ANTI-INFLAMMATORY FACTOR IN SACCHAROMYCES BOULARDI

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。益生菌酵母菌可以改善由多种肠道病原体引起的肠道损伤和炎症。我们发现，布氏酵母菌通过产生一种低分子量的可溶性因子SAIF(布氏酵母抗炎因子)来发挥抗炎作用，该因子可以阻断肠上皮细胞和单核细胞中MAPK和NF-kB的激活以及IL-8基因的表达。部分纯化的SAIF在艰难梭菌毒素诱导的肠炎模型中具有明显的抑制作用。我们发现，该因子对热稳定性、水溶性和蛋白酶不敏感，而对糖苷酶和芳基硫酸酯酶敏感。最近，通过离子交换，我们发现赛义夫带负电荷。简单的离子交换可使SAIF提纯100倍。P2凝胶分级过滤使SAIF组分进一步得到20倍的纯化和分离。SAIF的可能大小介于1-2个糖之间。半纯化的SAIF的质谱图显示多个糖峰，但表明该峰是由多个组分组成的。根据这些结果，进一步的纯化步骤正在进行中。抑制核因子-kB的激活是一种有吸引力的治疗靶点，可用于治疗多种人类疾病，如关节炎、哮喘、炎症性肠病以及癌症。由于布拉酵母几十年来在欧洲被广泛用作益生菌，现在在美国，SAIF可能是一种有吸引力的安全的药理制剂。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The probiotic yeast Saccharomyces boulardii ameliorates intestinal injury and inflammation caused by a wide variety of enteric pathogens. We found that Saccharomyces boulardii exerts an anti-inflammatory effect by producing a low molecular weight soluble factor SAIF (S. boulardii anti-inflammatory factor) that blocks MAPK and NF-kB activation and IL-8 gene expression mediated by both pathways in intestinal epithelial cells and monocytes. Partially purified SAIF results in a marked inhibition in the C. difficile toxin-induced enteritis model. We found that this factor is heat-stable, water-soluble and proteinase-insensitive, but glycosidases and aryl-sulfatase senstive. More recently by ion-exchange, we found that SAIF is negatively charged. Simple ion exchange can give 100-fold purification of SAIF. P2 Gel sizing filtration gave us further 20-fold purification and separation of the SAIF fraction. The possible size of SAIF is between 1-2 saccharides. Mass spectrometry of the semi-purified SAIF yielded multiple peaks indicative of saccharides, but indicated that the peak is made up of multiple components. Based on these results, further purification steps are underway. Inhibition of NF-kB activation is an attractive therapeutic target in a wide range of human diseases such as arthritis, asthma and inflammatory bowel disease as well as cancer. As Saccharomyces boulardii has been widely used as probiotics in Europe for decades and now in the U.S., SAIF could be an attractive and safe pharmacologic agent.