嗜酸乳杆菌黏附派伊尔结及抑制病原菌侵袭性的研究

Study on adhesion of LactobaciUus acidophilus to mice Peyer＇s patches and inhibition of pathogen invasion

目的研究嗜酸乳杆菌（Lactobacillus acidophilus）FN001黏附小鼠派伊尔结的黏附介导物，以及L.acidophilus FN001抑制病原菌黏附及侵袭派伊尔结的效果。方法通过化学、酶预处理研究黏附因子性质，通过单糖抑制黏附的性质研究黏附特异性。利用3种竞争黏附试验研究L．acidophilusFN001抑制病原菌黏附的作用，利用小肠结扎段法及电镜观察L．acidophilusFN001抑制大肠杆菌侵袭派伊尔结的效果。结果蛋白酶处理可以显著降低黏附，高碘酸处理可以显著增强黏附，甘露糖和甲基-α—D-甘露糖可以显著抑制黏附，L.acidophilusFN001可不同程度抑制病原菌黏附派伊尔结，对大肠杆菌效果最佳。结论L．acidophilusFN001通过蛋白样物质利用甘露糖特异方式黏附大鼠派伊尔结。L．acidophilusFN001可抑制通过相似方式黏附派伊尔结的病原菌。

Objective To investigate the adhesion mechanism of LactobaciUus acidophilus FN001 to Peyer＇s patches. Methods Adhesion of L. acidophilus FN001 to mice Peyer＇s patches was studied in vitro using a fluorescent quantization method. The nature of adhesion mediator was studied by the effects of physical, chemical and enzymatic pre-treatments of the bacteria on their adhesion and effect of sugars on inhibition of adhesion. The presence of lectin-like proteins in the cell surface was determined by hemagglutination. Effect of L. acidophilus FN001 on inhibition of adhesion of pathogens to Peyer＇s patches was also studied. Results The adhesion of L. acidophilus FN001 was strongly inhibited in the presence of D-mannose and methyl-α-D-mannoside. Pretreatment of L. acidophilus FN001 with pepsin and trypsin decreased the ad- hesive capacity indicating that cell surface proteins are involved in adhesion to Peyer＇s patches. L. acidophilus FN001 could agglutinate rabbit red cell in mannose specific manner and protease pretreatment could decrease hemagglutinin, suggesting that L. acidophilus FN001 has mannose specific lectin （s）. In adherence inhibition assay, L. acidophilus NF001 could significantly inhibit adhesion of E. coli ATCC25922 to Peyer＇s patches when L. acidophilus NF001 were applied to Peyer＇s patches first or at the same time with pathogen. Conclusion It was concluded that a mannose-specific protein mediated adhesion of L. acidophilus FN001 to the Peyer＇s patches, and L. acidophilus FN001 could inhibit adhesion of pathogen with similar lectins specificity to Peyer＇s patches.