五种益生菌菌株对脐血单个核细胞免疫作用的实验研究  

The effects of 5 probiotic strains on cord blood monocular cells

作　　者：李志川[1] 郑跃杰[1] 马红玲[1] 王和平[1] 祖莹[1] 

机构地区：[1]深圳市儿童医院,广东深圳518026

出　　处：《中国微生态学杂志》2015年第6期642-646,共5页Chinese Journal of Microecology

基　　金：2010年深圳市科技计划项目(201003147)

摘　　要：目的比较不同剂量的5种不同益生菌菌株的免疫调节作用,为选择适当的菌株进行治疗提供依据。方法取7例健康孕妇的脐血分离出的脐血单个核细胞（cord blood monocular cells,CBMC）,分别与长双歧杆菌6-1株、婴儿双歧杆菌CGMCC313-1株、嗜酸乳杆菌YIT2004株、粪链球菌YIT0072株和酪酸梭状芽胞杆菌CGMCC313-2株,以菌和CBMC比例2∶1（低）、20∶1（中）和200∶1（高）共培养24~36 h,同时设阴性对照（PBS）和阳性对照（脂多糖,LPS）。然后采用流式细胞仪检测各组CBMC表面CD4、CD25分子表达情况,用ELISA方法检测培养上清中IL-10、IL-12、IL-4、TGF-β1和IFN-γ的水平。结果 （1）与阴性对照（PBS）组相比,除200∶1比例的酪酸梭状芽胞杆菌CGMCC313-2株能够显著提高CBMC表达CD4CD25（10.45±3.16 vs 5.84±2.32,P=0.009）以外,其余益生菌菌株对表达CD4CD25差异均无统计学意义（P〉0.05）。（2）长双歧杆菌6-1株在中高剂量下,能够刺激CBMC产生IL-10和IFN-γ,对产生IL-12无明显影响。（3）婴儿型双歧杆菌CGMCC313-1株在各个剂量下均能够刺激CBMC产生IL-10,对IL-12和IFN-γ产生无明显影响。（4）酪酸梭状芽胞杆菌CGMCC313-2株在中高剂量下,能够刺激CBMC产生IL-10,对IL-12和IFN-γ产生无明显影响。（5）粪链球菌YIT0072株在低中剂量下,能够刺激CBMC产生IL-10、IL-12和IFN-γ,而高剂量则无影响。（6）嗜酸乳杆菌YIT2004株在中高剂量下,能够刺激CBMC产生IL-10,在中剂量下,能够刺激CBMC产生IFN-γ,对IL-12无影响。（7）在本研究中均未能检测出IL-4和TGF-β1。结论在目前国内使用的益生菌菌株中,仅酪酸梭状芽胞杆菌CGMCC313-2株能够显著提高CBMC表达CD4CD25。5种菌株均能够刺激CBMC产生抗炎症因子IL-10;长双歧杆菌6-1株、粪链球菌YIT0072株和嗜酸乳杆菌YIT2004株能够刺激CBMC产生Th1型细胞因子INF-γ,仅粪链球菌YIT0072株能够刺激CBMC产生IL-12。各个菌株在�Objective To explore the immunomodulatory effects of five probiotic strains of different species. Methods Cord blood monocular cells（ CBMC） were isolated from seven healthy donors and co-cultured with5 different strains of probiotic bacteria（ Bifidobacterium longum 6-1,Bifidobacterium infant CGMCC313-1,Lactobacillus acidophilus YIT2004,Streptococcus faecalis YIT0072 and Clostridium butyricum CGMCC313-2） at different bacteria / CBMC ratios for 24- 36 hours. Incubation with phosphate buffer solution（ PBS） and lipopolysaccharide（ LPS） were used as the negative and positive control respectively. The expression of CD4＋CD25＋on CBMCs were determined by flow cytometry. The levels of IL-10,IL-12,IL-4,TGF-β1 and IFN-γ in culture supernatants were evaluated by enzyme linked immunosorbent assay（ ELISA）. Results（ 1） Clostridium butyricum CGMCC313-2 increased the expression of CD4＋CD25＋on CBMC（ 10. 45 ± 3. 16 vs 5. 84 ± 2. 32,P = 0. 009）,while the other 4probiotic strains had no such effects.（ 2） Bifidobacterium longum 6-1 at medium-high doses increased the levels of IL-10 and IFN-γ in culture supernatants while no effect on that of IL-12.（ 3） Bifidobacterium infant CGMCC313-1 at each dose increased the level of IL-10 while no effect on those of IL-12 and IFN-γ.（ 4） Clostridium butyricum CGMCC313-2 at medium-high doses increased the level of IL-10 while no effect on those of IL-12 and IFN-γ.（ 5）Streptococcus faecalis YIT0072 at low-midium doses increased the levels of IL-10,IL-12 and IFN-γ while the high dose didn＇t.（ 6） Lactobacillus acidophilus YIT2004 increased the levels of IL-10 at medium-high doses and that of IFN-γ at medium dose,but no effects on that of IL-12.（ 7） No IL-4 and TGF-β1 were detected in culture supernatants. Conclusion Different probiotic strains have different immunomodulatory effects on CBMC at different doses,which should be taken into consideration when used for treatment of immune-related diseases.

关 键 词：益生菌 脐血单个核细胞 

分 类 号：R378[医药卫生—病原生物学]