Mohammedsaeed Walaa, McBain Andrew J, Cruickshank Sheena M, O'Neill Catherine A
Institute of Inflammation and Repair, the University of Manchester, Manchester, United Kingdom.
School of Pharmacy and Pharmaceutical Sciences, the University of Manchester, Manchester, United Kingdom.
Appl Environ Microbiol. 2014 Sep;80(18):5773-81. doi: 10.1128/AEM.00861-14. Epub 2014 Jul 11.
Few studies have evaluated the potential benefits of the topical application of probiotic bacteria or material derived from them. We have investigated whether a probiotic bacterium, Lactobacillus rhamnosus GG, can inhibit Staphylococcus aureus infection of human primary keratinocytes in culture. When primary human keratinocytes were exposed to S. aureus, only 25% of the keratinocytes remained viable following 24 h of incubation. However, in the presence of 10(8) CFU/ml of live L. rhamnosus GG, the viability of the infected keratinocytes increased to 57% (P = 0.01). L. rhamnosus GG lysates and spent culture fluid also provided significant protection to keratinocytes, with 65% (P = 0.006) and 57% (P = 0.01) of cells, respectively, being viable following 24 h of incubation. Keratinocyte survival was significantly enhanced regardless of whether the probiotic was applied in the viable form or as cell lysates 2 h before or simultaneously with (P = 0.005) or 12 h after (P = 0.01) S. aureus infection. However, spent culture fluid was protective only if added before or simultaneously with S. aureus. With respect to mechanism, both L. rhamnosus GG lysate and spent culture fluid apparently inhibited adherence of S. aureus to keratinocytes by competitive exclusion, but only viable bacteria or the lysate could displace S. aureus (P = 0.04 and 0.01, respectively). Furthermore, growth of S. aureus was inhibited by either live bacteria or lysate but not spent culture fluid. Together, these data suggest at least two separate activities involved in the protective effects of L. rhamnosus GG against S. aureus, growth inhibition and reduction of bacterial adhesion.
很少有研究评估局部应用益生菌或其衍生材料的潜在益处。我们研究了益生菌鼠李糖乳杆菌GG是否能在培养中抑制人原代角质形成细胞的金黄色葡萄球菌感染。当原代人角质形成细胞暴露于金黄色葡萄球菌时,孵育24小时后只有25%的角质形成细胞仍存活。然而,在存在10⁸CFU/ml活的鼠李糖乳杆菌GG的情况下,受感染角质形成细胞的存活率增加到57%(P = 0.01)。鼠李糖乳杆菌GG裂解物和培养上清液也对角质形成细胞提供了显著保护,孵育24小时后分别有65%(P = 0.006)和57%(P = 0.01)的细胞存活。无论益生菌是以活的形式、细胞裂解物形式在金黄色葡萄球菌感染前2小时或同时(P = 0.005)或感染后12小时(P = 0.01)应用,角质形成细胞的存活率都显著提高。然而,培养上清液只有在金黄色葡萄球菌感染前或同时添加才有保护作用。关于作用机制,鼠李糖乳杆菌GG裂解物和培养上清液显然都通过竞争排斥抑制金黄色葡萄球菌对角质形成细胞的黏附,但只有活菌或裂解物能取代金黄色葡萄球菌(分别为P = 0.04和0.01)。此外,金黄色葡萄球菌的生长受到活菌或裂解物的抑制,但不受培养上清液的抑制。总之,这些数据表明鼠李糖乳杆菌GG对金黄色葡萄球菌的保护作用至少涉及两种独立的活性,即生长抑制和细菌黏附减少。