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

doi:10.11843/j.issn.0366-6964.2019.03.013

摘要/Abstract

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（10⁴、10⁵、10⁶、10⁷、10⁸、10⁹ 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10⁷ CFU·mL^-1、灭活菌浓度为10⁸ 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10⁷ CFU·mL^-1、灭活菌浓度为10⁸ 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。

Abstract:

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 (10⁴, 10⁵, 10⁶, 10⁷, 10⁸, 10⁹ 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 10⁷ CFU·mL^-1 for S.c and 10⁸ 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 10⁷ CFU·mL^-1 for S.c and 10⁸ CFU·mL^-1 for inactivated S.c for 16 h. The induction effect of S.c is better than that of inactivated S.c.

中图分类号:

S826

王云鹤, 金鑫, 张曼, 魏方, 温婧怡, 赵霏霏, 杨银凤. 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[J]. 畜牧兽医学报, 2019, 50(3): 581-591.

WANG Yunhe, JIN Xin, ZHANG Man, WEI Fang, WEN Jingyi, ZHAO Feifei, YANG Yinfeng. Effects of Saccharomyces Cerevisiae and Its Inactivated Bacteria on the Expression of β-defensin-1 in Ruminal Explants of Sheep[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(3): 581-591.

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