乳杆菌对肠上皮样细胞HT-29的黏附特性

The adhesive characters of Lactobacillus to enterocyte-like HT-29 cells

目的探讨乳杆菌对肠上皮样细胞HT-29的黏附特性。方法采用细胞培养革兰染色法,观察pH、共孵育时间、菌液浓度、生长状态、D-甘露糖以及宿主细胞类型等因素对乳杆菌黏附HT-29细胞的影响。结果酸性环境有利于乳杆菌对HT-29细胞的黏附,乳杆菌在pH=4.0时的黏附数显著高于pH=6.0时。处于稳定生长期的乳杆菌的黏附性能显著优于对数生长期的乳杆菌。D-甘露糖可明显抑制乳杆菌对HT-29细胞的黏附。乳杆菌对细胞的黏附具有时间和浓度依赖性,且具有宿主细胞特异性。结论乳杆菌的黏附具有种属和宿主特异性,且受多种外部因素的影响。

Objective To study the adhesive characters of Lactobacillus to enterocyte-like HT29 cells. Methods A HT-29 cell culture was employed to investigate the adherent ability of Lactobacillus Bacterial suspension and HT-29 cells were co-incubated at 37℃ for lh. The cells with adherent bacteria were Gram-stained, and then examined microscopically under oil immersion. The pH of the medium, bacterial growth phase, co-incubation time, bacterial concentration, D-mannose and type of host cells, which were involved in the process of adherence of Lactobacillus to enterocyte-like HT29 cells, were also investigated. Results The adherent level increased substantially for LactobaciUus reuteri JCM1081 as the pH decreased from 6. 0 to 4. 0. Maximum adherence occurred within 90rain and remained stable until the end of incubation. The bacterial concentration exerted a certain influence on the adherence of Lactobacillus reuteri JCM1081 to HT29 cells, and it seemed to be concentration dependent. Maximum adherence occurred in the concentration of 10^9cfu/ml and remained stable until the end of incubation. Lactobacillus strain that grew into stationary phase showed higher ability to adhere to HT29 cells than that in logarithmic phase. The addition of D-mannose dramatically decreased the level of Lactobacillus adherence. The Lactbacillus strain adhered in higher levels to the enterocyte-like cells or gastric epithelium cells than other cell origins, and the adhesion of Lactobacillus showed host cells specificity. Conclusion The adhesive characters observed with Lactobacillus suggested that adhesive properties are species and host specific. The pH, bacterial growth phase, co-incubation time, bacterial concentration and D-mannose may influence the adhesion of Lactbacillus to enterocyte-like HT-29 cells.