【摘要】 以体外培养的绵羊瘤胃上皮细胞（ovine rumen epithelial cells, ORECs）为试验模型,旨在探究灭活枯草芽孢杆菌对ORECs绵羊β-防御素-1（sheepβ-defesin-1,SBD-1）表达的影响。首先用不同浓度的灭活枯草芽孢杆菌(10⁷、10⁸、10⁹、10¹⁰、10¹¹ CFU/mL)诱导ORECs 8 h,通过荧光定量PCR（qPCR）和ELISA的方法检测ORECs SBD-1表达量在mRNA水平和蛋白水平的变化,确定灭活枯草芽孢杆菌诱导ORECs SBD-1表达最高的菌液浓度为最佳刺激浓度。以该浓度的灭活枯草芽孢杆菌诱导ORECS不同时间（2 h、4 h、8 h、12 h）,通过qPCR和ELISA的方法检测ORECs SBD-1表达量在mRNA水平和蛋白水平的变化,从而筛选出诱导SBD-1表达的最佳时间。结果表明,10⁸～10¹¹ CFU/mL的灭活枯草芽孢杆菌可以显著的促进ORECs SBD-1的表达,其中灭活枯草芽孢杆菌的浓度为10¹⁰ CFU/mL时诱导ORECs 8h时SBD-1 mRNA表达量最高,与对照组相比差异极显著（P<0.01）;SBD-1蛋白表达量以10¹⁰ CFU/mL灭活枯草芽孢杆菌诱导ORECs 12 h时达到最大,与对照组相比差异极显著（P<0.01）。结果证明了热灭活后的枯草芽孢杆菌仍可以诱导SBD-1表达,其诱导SBD-1表达的有效成分并未被热灭活,为后续确定枯草芽孢杆菌诱导SBD-1表达的有效成分的研究提供了理论基础。更多还原

【Abstract】 In this study, ovine rumen epithelial cells（ORECs） cultured in vitro were used as an experimental model to investigate the effect of inactivated Bacillus subtilis on the expression of sheep β-defesin-1（SBD-1） in ORECs.Firstly, ORECs were stimulated with different concentrations of inactivated Bacillus subtilis(10⁷-10¹¹ CFU/mL) for 8 h.The changes of ORECs SBD-1 expression levels at mRNA and protein levels were detected by qPCR and ELISA methods, and the optimal stimulation concentration was determined to be the bacterial liquid concentration with the highest expression of ORECs SBD-1 induced by inactivated Bacillus subtilis.The expression level of SBD-1 in ORECs was detected by qPCR and ELISA,and the best time to stimulate the expression of SBD-1 was selected by using the inactivated Bacillus subtilis at this concentration for different time（2 h, 4 h, 8 h, 12 h）.The results showed that 10⁸-10¹¹ CFU/mL of inactivated Bacillus subtilis could significantly promote the expression of SBD-1 in ORECs, and the highest expression of SBD-1 mRNA was observed when the concentration of inactivated Bacillus subtilis was 10¹⁰ CFU/mL for 8 h of ORECs induction, and the difference was very significant compared with the control group（P<0.01）.The expression of SBD-1 protein reached the maximum after 12 h induction of ORECs by 10¹⁰ CFU/mL Bacillus subtilis inactivation, and the difference was very significant compared with the control group（P<0.01）.This experiment proved that Bacillus subtilis could still induce the expression of SBD-1 after being inactivated by heat, and the active components inducing the expression of SBD-1 were not inactivated by heat, which provided a theoretical basis for the subsequent study on the effective components inducing the expression of SBD-1 by Bacillus subtilis.更多还原