枯草芽孢杆菌对绵羊瘤胃上皮细胞β-防御素-1表达的影响

Effects of Bacillus subtilis on the Expression of SBD-1 in Cultured Ruminal Epithelial Cells of Sheep

选用益生枯草芽孢杆菌研究其对体外培养的绵羊瘤胃上皮细胞β-防御素表达的调节作用。首先,在体外成功培养绵羊瘤胃上皮细胞,然后用枯草芽孢杆菌对体外培养的绵羊瘤胃上皮细胞进行不同浓度、不同时间的刺激,利用荧光定量PCR技术(Real-time fluorescence quantitative PCR,RT-PCR)从mRNA水平检测刺激后上皮细胞中绵羊β-防御素-1(Sheep beta-defensin-1,SBD-1)基因表达水平的差异。结果表明:当菌液浓度为1010 cfu·mL-1刺激上皮细胞8h后,SBD-1的表达量达到最高;不同的菌液浓度诱导下SBD-1的表达量均有显著增加,109、1010、1011 cfu·mL-1菌液浓度刺激下,SBD-1的表达量与空白相比差异极显著(P<0.01)。结果表明,枯草芽孢杆菌能够诱导绵羊瘤胃上皮细胞内SBD-1基因的表达。

In this study,Bacillus subtilis was selected to investigate its effect on theβ-defensin expression of ovine ruminal epithelial cells cultured in vitro.Firstly,the ovine ruminal epithelial cells were cultured in vitro,then the epithelial cells were stimulated by Bacillus subtilis with different concentrations at different times,the expression of SBD-1mRNA was determined by realtime fluorescence quantitative PCR(RT-PCR).The results indicated that the expression peak of SBD-1mRNA was after 8htreated by Bacillus subtilis with 1010 cfu·mL-1.The expression of SBD-1was increased significantly after treated by Bacillus subtilis with different concentrations,and the expression at 109,1010,1011 cfu·mL-1showed stronger induction response than the blank(P<0.01).This study reveals that Bacillussubtiliscan induce the expression of SBD-1in ovine ruminal epithelial cells.