乳酸杆菌诱导绵羊瘤胃上皮细胞内SBD-1的表达  

The expression of SBD-1 in the rumen epithelium cells of sheep induced by lactobacillus

作　　者：范燕茹[1,2] 王佩[1,2] 金鑫[1,2] 杨银凤[1,2] 

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]农业部动物疾病临床诊疗技术重点实验室,内蒙古呼和浩特010018

出　　处：《中国兽医学报》2016年第4期604-612,共9页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31160491)

摘　　要：为了解不同乳酸杆菌的刺激对绵羊瘤胃上皮细胞中绵羊β-防御素-1(sheep beta-defensin-1,SBD-1)的可能调节作用,本研究选用了2株具有益生作用的乳酸杆菌,即干酪乳杆菌Zhang(L.casei Zhang)和植物乳杆菌P-8(L.plantarum P-8),首先,运用实时荧光定量PCR技术(RT-q PCR)检测与乳酸杆菌共培养后的绵羊瘤胃上皮细胞中SBD-1mRNA的相对表达量;其次,探讨了2株乳酸杆菌诱导SBD-1 mRNA表达的差异性,选取优势菌株;最后,运用RTq PCR技术和酶联免疫吸附测定(ELISA)的方法,检测优势菌株活菌、灭活菌及其培养上清液对SBD-1基因及蛋白水平表达的影响。结果表明:2株乳酸杆菌均能促进SBD-1 mRNA的相对表达量,乳酸杆菌的浓度为107CFU/m L、刺激细胞8 h时能极显著促进SBD-1 mRNA的相对表达量(P<0.01),而L.plantarum P-8强于L.casei Zhang;进一步的研究表明L.plantarum P-8的灭活菌亦可显著诱导SBD-1的基因及蛋白表达(P<0.05),但弱于活菌。本研究结果表明L.plantarum P-8和L.casei Zhang均可诱导绵羊瘤胃上皮细胞内SBD-1的差异性表达。In order to explore the possible effect of different lactobacilli on the β-defensin -1 （sheep beta-defensin-1, SBD-1） expression in cultured ruminal epithelial cells of sheep. Firstly, two strains of probiotic lactobacilli ,L. casei Zhang and L. plantarum P-8 were chosen to co-culture with rumi- nal epithelial cells of sheep,then the relative expression of SBD-1 mRNA was determined by RT- qPCR experiments. Secondly,the dominant strain was selected by the effect of two lactobacilli on the SBD-1 expression. Finally,epithelial cells were stimulated with active,inactive and culture su- pernatant of the selected lactobacillus, the expression of SBD-1 mRNA and protein were deter- mined by RT-qPCR and ELISA, respectively. The results showed that the relative expression of SBD-1 mRNA was induced by two strains of lactobacilli and had a very significant difference when the concentration of the lactobacilll was 107 CFU/mL for 8 h （P〈0.01）, but L. plantarum P-8 was stronger than L. casei Zhang. Further studies showed that inactived L. plantarum P-8 can also significantly induced SBD-1 gene and protein expression （P〈0. 05） ,but weaker than live. The re- sults of this study indicated that L. plantarum P-8 and L. casei Zhang can induce different expres- sion of SBD-1 in ruminal epithelial cells of sheep.

关 键 词：β-防御素-1 瘤胃上皮细胞 乳酸杆菌 实时荧光定量PCR 酶联免疫吸附测定 

分 类 号：S852.61[农业科学—基础兽医学]