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儿茶素对肠出血性大肠杆菌的杀菌及抗毒素活性

[Bactericidal and anti-toxin activities of catechin on enterohemorrhagic Escherichia coli].

作者信息

Okubo S, Sasaki T, Hara Y, Mori F, Shimamura T

机构信息

Department of Microbiology and Immunology, Showa University School of Medicine.

出版信息

Kansenshogaku Zasshi. 1998 Mar;72(3):211-7. doi: 10.11150/kansenshogakuzasshi1970.72.211.

Abstract

We examined the bactericidal activity of catechin, an astringent ingredient of tea, on enterohemorrhagic Escherichia coli (EHEC) O157:H7 and the anti-toxin activity of catechin on vero toxin (VT), the main pathogenic factor of EHEC O157:H7. To examine bactericidal activity, we added 1 X 10(4) CFU/ml bacteria to 1.25 to 20 W/V% of green tea extract or the PBS solution containing 25 to 400 micrograms/ml of (-) epigallocatechin gallate (EGCg), which is the main catechin ingredient of green tea leaf, and counted the number of live bacteria at various intervals. After 3 to 5 hours, no live bacteria were seen in 1.25 to 2.5 (regular drinking concentration) % green tea extract. In the high concentrations of 100 to 400 micrograms/ml EGCg the number of live bacteria decreased with time and after 24 hours no survivors were seen. In the low concentrations of 25 to 50 micrograms/ml EGCg, however, no change was observed in the number of live bacteria during 5 hours. After 24 hours the bacteria in 50 micrograms/ml were killed and the number of bacteria in 25 micrograms/ml decreased to one tenth of that at the start. To examine the anti-toxin activity, we mixed equal volumes of 2 ng/0.1 ml VT2 and 0.5 to 2 mg/0.1 ml catechin in vitro and incubated them at 37 degrees C for various times. Then we inoculated 0.2 ml of the mixture intraperitonealy to BALB/c mice. One mg of catechin inhibited by 100% the lethal toxicity of 2 ng of VT2 (LD 100) to mice. The inhibition of lethal toxicity of VT2 by catechin depended on the incubation time. The rate of inhibition was 0, 40 and 100% for 9, 12 and 18-24 hours incubation, respectively. These results suggest that catechin has not only bactericidal activity on EHEC O157:H7 but also anti-toxin activity on vero toxin.

摘要

我们研究了茶中的收敛成分儿茶素对肠出血性大肠杆菌(EHEC)O157:H7的杀菌活性,以及儿茶素对EHEC O157:H7的主要致病因子志贺毒素(VT)的抗毒素活性。为了检测杀菌活性,我们将1×10(4) CFU/ml的细菌加入到1.25至20 W/V%的绿茶提取物或含有25至400微克/毫升(-)表没食子儿茶素没食子酸酯(EGCg)的PBS溶液中,EGCg是绿茶叶片中的主要儿茶素成分,并在不同时间间隔计数活菌数量。3至5小时后,在1.25至2.5%(常规饮用浓度)的绿茶提取物中未见活菌。在100至400微克/毫升的高浓度EGCg中,活菌数量随时间减少,24小时后未见存活菌。然而,在25至50微克/毫升的低浓度EGCg中,5小时内活菌数量未见变化。24小时后,50微克/毫升中的细菌被杀死,25微克/毫升中的细菌数量降至开始时的十分之一。为了检测抗毒素活性,我们在体外将等体积的2纳克/0.1毫升VT2和0.5至2毫克/0.1毫升儿茶素混合,并在37℃下孵育不同时间。然后将0.2毫升混合物腹腔注射到BALB/c小鼠体内。1毫克儿茶素可100%抑制2纳克VT2对小鼠的致死毒性(LD 100)。儿茶素对VT2致死毒性的抑制作用取决于孵育时间。孵育9、12和18至24小时的抑制率分别为0、40%和100%。这些结果表明,儿茶素不仅对EHEC O157:H7具有杀菌活性,而且对志贺毒素具有抗毒素活性。

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