植物乳杆菌NDC75017对Caco-2细胞中il-6表达的影响  

Lactobacillus plantarum NDC 75017 affects il-6 gene expression in Caco-2 cells

作　　者：刘颖[1] 满朝新[2] 吕学娜[1] 韩琳琳[1] 薛玉清[1] 杨相宜[1] 单艺[2] 张英华[1] 姜毓君[1,2] 

机构地区：[1]东北农业大学食品学院,乳品科学教育部重点实验室,哈尔滨150030 [2]东北农业大学,国家乳业工程技术研究中心,哈尔滨150086

出　　处：《微生物学报》2012年第10期1237-1243,共7页Acta Microbiologica Sinica

基　　金：国家自然科学基金(31171718);黑龙江省教育厅科学技术研究重点项目(12511z005);国家"863计划"(2011AA100902);长江学者和创新团队发展计划(IRT-0959-203);黑龙江省青年科学基金项目(QC2009C55)~~

摘　　要：【目的】本研究通过体外实验,分析植物乳杆菌NDC75017对免疫相关基因白细胞介素6(il-6)表达的影响,并进一步揭示其机制。【方法】植物乳杆菌NDC 75017作用于Caco-2细胞0、2、4、6、8、10和12 h,采用Real Time PCR方法检测il-6基因和toll样受体2(tlr2)的表达。植物乳杆菌NDC 75017与Caco-2细胞共培养0、0.5、1、2和4 h,用western blot方法检测NF-κB的磷酸化水平;NF-κB的特异性抑制剂PDTC预处理Caco-2细胞30 min后,再加入植物乳杆菌NDC 75017作用2 h,用Real Time PCR方法检测il-6基因及tlr2的表达量。【结果】植物乳杆菌NDC 75017诱导Caco-2细胞中il-6和tlr2基因的表达,并且分别在诱导8 h和6 h时表达量达到最大。植物乳杆菌NDC 75017能够快速诱导NF-κB的磷酸化作用,在加入其特异性抑制剂PDTC后,il-6和tlr2基因的表达显著下降。【结论】植物乳杆菌NDC 75017能通过tlr2介导的NF-κB信号通路来诱导细胞因子il6短暂性的表达。[Objective] We tested the expression of immune-related gene interleukin 6（il-6） in vitro to understand the influence from Lactobacillus plantarum NDC 75017 on host cells and further to reveal the regulatory mechanism.[Methods] Caco-2 cells were cocultured with L.plantarum NDC 75017 for 0,2,4,6,8,10 and 12 h,the total RNA were extracted;then the expressions of il-6 and tlr2 genes were analyzed by Real Time RT-PCR.The phosphorylation level of NF-κB was analyzed by Western Blot after the Caco-2 cells stimulation with L.plantarum NDC 75017 at 0,0.5,1,2 and 4 h.Caco-2 cells were pretreated with pyrrolidine dithiocarbamate for 30 min before being treated with L.plantarum NDC 75017 for 2 h,then the total RNA was extracted and the expressions of il-6 and tlr2 genes were analyzed by Real Time RT-PCR.[Results] Lactobacillus plantarum NDC 75017 could induce the expressions of il-6 and tlr2 in Caco-2 cells,the il-6 and tlr2 expressions peaked at 8 h and 6 h after cocultured with L.plantarum NDC 75017.L.plantarum NDC 75017 could rapidly activate the phosphorylation of NF-κB,and the expressions of il-6 and tlr2 were decreased notably after pretreated with pyrrolidine dithiocarbamate.[Conclusion] L.plantarum NDC 75017 could up-regulate and then down-regulate the expression of il6 through rapidly activating tlr2-mediated NF-κB signaling pathway in Caco-2 cells.

关 键 词：植物乳杆菌NDC 75017 CACO-2细胞 白细胞介素6(il-6) toll样受体2(tlr2) NF-ΚB信号通路 

分 类 号：Q935[生物学—微生物学]