高黏附性乳酸菌的筛选及其对免疫细胞活性影响的研究  

Screening of lactic acid bacteria with high adhesion and its effect on the activity of immune cells

作　　者：刘东方 薛梅 桑建 陈大卫 何勇 李正华 王春雷 顾瑞霞 

机构地区：[1]扬州大学江苏省乳品生物技术与安全控制重点实验室,江苏扬州225001 [2]统一企业(中国)投资有限公司,江苏昆山215300

出　　处：《中国微生态学杂志》2016年第8期869-874,共6页Chinese Journal of Microecology

基　　金：国家自然科学基金(31571855);"十二五"国家科技支撑计划项目(2013BAD18B12);江苏省科技支撑项目(BN2014080);江苏省高校自然科学研究重大项目(12KJA550003)

摘　　要：目的以实验室保藏的具有良好加工特性的20株乳酸菌为研究对象,研究它们表面疏水性、对大鼠小肠上皮细胞(IEC-6)黏附性以及对脾细胞和巨噬细胞免疫效力的影响。方法采用微生物粘着碳氢化合物法(bacteria adhesion to hydrocarbons,BATH)测定乳酸菌细胞表面疏水性;采用革兰染色法评价乳酸菌的黏附性能;采用4-甲基偶氮唑盐(MTT)染色法检测脾细胞增殖和巨噬细胞吞噬中性红能力;采用试剂盒测定脾细胞分泌肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)和巨噬细胞释放一氧化氮(NO)含量。结果菌株LV108、N4和XJH301的表面疏水性高于其他菌株(F=221.9680,P=0.0000),分别为54%、47%和44%,同时这3株菌对细胞的黏附菌数也高,均大于14个/细胞;菌株LV108可以显著诱导脾细胞增殖(F=125.0430,P=0.0000)和提高TNF-α和IL-2的释放量;菌株LV108可以显著增强巨噬细胞吞噬活性和提高NO分泌水平(F=101.0030,P=0.0000)。结论菌株LV108具有高黏附性能,可以激活免疫细胞及调节细胞因子分泌发挥一定的免疫增强作用。Objective To study the cell surface hydrophobicity of 20 strains of laboratory preserved lactic acid bacteria with good processability,their adhesion to rat IEC-6cells and effects on the immune efficacy of spleen cells and macrophages.Methods The surface hydrophobicity of lactic acid bacteria cells were measured with the method of BATH（bacteria adhesion to hydrocarbons）.Splenocyte proliferation and the uptake of neutral red by macrophages were determined with MTT staining method.The adhesion performance of lactic acid bacteria was evaluated with Gram staining method.The contents of TNF-α,IL-2and NO were determined with appropriate test kits respectively.Results The surface hydrophobicities of strains LV108,N4 and XJH301were higher than those of the other strains（F=221.9680,P=0.0000）,being 54%,47%and 44% respectively.The cell adhesion indexes of these three strains were also higher,over 14/cell.The strain LV108 significantly increased the proliferation of splenocytes（F=125.0430,P=0.0000）and the levels of TNF-αand IL-2,and it significantly improved the phagocytosis of neutral red by macrophages and the secretion of NO（F=101.0030,P=0.0000）.Conclusion Strain LV108 has high adhesion performance,which can play a role in immune enhancement by activating immune cells and regulating cytokines.

关 键 词：乳酸菌 免疫细胞 黏附 增殖 细胞因子 

分 类 号：R378.992[医药卫生—病原生物学]