【摘要】 为了探究来源于枯草芽孢杆菌细胞壁的肽聚糖对绵羊瘤胃上皮细胞(ORECs)中β-防御素-1(SBD-1)表达的影响，本试验使用30μg/mL肽聚糖刺激ORECs不同时间(2、4、6、8、10和12 h),采用荧光定量PCR(qPCR)和酶联免疫吸附试验(ELISA)方法分别检测SBD-1的mRNA和蛋白表达量，确定最佳刺激时间；再使用不同浓度的肽聚糖(0、10、20、30、40和50μg/mL)刺激ORECs 2 h,采用qPCR和ELISA方法检测SBD-1的mRNA和蛋白表达量，确定最佳刺激浓度。结果显示：(1)使用30μg/mL肽聚糖刺激ORECs不同时间时，SBD-1的mRNA和蛋白表达量随着时间的增加呈先下降后升高的趋势，2 h时表达量最多并极显著高于对照组(P<0.001)。(2)使用不同浓度肽聚糖刺激ORECs 2 h时，SBD-1的mRNA和蛋白表达量随着肽聚糖浓度的升高呈现先升高后降低的趋势，在20μg/mL时表达量最多并极显著高于对照组(P<0.001)。(3)不同时间刺激试验和不同浓度刺激试验均不会对ORECs产生细胞毒性作用。结果表明，来源于枯草芽孢杆菌的肽聚糖能够诱导ORECs表达SBD-1,最佳刺激条件为20μg/mL枯草芽孢杆菌肽聚糖刺激ORECs 2 h。更多还原

【Abstract】 To explore the influence of peptidoglycan derived from Bacillus subtilis cell walls on the expression of β-defensin-1(SBD-1) in ovine rumen epithelial cells(ORECs), this study stimulated ORECs with 30 μg/mL peptidoglycan at different durations(2, 4, 6, 8, 10, and 12 h). The mRNA and protein expression levels of SBD-1 were detected using quantitative fluorescence PCR(qPCR) and enzyme-linked immunosorbent assay(ELISA), respectively, to determine the optimal stimulation time. Subsequently, ORECs were stimulated with different concentrations of peptidoglycan(0, 10, 20, 30, 40, and 50 μg/mL) for 2 h, and the mRNA and protein expression levels of SBD-1 were detected using qPCR and ELISA to determine the optimal stimulation concentration. The results showed that:(1) When ORECs were stimulated with 30 μg/mL peptidoglycan for different durations, the mRNA and protein expression levels of SBD-1 initially decreased and then increased with time, with the highest expression at 2 h, significantly higher than the control group(P<0.001).(2) When ORECs were stimulated with different concentrations of peptidoglycan for 2 h, the mRNA and protein expression levels of SBD-1 increased initially and then decreased with the concentration of peptidoglycan, reaching the highest expression at 20 μg/mL, significantly higher than the control group(P<0.001).(3) Stimulation experiments at different times and different concentrations did not induce cytotoxicity in ORECs. The results indicate that peptidoglycan derived from Bacillus subtilis can induce the expression of SBD-1 in ORECs,with the optimal stimulation condition being 20 μg/m L peptidoglycan from Bacillus subtilis for 2 h.更多还原