Lactobacillus delbrueckii ssp. bulgaricus B-30892 can inhibit cytotoxic effects and adhesion of pathogenic Clostridium difficile to Caco-2 cells.
Lactobacillus delbrueckii ssp. bulgaricus B-30892 can inhibit cytotoxic effects and adhesion of pathogenic Clostridium difficile to Caco-2 cells.
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DOI：
10.1186/1757-4749-1-8
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发表时间：
2009-04-27
期刊：
影响因子：
4.2
通讯作者：
Bhunia AK
中科院分区：
文献类型：
--
作者：
Banerjee P;Merkel GJ;Bhunia AK
益生菌微生物在预防、治疗或饮食管理某些疾病（包括腹泻相关性腹泻（AAD））中的用途正受到越来越多的关注。艰难梭菌是AAD最常见的原因，并且所产生的C.艰难梭菌介导的感染（CDI）具有潜在的致命性。C.艰难梭菌相关性腹泻（CDAD）表现为严重的炎症和结肠炎，主要是由于C.引起肠上皮细胞破坏的艰难杆菌。本研究的目的是确定益生菌德氏乳杆菌ssp.保加利亚亚种B-30892（LD B B-30892）对C.使用Caco-2细胞作为模型的艰难介导的细胞毒性。本实验研究了C.来自LD B B-30892的不同稀释度（1：2至1：2048）的无细胞上清液（CFS）可改变艰难条件培养基对Caco-2细胞的作用。在类似的实验装置中，通过将来自这些菌株的结果与来自LD B B-30892的结果进行对比，特别是影响C的能力，对其他益生菌菌株进行比较评估。艰难梭菌对Caco-2单层诱导的细胞毒性。进行粘附测定，然后通过Giemsa染色进行定量分析，以测试来自LD B B-30892和其它益生菌测试菌株的CFS是否具有改变C的粘附的能力。Caco-2细胞单层的表达。还进行了实验以评价LD B B-30892或其释放组分是否对C有任何杀菌作用。很难LD B B-30892与C. difficile对C.艰难梭菌介导的对Caco-2细胞的细胞毒性。当CFS来自LD B B-30892-C时。在Caco-2单层上施用艰难梭菌共培养物（高达1：16的稀释度），没有细胞毒性的迹象。当来自单独培养的LD B B-30892的CFS与单独培养的C. LD B B-30892 CFS对C.以1：8的比例（LD B B-30892 CFS：C. difficile CFT）。我们没有发现任何类似的抑制C。当其他益生菌与LD B B-30892平行检测时，艰难梭菌介导的细胞毒性。我们的细胞毒性实验数据表明，LD B B-30892释放一种或多种生物活性成分到CFS中，从而中和由C诱导的细胞毒性。艰难梭菌，可能通过灭活其毒素。我们的数据还表明，来自LD B B-30892的CFS降低了C的粘附。difficile的益生菌增加了81%，这显著（P <0.01）高于本研究中测试的所有其他益生菌生物。这项研究揭示了德氏乳杆菌ssp.保加利亚乳杆菌B-30892（LD B B-30892）对C.艰难介导的细胞毒性，使用Caco-2细胞作为模型。研究还表明LD B B-30892可以减少C.结肠直肠细胞中的艰难梭状细胞。需要更多的研究来阐明这种细胞毒性和定殖减少的具体作用机制。
Probiotic microorganisms are receiving increasing interest for use in the prevention, treatment, or dietary management of certain diseases, including antibiotic-associated diarrhea (AAD). Clostridium difficile is the most common cause of AAD and the resulting C. difficile – mediated infection (CDI), is potentially deadly. C. difficile associated diarrhea (CDAD) is manifested by severe inflammation and colitis, mostly due to the release of two exotoxins by C. difficile causing destruction of epithelial cells in the intestine. The aim of this study was to determine the effect of probiotic bacteria Lactobacillus delbrueckii ssp. bulgaricus B-30892 (LDB B-30892) on C. difficile-mediated cytotoxicity using Caco-2 cells as a model. Experiments were carried out to test if the cytotoxicity induced by C. difficile-conditioned-medium on Caco-2 cells can be altered by cell-free supernatant (CFS) from LDB B-30892 in different dilutions (1:2 to 1:2048). In a similar experimental setup, comparative evaluations of other probiotic strains were made by contrasting the results from these strains with the results from LDB B-30892, specifically the ability to affect C. difficile induced cytotoxicity on Caco-2 monolayers. Adhesion assays followed by quantitative analysis by Giemsa staining were conducted to test if the CFSs from LDB B-30892 and other probiotic test strains have the capability to alter the adhesion of C. difficile to the Caco-2 monolayer. Experiments were also performed to evaluate if LDB B-30892 or its released components have any bactericidal effect on C. difficile. Co-culturing of LDB B-30892 with C. difficile inhibited the C. difficile-mediated cytotoxicity on Caco-2 cells. When CFS from LDB B-30892-C. difficile co-culture was administered (up to a dilution of 1:16) on Caco-2 monolayer, there were no signs of cytotoxicity. When CFS from separately grown LDB B-30892 was mixed with the cell-free toxin preparation (CFT) of separately cultured C. difficile, the LDB B-30892 CFS was inhibitory to C. difficile CFT-mediated cytotoxicity at a ratio of 1:8 (LDB B-30892 CFS:C. difficile CFT). We failed to find any similar inhibition of C. difficile-mediated cytotoxicity when other probiotic organisms were tested in parallel to LDB B-30892. Our data of cytotoxicity experiments suggest that LDB B-30892 releases one or more bioactive component(s) into the CFS, which neutralizes the cytotoxicity induced by C. difficile, probably by inactivating its toxin(s). Our data also indicate that CFS from LDB B-30892 reduced the adhesion of C. difficile by 81%, which is significantly (P <0.01) higher than all other probiotic organisms tested in this study. This study reveals the very first findings that Lactobacillus delbrueckii ssp. bulgaricus B-30892 (LDB B-30892) can eliminate C. difficile-mediated cytotoxicity, using Caco-2 cells as a model. The study also demonstrates that LDB B-30892 can reduce the colonization of C. difficile cells in colorectal cells. More study is warranted to elucidate the specific mechanism of action of such reduction of cytotoxicity and colonization.
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3.1
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