枯草芽孢杆菌肽聚糖对绵羊瘤胃上皮细胞β-防御素-1(SBD-1)表达的影响  

Effects of Bacillus subtilis Peptidoglycan on Expression ofβ-defensin-1(SBD-1)in Ovine Rumen Epithelial Cells

作　　者：齐盟 辛雅明 白绥明 胡亮 杨银凤[1,2] QI Meng;XIN Ya-ming;BAI Sui-ming;HU Liang;YANG Yin-feng(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;Key Laboratory of Basic Veterinary Medicine of Inner Mongolia Autonomous Region,Hohhot 010018,China;Erdos Animal Disease Prevention and Control Center of Inner Mongolia,Erdos 017000,China;Bayannur Institute of Agricultural and Animal Husbandry,Bayannur 015000,China)

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]内蒙古自治区基础兽医学重点试验室,内蒙古呼和浩特010018 [3]内蒙古鄂尔多斯市动物疫病预防和控制中心,内蒙古鄂尔多斯017000 [4]巴彦淖尔市农牧业科学研究所,内蒙古巴彦淖尔015000

出　　处：《中国兽医杂志》2023年第11期52-58,共7页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金项目(32060780);内蒙古自治区教育厅项目(NJZZ21015)。

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。To explore the influence of peptidoglycan derived from Bacillus subtilis cell walls on the expression ofβ-defensin-1(SBD-1)in ovine rumen epithelial cells(ORECs),this study stimulated ORECs with 30μg/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 50μg/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 30μg/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 20μg/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 20μg/mL peptidoglycan from Bacillus subtilis for 2 h.

关 键 词：枯草芽孢杆菌肽聚糖 绵羊瘤胃上皮细胞(ORECs) β-防御素-1(SBD-1) 荧光定量PCR(qPCR) 酶联免疫吸附试验(ELISA) CCK8 

分 类 号：S858.26[农业科学—临床兽医学]