Isolation of adhesive strains and evaluation of the colonization and immune response by Lactobacillus plantarum L2 in the rat gastrointestinal tract.
Isolation of adhesive strains and evaluation of the colonization and immune response by Lactobacillus plantarum L2 in the rat gastrointestinal tract.
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DOI：
10.1016/j.ijfoodmicro.2009.03.016
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发表时间：
2009-06
影响因子：
5.4
通讯作者：
Bin Wang;Jie‐shou Li;Qiurong Li;Haiyun Zhang;Ning Li
中科院分区：
文献类型：
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作者：
Bin Wang;Jie‐shou Li;Qiurong Li;Haiyun Zhang;Ning Li
以Caco-2和IEC-6细胞为体外模型，检测了5株乳杆菌对人外周血单个核细胞(PBMC)分泌促炎和抗炎细胞因子的能力。在供试菌株中，植物乳杆菌L2对Caco-2和IEC-6细胞的粘附率最高，分别为595±125和704±273。此外，植物乳杆菌L2还能诱导PBMC产生相当水平的IL-10，但三种促炎细胞因子肿瘤坏死因子-α、干扰素-γ和IL-12的水平都很低。从这些结果中，选出了一株有希望的菌株，植物乳杆菌L2，用于体内研究。F344大鼠每天按2×109L剂量饲喂，连续28天。Plantarum L2；在接下来的14天里，大鼠不喂任何乳杆菌。分别于0、28、42d取肠黏膜标本和粪便，测定乳酸菌的定植能力。用API50CHL和菌株特异性聚合酶链式反应初步鉴定回收的乳酸菌。用菌株特异性分子探针进行荧光原位杂交分析肠道标本，通过免疫染色确定免疫细胞数量，以确定定植部位的免疫反应证据。大鼠摄入L_2 28天后，其粪便、小肠和结肠中的植物乳杆菌活菌数显著增加。即使在停止给药两周后，细菌水平仍然很高。菌株特异性的聚合酶链式反应和荧光原位杂交为植物乳杆菌L2在大鼠胃肠道的定植提供了明确和直接的证据。此外，在摄取植物乳杆菌L2后，回肠中CD19阳性细胞显著增加。综上所述，补充植物乳杆菌L2诱导了大鼠胃肠道的显著定植，这与胃肠道粘膜免疫反应的显著改变有关。
Five Lactobacillus strains were tested for their ability to adhere to Caco-2 and IEC-6 cell lines as in vitro models and to induce of the secretion of pro- and anti-inflammatory cytokines by human peripheral blood mononuclear cells (PBMCs). Among the tested strains, Lactobacillus plantarum L2 was the most adhesive strain, approximately 595±125 or 704±273 of the added bacteria adhered to Caco-2 or IEC-6 cell cultures, respectively. Furthermore, L. plantarum L2 was also found to induce a considerable level of IL-10 from PBMCs, but low levels of all three pro-inflammatory cytokines TNF-α, IFN-γ and IL-12. From these results, one promising strain, L. plantarum L2, was selected for in vivo studies. For 28 days F344 rats were fed a daily dose of 2×109L. plantarum L2; for the next 14 days the rats were not fed any Lactobacillus. Intestinal mucosal samples and feces were taken at days 0, 28 and 42 to determine the colonizing ability of the lactobacilli. Recovered Lactobacillus isolates were initially identified by API 50CHL and strain-specific PCR. Intestinal specimen was analyzed using fluorescence in situ hybridization with a strain-specific molecular probe, and immune cell populations were determined by immunostaining for evidence of immune responses at the colonized sites. After intake of L. plantarum L2 for 28 days, a significant increase in live L. plantarum was found in the rats' feces, small intestine and colon. The bacterial levels remained high even after the L. plantarum L2 administration had been stopped for two weeks. Strain-specific PCR and FISH provided clear and direct evidence of colonization of the rat gastrointestinal tract by L. plantarum L2. Additionally, a significant increase in CD19-positive cells in the ileum was observed after intake of L. plantarum L2. In conclusion, dietary supplementation with L. plantarum L2 induced significant colonization of the gastrointestinal tract of rats, and this was associated with significant alteration of the immune response in the gastrointestinal mucosa.