酿酒酵母菌及其灭活菌对绵羊瘤胃外植体β-防御素-1(SBD-1)表达的影响  

Effects of Saccharomyces Cerevisiae and Its Inactivated Bacteria on the Expression of β-defensin-1 in Ruminal Explants of Sheep

作　　者：王云鹤[1,2] 金鑫[1,2] 张曼[1,2] 魏方 温婧怡 赵霏霏 杨银凤[1,2] WANG Yunhe;JIN Xin;ZHANG Man;WEI Fang;WEN Jingyi;ZHAO Feifei;YANG Yinfeng(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease ofMinistry of Agriculture,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学兽医学院,呼和浩特010018 [2]农业部动物疾病临床诊疗技术重点实验室,呼和浩特010018

出　　处：《畜牧兽医学报》2019年第3期581-591,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31560682)

摘　　要：旨在探索益生性酿酒酵母菌及其灭活菌对绵羊瘤胃外植体内β-防御素-1(sheep beta-defensin-1,SBD-1)表达的影响。本研究建立了绵羊瘤胃外植体培养方法,将不同浓度(10~4、10~5、10~6、10~7、10~8、10~9 CFU·mL^(-1))的酿酒酵母菌及其灭活菌与外植体共培养24 h后,通过qPCR和ELISA方法检测瘤胃外植体内SBD-1 mRNA和蛋白的表达变化,以分别确定活菌及其灭活菌诱导SBD-1表达最高的菌液浓度;然后用该浓度的活菌及其灭活菌对瘤胃外植体进行不同时间(2、4、8、12、16、20、24 h)的刺激,同样用qPCR和ELISA方法检测瘤胃外植体内SBD-1 mRNA及蛋白的表达变化,从而筛选出活菌及其灭活菌诱导SBD-1表达的最佳时间。结果表明,益生性酿酒酵母菌及其灭活菌均能显著促进绵羊瘤胃外植体SBD-1的表达(P<0.05)。当酿酒酵母菌浓度为10~7 CFU·mL^(-1)、灭活菌浓度为10~8 CFU·mL^(-1)分别诱导绵羊瘤胃外植体16 h时,SBD-1表达量分别达到最大,且二者与对照相比均呈极显著差异(P<0.01)。在最佳诱导条件下对比二者的诱导效果,酿酒酵母菌活菌高于其灭活菌。因此,益生性酿酒酵母菌及其灭活菌均能促进绵羊瘤胃外植体内SBD-1的表达,且活菌浓度为10~7 CFU·mL^(-1)、灭活菌浓度为10~8 CFU·mL^(-1)分别诱导16 h时,绵羊瘤胃外植体内的SBD-1的表达量分别达到最大,并且益生性酿酒酵母菌的诱导效果高于其灭活菌。The aim of this study was to explore the effects of probiotic Saccharomyces cerevisiae ( S.c ) and inactivated S.c on the expression of sheep-defensin-1 (SBD-1) in rumen explants of sheep. After different concentrations (10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 CFU·mL -1 ) of S.c and inactivated S.c were respectively co-cultured with explants for 24 h, the expression of SBD-1 mRNA and protein in rumen explants were detected by qPCR and ELISA to determine the optimal concentration for the induction of SBD-1 expression by S.c and inactivated S.c . Then, the sheep rumen explants were stimulated with the optimal concentration of S.c and inactivated S.c for 2, 4, 8, 12, 16, 20, 24 h, respectively, and the SBD-1 mRNA and protein were also detected by qPCR and ELISA, so as to screen out the optimal time for the induction of SBD-1 expression by S.c and inactivated S.c . The results showed that S.c and inactivated S.c could significantly promote the expression of SBD-1 in rumen explants ( P <0.05). When the rumen explants of sheep were stimulated by the concentration of 10 7 CFU·mL -1 for S.c and 10 8 CFU·mL -1 for inactivated S.c for 16 h, respectively, the expression level of SBD-1 were the highest, and which were significantly different from the control ( P <0.01). Comparing the induction effects between S.c and inactivated S.c under optimal induction conditions, the S.c were better. Therefore, S.c and inactivated S.c could promote the SBD-1 expression in rumen explants of sheep, and the expression of SBD-1 reached the maximum when the rumen explants of sheep were stimulated by the concentration of 10 7 CFU·mL -1 for S.c and 10 8 CFU·mL -1 for inactivated S.c for 16 h. The induction effect of S.c is better than that of inactivated S.c .

关 键 词：酿酒酵母菌 β-防御素-1 绵羊瘤胃外植体 qPCR ELISA 

分 类 号：S826[农业科学—畜牧学]