Study on Signaling Pathway of Saccharomyces cerevisiae Mannan-induced SBD-1 Expression in Rumen Epithelial Cells of Sheep (Ovis aries)
JIN Xin, ZHANG Man, YANG Yin-Feng*
College of Veterinary, Inner Mongolia Agricultural University/Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease, Ministry of Agriculture, Hohhot 010018, China
Abstract：β-defensins are the endogenous antibacterial peptide of the organism. Saccharomyces cerevisiae mannan can induce ovine Ruminal epithelial cells (RECs) β-defensin-1 (SBD-1) expression. However, its mechanism of inducing SBD-1 expression is unclear, which limits the development and utilization of mannan preparations. In order to explore whether mitogen-activated protein kinase (MAPK) (p38, ERK1/2, JNK) and Nuclear factor κB (NF-κB) are involved in the expression of SBD-1 in ovine RECs induced by mannan. Firstly, the effects of mannan on the expression of p38, ERK1/2, JNK and NF-κB were examined by qPCR and Western blot. Then the phosphorylation levels of p38, ERK1/2, JNK, IκB and p65 were detected by Western blot after stimulation of RECs with mannan for 5, 15, 30, 45 and 60 min, respectively. Finally, qPCR and ELISA were used to detect the effects of specific inhibitors of p38, ERK1/2, JNK and NF-κB on mannan-induced SBD-1 expression to determine the involvement of p38, ERK1/2, JNK and NF-κB pathways in the expression of SBD-1 induced by mannan. The results showed that mannan stimulation significantly increased the mRNA levels of p38, ERK1/2, JNK and NF-κB in RECs and the phosphorylation levels of p38, JNK, ERK1/2, IκB and p65 (P<0.01). And mannan stimulated RECs different times and activated phosphorylation of p38, ERK1/2, JNK, IκB and p65. Moreover, treatment with p38, ERK1/2, JNK and NF-κB inhibitors significantly decreased the expression of SBD-1 induced by mannan (P<0.01), and the inhibitory effect of p38 inhibitor on SBD-1 was the most obvious. These results indicate that the S. cerevisiae mannan-induced SBD-1 expression in RECs is mediated by MAPK and NF-κB signaling pathways, and the p38 signaling pathway may play a major role. This study explains the function of mannan to promote immunity from the mechanism of mannan-induced defensin expression, and provides a theoretical basis for better development and utilization of mannan preparations.
金鑫, 张曼, 杨银凤. 酿酒酵母甘露聚糖诱导绵羊瘤胃上皮细胞SBD-1表达的信号通路研究[J]. 农业生物技术学报, 2019, 27(6): 1034-1041.
JIN Xin ZHANG Man YANG Yin-Feng. Study on Signaling Pathway of Saccharomyces cerevisiae Mannan-induced SBD-1 Expression in Rumen Epithelial Cells of Sheep (Ovis aries). 农业生物技术学报, 2019, 27(6): 1034-1041.
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