Song Sooyeon, Lee Sei-Jung, Park Dong-June, Oh Sejong, Lim Kye-Taek
Department of Food Science, Cornell University, 411 Tower Road, Ithaca, NY 14853.
Department of Veterinary Physiology, College of Veterinary Medicine, Research Institute for Veterinary Science, and BK21 PLUS Creative Veterinary Research Center, Seoul National University, Seoul 08826, South Korea.
J Dairy Sci. 2016 Dec;99(12):9372-9382. doi: 10.3168/jds.2016-11809. Epub 2016 Oct 13.
Recently, interest in the beneficial role of probiotics in the protection and management of allergic diseases caused by immune disorders has been increasing. This study investigated the inhibitory effect of Lactobacillus plantarum L67 on induced allergic inflammatory response in bisphenol A-treated rat basophilic leukemia 2H3 (RBL-2H3) cells and mouse splenocytes. We also evaluated the applicability of L. plantarum L67 as a yogurt starter culture. We measured the ability of Lactobacillus strains to induce the production of IL-12 and IFN- γ in cultured splenocytes by ELISA. Bisphenol A (50μM)-treated RBL-2H3 cells were cotreated with a glycoprotein (18kDa) isolated from L. plantarum L67 (5-100µg/mL) for 30min. We measured the expression of mitogen-activated protein kinase (ERK and p38), AP-1 (c-Fos and c-Jun), T-bet, and GATA-binding protein 3 (GATA-3) using Western blotting to examine the differentiation of T helper cells. Furthermore, we evaluated the gene expression of IL-1β, IL-6, and IL-10 using real-time quantitative PCR. Finally, we evaluated the applicability of L. plantarum L67 as a yogurt starter by measuring pH, enumeration of bacteria, and sensory scores. Our results showed that L67 protein inhibited the phosphorylation of ERK and p38 mitogen-activated protein kinase through the transcriptional activation of AP-1 in bisphenol A-treated RBL-2H3 cells. During differentiation of T helper cells, the expression of transcription factor GATA-3 was significantly suppressed by L67 protein (100µg/mL) treatment, whereas expression of transcription factor T-bet was increased. In addition, the L67 protein significantly attenuated the expression of T helper 2-linked cytokines IL-1β, IL-6, and IL-10. These results indicate that L. plantarum L67, made available as yogurt starters and dietary supplements, has the potential to prevent allergy-related immune disorders.
最近，益生菌在免疫紊乱引起的过敏性疾病的保护和管理中的有益作用受到越来越多的关注。本研究调查了植物乳杆菌L67对双酚A处理的大鼠嗜碱性白血病2H3（RBL-2H3）细胞和小鼠脾细胞诱导的过敏性炎症反应的抑制作用。我们还评估了植物乳杆菌L67作为酸奶发酵剂的适用性。我们通过ELISA测定了乳酸菌菌株在培养的脾细胞中诱导IL-12和IFN-γ产生的能力。用从植物乳杆菌L67中分离的糖蛋白（18kDa）（5-100µg/mL）与双酚A（50μM）处理的RBL-2H3细胞共处理30分钟。我们使用蛋白质印迹法测量丝裂原活化蛋白激酶（ERK和p38）、AP-1（c-Fos和c-Jun）、T-bet和GATA结合蛋白3（GATA-3）的表达，以检查辅助性T细胞的分化。此外，我们使用实时定量PCR评估IL-1β、IL-6和IL-10的基因表达。最后，我们通过测量pH值、细菌计数和感官评分来评估植物乳杆菌L67作为酸奶发酵剂的适用性。我们的结果表明，L67蛋白通过AP-1在双酚A处理的RBL-2H3细胞中的转录激活来抑制ERK和p38丝裂原活化蛋白激酶的磷酸化。在辅助性T细胞分化过程中，L67蛋白（100µg/mL）处理显著抑制了转录因子GATA-3的表达，而转录因子T-bet的表达增加。此外，L67蛋白显著减弱了辅助性T细胞2相关细胞因子IL-1β、IL-6和IL-10的表达。这些结果表明，作为酸奶发酵剂和膳食补充剂提供的植物乳杆菌L67有预防过敏相关免疫紊乱的潜力。
