酿酒酵母β-葡聚糖对绵羊瘤胃上皮细胞β-防御素-1表达的影响

doi:10.11843/j.issn.0366-6964.2018.11.013

Abstract

Abstract:

The aim of this study was to explore the effect of Saccharomyces cerevisiae β-glucan on the β-defensin-1(SBD-1) expression in cultured ruminal epithelial cells(RECs) of ovine. Firstly, the ovine RECs culture system was established as an in vitro experimental model, the RECs were stimulated with various concentrations (0, 5, 10, 20, 50 and 100 μg·mL^-1) of β-glucan for 8 h, and the expression level of SBD-1 mRNA and protein in RECs were detected by qPCR and ELISA to determine the concentration of β-glucan that induced the highest expression of SBD-1. Then, RECs were stimulated with the optimal stimulating concentration of β-glucan for 0, 2, 4, 8, 12 and 24 h, respectively and the expression of RECs SBD-1 were also detected by qPCR and ELISA, so as to determine the optimal stimulation time for β-glucan-induced SBD-1 expression. The qPCR and ELISA results showed that after β-glucan (5-100 μg·mL^-1) stimulated RECs for 8 h, the expression of SBD-1 mRNA and protein were increased and then decreased following the increase of concentrations, and when the concentration of β-glucan was 10 μg·mL^-1, the expression level of SBD-1 mRNA and protein were significantly higher than the control group (P<0.01). When RECs were stimulated with 10 μg·mL^-1 β-glucan for 0-24 h, qPCR results showed that SBD-1 mRNA expression was the highest at 2 h (P<0.01), and then downward; The results of ELISA test showed that SBD-1 protein secretion reached the highest level at 4 h (P<0.01).The results of MTT test showed that 100 μg·mL^-1 β-glucan significantly affect the sheep RECs viability (P<0.05). The results of this study indicated that β-glucan could improve the SBD-1 expression in RECs of ovine. And the level of SBD-1 mRNA and protein expression was the highest when the RECs were separately stimulated for 2 and 4 h with the 10 μg·mL^-1 β-glucan.

Key words: β-defensin-1, ruminal epithelial cells, β-glucan, qPCR, ELISA, MTT

CLC Number:

S826.2

ZHANG Man, JIN Xin, WANG Yun-he, WEI Fang, WEN Jing-yi, LI Zheng-yi, YANG Yin-feng. Effects of Saccharomyces cerevisiae β-glucan on the Expression of SBD-1 in Ovine Ruminal Epithelial Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(11): 2416-2424.

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Effects of Saccharomyces cerevisiae β-glucan on the Expression of SBD-1 in Ovine Ruminal Epithelial Cells

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