Chang Chao, Wang Kun, Zhou Sheng-Nan, Wang Xue-Dong, Wu Jin-E
College of Food Science and Engineering, Wuhan Polytechnic University, 68 South Xuefu Road, Changqing Garden, Wuhan, 430023, China.
Appl Biochem Biotechnol. 2017 May;182(1):411-427. doi: 10.1007/s12010-016-2335-x. Epub 2016 Nov 23.
The aims of our study were to evaluate the effects of Saccharomyces boulardii (S. boulardii) on deoxynivalenol (DON)-induced injury in porcine alveolar macrophage cells (PAMCs) and to explore the underlying mechanisms. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometric analysis, ELISA, qRT-PCR, and western blot were performed to assess whether S. boulardii could prevent DON-induced injury by p38 mitogen-activated protein kinase (p38 MAPK) signal pathway. The results showed that pretreatment with 8 μM DON could decrease the viability of PAMC and significantly increase the apoptosis rate of PAMC, whereas S. boulardii could rescue apoptotic PAMC cells induced by DON. Further experiments revealed that S. boulardii effectively reversed DON-induced cytotoxicity via downregulating the expression of TNF-α, IL-6, and IL-lβ. In addition, S. boulardii significantly alleviated DON-induced phosphorylation and mRNA expression of p38 and further increased the expression of apoptosis regulation genes Bcl-xl and Bcl-2 and inhibited the activation of Bax. Our results suggest that S. boulardii could suppress DON-induced p38 MAPK pathway activation and reduce the expression of downstream inflammatory cytokines, as well as promote the expression of anti-apoptotic genes to inhibit apoptosis induced by DON in PAMC.
我们研究的目的是评估布拉氏酵母菌（S. boulardii）对猪肺泡巨噬细胞（PAMCs）中脱氧雪腐镰刀菌烯醇（DON）诱导损伤的影响，并探索其潜在机制。采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法、流式细胞术分析、酶联免疫吸附测定（ELISA）、定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法，以评估布拉氏酵母菌是否能通过p38丝裂原活化蛋白激酶（p38 MAPK）信号通路预防DON诱导的损伤。结果表明，用8 μM DON预处理可降低PAMC的活力，并显著提高PAMC的凋亡率，而布拉氏酵母菌可挽救由DON诱导的凋亡PAMC细胞。进一步实验表明，布拉氏酵母菌通过下调肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和白细胞介素-1β（IL-1β）的表达有效逆转了DON诱导的细胞毒性。此外，布拉氏酵母菌显著减轻了DON诱导的p38磷酸化和mRNA表达，并进一步增加了凋亡调控基因Bcl-xl和Bcl-2的表达，抑制了Bax的激活。我们的结果表明，布拉氏酵母菌可抑制DON诱导的p38 MAPK信号通路激活，降低下游炎性细胞因子的表达，并促进抗凋亡基因的表达，以抑制DON诱导的PAMC细胞凋亡。
