cancer (CRC). In this study, various strains of probiotic lactic acid bacteria, prebiotic glucooligosaccharides (GOS) or a synbiotic combination of the two were screened for anti-inflammatory and anti-proliferative effects in different in vitro models in the context of such diseases. To mimic IBD response to Gram negative bacteria, HT-29 cells were sensitised to inflammatory response to lipopolysaccharide (LPS) by IFN gamma which increased expression of TLR4, the LPS biosensor, and were then treated by probiotics, prebiotics and synbiotics. Secreted IL-8 and activated NF-kappa B were monitored as inflammation biomarkers. A selection of active strains were then subjected to a second inflammatory cell culture model consisting of inflammatory activated transgenic Caco-2 cells transfected by a reporter gene under the control of NF-kappa B inducible promoter. Quantification of reporter gene expression allowed us to demonstrate some probiotic inhibitory properties or to confirm such characteristics in two different models. Proliferation of cancerous HT-29 cells was monitored by XTT assay. Only three probiotic strains induced a proliferation decrease, but with a lack of reproducibility. Binary or ternary probiotic associations, complemented or not by prebiotic GOS, significantly decreased proliferation, especially with a synbiotic association of Bifidobacterium breve, Lactococcus lactis and oligoalternan. a GOS. This combination was selected for the following experiments. We showed the involvement of both bacterial and carbohydrate compounds of this synbiotic in the observed effect by dose range tests. We demonstrated that this decrease in proliferation may be due to an induction of a differentiated phenotype, as shown by the up-regulation of intestinal alkaline phosphatase, a biomarker of differentiation, monitored by real-time RT-PCR in HT-29 cells treated by the selected synbiotics. Thus, this study demonstrates the ability of probiotics to exert anti-inflammatory effects and shows some anti-proliferative characteristics for a specific synbiotics. These products should be further evaluated in animal models to confirm the in vitro results. (C) 2010 Elsevier B.V. All rights reserved.