酿酒酵母细胞壁诱导体外培养绵羊瘤胃上皮细胞表达SBD-1的研究

doi:10.11843/j.issn.0366-6964.2018.05.007

摘要/Abstract

摘要：

旨在探究酿酒酵母细胞壁对原代培养绵羊瘤胃上皮细胞β-防御素-1（Sheep beta-defensin-1，SBD-1）表达的影响。本试验首先提取酿酒酵母细胞壁，并进行鉴定；再用酿酒酵母细胞壁（0、25、50、100、200、400 μg·mL^-1）刺激健康绵羊的原代瘤胃上皮细胞（0、2、4、8、12、24 h）后，进行RT-qPCR和ELISA试验，确定诱导SBD-1表达的最佳浓度和时间。结果表明：1）本试验提取的酿酒酵母细胞壁成分较纯，可用于后续试验；2）体外培养绵羊瘤胃上皮细胞呈鹅卵石铺路状，符合试验要求；3）使用200 μg·mL^-1的酿酒酵母细胞壁去刺激瘤胃上皮细胞12 h，SBD-1 mRNA和蛋白表达量最大，与其他组相比均差异极显著（P<0.001）。综上表明，酿酒酵母细胞壁能够促进绵羊瘤胃上皮细胞SBD-1的表达，且酵母细胞壁最佳浓度和刺激时间分别为200 μg·mL^-1和12 h。

关键词: 酿酒酵母, 细胞壁, 绵羊, 瘤胃上皮细胞, β-防御素-1

Abstract:

In present study, we aimed to explore the effect of the cell wall of Saccharomyces cerevisiae (S. cerevisiae) on the expression of sheep beta defensin-1 (SBD-1) in primary culture of sheep's ruminal epithelial cells (SRECs). Firstly, the cell wall of S.cerevisiae was extracted and subjected to quality test. Then, SRECs were cultured and stimulated with cell wall of S. cerevisiae (0, 25, 50, 100, 200, 400 μg·mL^-1) for different times (0, 2, 4, 8, 12, 24 h). RT-PCR and ELISA were performed to grope the optimal concentration and time for SBD-1 expression. The results showed that:1) The cell wall of S. cerevisiae extracted was pure and suitable for the subsequent experiments; 2) SRECs were in vitro cultured successfully (cobblestone pavement), that meet the experimental requirement; 3) Compared with other concentration groups, the expression of SBD-1 mRNA and protein reached the highest when the cell wall of S. cerevisiae of 200 μg·mL^-1 was used to stimulate SRECs for 12 h (P<0.001). The results indicated that the cell wall of S. cerevisiae could improve the expression of SBD-1 in SRECs, and optimal condition was 200 μg·mL^-1for 12 h.

Key words: S. cerevisiae, cell wall, sheep, ruminal epithelial cells, β-defensin-1

中图分类号:

S826.2

田巧珍, 金鑫, 张曼, 张召议, 王云鹤, 杨银凤. 酿酒酵母细胞壁诱导体外培养绵羊瘤胃上皮细胞表达SBD-1的研究[J]. 畜牧兽医学报, 2018, 49(5): 927-934.

TIAN Qiao-zhen, JIN Xin, ZHANG Man, ZHANG Zhao-yi, WANG Yun-he, YANG Yin-feng. Effects of Cell Wall of Saccharomyces cerevisiae on the Expression of SBD-1 in Cultured Ruminal Epithelial Cells of Sheep[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(5): 927-934.

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