酿酒酵母菌诱导SBD-1 mRNA表达的研究

Study on the expression of SBD-1 mRNA induced by Saccharomyces cerevisiae

为了探索益生性酿酒酵母菌对绵羊瘤胃上皮细胞β-防御素-1(Sheep Beta-Defensin-1,SBD-1)表达的调节作用。建立绵羊瘤胃上皮细胞培养体系作为体外试验模型,用不同浓度酿酒酵母菌对体外培养的绵羊瘤胃上皮细胞刺激2 h后分别进行不同时间的诱导培养,然后利用实时荧光定量PCR技术(RT-q PCR)研究接受刺激后瘤胃上皮细胞中SBD-1 mRNA表达的差异。结果表明,在各时间组内,SBD-1 mRNA的表达量随着菌液浓度的增加均呈先升高后降低的趋势,当菌液浓度为5.2×107CFU/m L时表达量均达到最大,并与对照组和其他浓度组相比差异极显著(P﹤0.01)。当菌液浓度一定时,SBD-1 mRNA的表达量先是随着诱导培养时间的延长而呈上升趋势,诱导培养时间为12 h时SBD-1 mRNA表达量达到峰值,之后呈下降趋势。因此,当浓度为5.2×107CFU/m L的菌液诱导瘤胃上皮细胞12 h时,SBD-1基因的表达量达到最高,且与此浓度下其他时间组的表达量相比差异极显著(P﹤0.01)。本试验结果表明,酿酒酵母菌能够诱导绵羊瘤胃上皮细胞SBD-1的表达,且浓度为5.2×107CFU/m L的菌液诱导瘤胃上皮细胞12 h时,SBD-1的表达量达到最高。

In this study, we investigated the effect of probiotic Saccharomyces cerevisiae on the β - defensin-1 （ Sheep Beta-De- fensin-1, SBD-1 ） expression in cultured ruminal epithelial cells of sheep. The ruminal epithelium cells（RECs） of sheep were cul- tured successfully in vitro, and the RECs were stimulated 2 h with different concentrations of Saccharomyces cerevisiae, and then for different times. Then the expression of SBD-1 mRNA was analyzed by real-time fluorescence quantitative PCR（RT-qPCR）. The re- sults showed that the expression of SBD-1 mRNA in each time group were increased and then decreased following the increase of Sac- charomyces eerevisiae concentrations. The expression of SBD - 1 mRNA was highest and significantly higher than the control group and the other concentration groups （P 〈 O. O1 ）. When the cells were stimulated with the concentration of 5.2 x lO7 CFU - mL-1. When the concentration was constant, the expression of SBD - 1 mRNA was increased with the extension of stimulating time, and the expression of SBD-1 mRNA was the highest at 12 h. Therefore, the expression of SBD-1 was the highest when the concentration of Saccharomyces cerevisiae was at 5.2 × 107CFU mL-1 and the stimulating was 12 h stimulating, compared with the other time groups at this concentration （P 〈0.01 ）. These findings indicate that Saccharomyces cerevisiae can induce the expression of SBD-1 in ruminal epithelial cells of sheep. And the level of SBD-1 expression is highest when the rumen epithelial cells are stimulated with the concentration of 5.2× 107CFU mL-lfor 12 h.