发酵乳酸杆菌F6对鸡小肠上皮细胞β-防御素-9基因表达的影响  

Effect of probiotic Lactobacillus fermentum F6 on β-defensin-9 expression in epithelial cells of chicken small intestine

作　　者：洪智敏[1,2] 张和平[3] 贾永杰[1,2] 刘思国[2] 黎观红[1] 

机构地区：[1]江西农业大学动物科技学院,江西南昌330045 [2]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001 [3]内蒙古农业大学乳品生物技术与工程教育部重点实验室,内蒙古呼和浩特010018

出　　处：《中国兽医学报》2012年第8期1142-1147,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30800791);江西省青年科学家培养对象计划资助项目(赣科发计字[2010]209号)

摘　　要：采用实时荧光定量PCR(fluorescence quantitative PCR,FQ-PCR)检测益生性发酵乳酸杆菌F6刺激鸡小肠上皮细胞后抗菌肽β-防御素-9(AvBD9)基因表达变化,为从益生菌与上皮细胞抗菌肽表达关系的新角度解析益生菌发挥益生作用的新途径和机制提供一定的基础及依据。利用不同剂量(2×105,2×106,2×107 CFU)的发酵乳酸杆菌F6分别刺激原代培养的鸡小肠上皮细胞4h,提取刺激后的细胞总RNA,反转录为cDNA,FQ-PCR检测抗菌肽AvBD9基因表达变化。结果表明,未受刺激的正常对照组也检测到AvBD9mRNA的表达,发酵乳酸杆菌F6能上调AvBD9基因表达。刺激组中AvBD9mRNA的表达在不同剂量组之间存在差异。2×105 CFU/mL组AvBD9mRNA的表达量极显著高于未受细菌刺激的对照组和2×106 CFU/mL组(P<0.01),显著高于2×107 CFU/mL组(P<0.05)。2×106 CFU/mL组和2×107 CFU/mL组AvBD9mRNA的表达量显著高于未受细菌刺激的对照组(P<0.05),但2×106 CFU/mL组和2×107 CFU/mL组之间无显著差异(P>0.05)。发酵乳杆菌F6与鸡小肠上皮细胞相互作用过程中可提高抗菌肽AvBD9mRNA的表达,且存在剂量依赖性。Probiotic Lactobacillus fermentum F6（L. fermentum F6） was selected to investigate the effects of lactoba cillus stimulation on the 13-defensin-9（AvBD9） mRNA expression in cultured chicken small intestinal epithelial cells. The chicken small intestinal epithelial cells were stimulated for 4 h by different concentrations of L. fermentum F6 （2 × 105 , 2 ×106 , 2 × 107 CFU/mL）, and then the RNA was extracied from the stimulated cells and reversely transcrip- ted into cDNA. The AvBD9 mRNA expression was determinted by fluorescence quantitative PCR（FQPCR）. The results showed that the AvBD9 mRNA expression was observed in the control ceils without bacterial stimili, but the AvBD9 mRNA expression was upregulated by stimulation of L. ferrnentum F6 at different concentrations and ex- pression difference was observed between treatments at three bacterial concentrations. The AvBD9 mRNA expression in ceils stimulated with 2 ×05 CFU/mL L. fermentum F6 was very significantly higher than that stimulated with 2 × 106 CFU/mL L. fermentum F6（P〈0.01） ,and also was significantly higer than that treated with 2 × 107 CFU/mL L. fermentum F6（P〈0.05）. The AvBD9 mRNA expressions in cells stimulated with 2 X 106 CFU/mL and 2 × 107 CFU/mL L. fermentum F6 were both significantly higher than that in control cells（P〈0.05）, but no significant difference was observed between 2 × 10^6 CFU/mL and 2 × 10^7 CFU/mL L. fermentum F6 treatments（P〉0.05）. In conclusion,the AvBD9 mRNA expression was upregulated by L. fermentum F6 stimulation and in dose-dependent manner.

关 键 词：β-防御素-9 鸡小肠上皮细胞 发酵乳酸杆菌F6 荧光定量PCR 

分 类 号：S831.1[农业科学—畜牧学]