嗜酸乳杆菌活菌体和热灭活菌体对 IPEC-J2细胞的黏附和黏附拮抗效应

摘要

The adhesion and adhesion antagonistic effect of live and heat-killed Lactobacillus acidophilus on IPEC-J2 cells were studied in vitro. IPEC-J2 cells were treated with live and heat-killed Lactobacillus acidophi-lus,and then stained the cells and observed the adhesive property. Moreover,the inhibition ratio of live and heat-killed Lactobacillus acidophilus to inhibit the adhesion of Escherichia coli on IPEC-J2 cells through three treatments(competitive inhibition,exclusion inhibition and displacement inhibition)was assessed by eosin-methylene blue agar(EMB)culture medium. The results showed as follows:live and heat-killed Lactobacillus acidophilus all could adhere well to the IPEC-J2 cells. The number of heat-killed bacteria adhering to the per cell[(40±2)CFU]was higher than the number of live bacteria adhering to the per cell[(20±2)CFU]. The two forms of Lactobacillus acidophilus had a good ability to inhibit the Escherichia coli adhere to IPEC-J2 cells. In competitive and displacement inhibition experiments,live bacteria showed a significantly better inhibi-tion ratio than heat-killed bacteria(P<0.05),whereas,in exclusion inhibition experiment,heat-killed bacteria showed a significantly better inhibition ratio than live bacteria( P<0.05). In conclusion,live and heat-killed Lactobacillus acidophilus can adhere well to the IPEC-J2 cells,and are able to reduce the adhesion and inva-sion to IPEC-J2 cells by Escherichia coli. Heat-killed Lactobacillus acidophilus inhibits Escherichia coli adhe-sion mainly through competition for host cell-binding receptors. Apart from competition for host cell-binding re-ceptors,some antibacterial substances from live Lactobacillus acidophilus are able to inhibit the growth and re-production of Escherichia coli.%本试验旨在研究嗜酸乳杆菌活菌体和热灭活菌体在体外模拟条件下对 IPEC-J2细胞的黏附和黏附拮抗效应。用嗜酸乳杆菌活菌体和热灭活菌体处理 IPEC-J2细胞，染色后观察细菌对细胞的黏附性。另外，采用嗜酸乳杆菌活菌体和热灭活菌体，通过竞争抑制、排阻抑制和替换抑制3种不同的处理方式抑制大肠杆菌在 IPEC-J2细胞上的黏附，利用伊红美蓝琼脂培养基，检测嗜酸乳杆菌对大肠杆菌黏附的抑制率。结果表明：嗜酸乳杆菌活菌体和热灭活菌体都能很好地黏附到 IPEC-J2细胞上，每个细胞上黏附的热灭活菌数[（40±2）CFU]高于黏附的活菌数[（20±2）CFU]。2种状态的嗜酸乳杆菌都能很好地抑制大肠杆菌在 IPEC-J2细胞上的黏附。在竞争抑制和替换抑制试验中，活菌体的抑制率显著高于热灭活菌体（ P＜0．05）。然而，在排阻抑制试验中，热灭活菌体的抑制率显著高于活菌体（P＜0．05）。由此可知，嗜酸乳杆菌的活菌体和热灭活菌体都能很好地黏附到 IPEC-J2细胞上，抑制大肠杆菌对细胞的黏附和入侵，热灭活菌体可能主要通过竞争黏附位点来抑制大肠杆菌的黏附，而活菌体可能不只这一机制，还会在代谢过程中产生一些抑菌物质，抑制大肠杆菌的生长繁殖。