论文部分内容阅读
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。各个菌株在不同的剂量下,具有不同作用。提示在应用益生菌治疗免疫等相关性疾病时,应该考虑不同菌株对免疫细胞的不同作用机制。
Objective To compare the immunomodulatory effects of five different probiotic strains with different dosages and to provide a basis for selecting appropriate strains for treatment. Methods Cord blood mononuclear cells (CBMC) isolated from umbilical cord blood of 7 healthy pregnant women were collected, respectively, with Bifidobacterium longum 6-1, Bifidobacterium infantis CGMCC313-1, Lactobacillus acidophilus YIT2004, Streptococcus faecalis YIT0072 and Clostridium butyricum CGMCC313-2 were cocultured for 24-36 h at a ratio of bacteria to CBMC of 2: 1 (low), 20: 1 (medium) and 200: 1 , While negative control (PBS) and positive control (lipopolysaccharide, LPS). The expression of CD4 and CD25 on the surface of CBMC in each group was detected by flow cytometry. The levels of IL-10, IL-12, IL-4, TGF-β1 and IFN-γ in the culture supernatant were detected by ELISA. Results (1) In addition to 200-200:1 Clostridium butyricum CGMCC313-2 strain can significantly improve the CBCD expression of CD4 CD25 (10.45 ± 3.16 vs 5.84 ± 2.32, P = 0.009) compared with the negative control group (PBS) The remaining probiotic strains had no significant difference in the expression of CD4 CD25 (P> 0.05). (2) Bifidobacterium longum 6-1 could stimulate CBMC to produce IL-10 and IFN-γ at medium and high doses, but had no obvious effect on producing IL-12. (3) Bifidobacterium infantis CGMCC313-1 strain could stimulate CBMC to produce IL-10 at all dosages, with no significant effect on IL-12 and IFN-γ production. (4) Clostridium butyricum CGMCC313-2 strain could stimulate CBMC to produce IL-10 at medium and high doses, with no significant effect on IL-12 and IFN-γ production. (5) Streptococcus faecalis YIT0072 could stimulate CBMC to produce IL-10, IL-12 and IFN-γ at low dose, but had no effect at high dose. (6) Lactobacillus acidophilus YIT2004 could stimulate CBMC to produce IL-10 at medium and high doses, and could stimulate CBMC to produce IFN-γ at medium dose without any effect on IL-12. (7) IL-4 and TGF-β1 were not detected in this study. CONCLUSIONS: Among the probiotic strains currently used in China, only C. butyricum CGMCC313-2 strain can significantly enhance the expression of CBMC in CD25.5.5 strains, all of which can stimulate CBMC to produce anti-inflammatory cytokines IL-10; Bifidobacterium longum 6-1 Streptococcus faecalis YIT0072 strain and Lactobacillus acidophilus YIT2004 strain can stimulate CBMC to produce Th1 cytokine INF-γ, and only Streptococcus faecalis YIT0072 strain can stimulate CBMC to produce IL-12. Each strain has different effects at different doses. Prompt application of probiotics in the treatment of immune and other related diseases, different strains should be considered different mechanisms of immune cells.