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目的:研究健康女性阴道乳杆菌对阴道常见致病菌的抑制作用及其机制。方法:选取北京大学第一医院体检中心18～45岁的育龄期健康女性17例,从其阴道分泌物中分离、纯化乳杆菌并进行鉴定。采用spot on lawn和双层牛津杯法筛选出对粪肠球菌、微小消化链球菌、大肠埃希菌和白假丝酵母菌有抑制作用的阴道乳杆菌菌株。测定乳杆菌的生长曲线及代谢物内的乳酸含量。将乳杆菌与白假丝酵母菌共培养,观察乳杆菌菌体和菌液粗提物在抑制白假丝酵母菌的过程中各自所起的作用。结果:17例健康女性阴道分泌物中分离出23株乳杆菌,其中6株具有抑菌作用,分别是加氏乳杆菌、詹氏乳杆菌、阴道乳杆菌、卷曲乳杆菌、发酵乳杆菌和德氏乳杆菌,其中加氏乳杆菌增殖能力较强,詹氏乳杆菌最弱。6株乳杆菌代谢物内乳酸的含量不同,发酵乳杆菌最高为(2.72±0.035)mg/ml,卷曲乳杆菌和德氏乳杆菌较低,分别为(2.14±0.035)mg/ml和(2.13±0.025)mg/ml。加氏乳杆菌、詹氏乳杆菌、阴道乳杆菌的菌液、菌体及菌液粗提物均可以抑制白假丝酵母菌的生长(P≤0.05);发酵乳杆菌和德氏乳杆菌的菌液和菌体可以抑制白假丝酵母菌的生长(P≤0.05);卷曲乳杆菌对白假丝酵母菌则无抑制作用。结论:健康女性阴道乳杆菌的多个菌株均具有抑制大肠埃希菌、粪肠球菌、微小消化链球菌或白假丝酵母菌的功能。詹氏乳杆菌粗提物中细菌素类物质和过氧化氢的联合作用可以抑制白假丝酵母菌,加氏乳杆菌和阴道乳杆菌主要依靠过氧化氢起作用。乳杆菌产生的乳酸没有发挥抑菌作用。
Objective: To study the inhibitory effect and mechanism of Lactobacillus vaginalis on common vaginal pathogens in healthy women. Methods: Seventeen healthy women of childbearing age from 18 to 45 years old at the First Hospital of Peking University Health Check Center were selected. Lactobacillus was isolated and purified from vaginal secretions. Spot on lawn and double-layer Oxford cup were used to screen out Lactobacillus vaginalis strains which inhibit Enterococcus faecalis, Peinococcus aureus, Escherichia coli and Candida albicans. The growth curve of Lactobacillus and the content of lactate in the metabolites were determined. Lactobacillus and Candida albicans were co-cultured to observe the role of Lactobacillus bacteria and crude extract of bacteria in the inhibition of Candida albicans in their role. Results: Twenty-three Lactobacilli were isolated from 17 healthy women’s vaginal secretions, of which 6 had antibacterial activity, which were Lactobacillus gasseri, Lactobacillus jensenii, Lactobacillus jessiensis, Lactobacillus curvatus, Lactobacillus fermentum and German Lactobacillus, which increased ability to multiply Lactobacillus, Lactobacillus jane weakest. Lactobacillus fermentum had the highest lactobacillus (2.72 ± 0.035) mg / ml, lactobacillus curvatus and Lactobacillus delbrueckii were 2.14 ± 0.035 mg / ml and 2.13 ± 0.025) mg / ml. Lactobacillus gasseri, Lactobacillus gasseri, Lactobacillus gasseri, Lactobacillus gasseri, Lactobacillus gasseri, Lactobacillus gasseri, Lactobacillus jessiensis and Lactobacillus vaginalis could all inhibit the growth of Candida albicans (P≤0.05) Bacteria and bacteria can inhibit the growth of Candida albicans (P≤0.05); Lactobacillus curvatus did not inhibit Candida albicans. CONCLUSIONS: Multiple strains of Lactobacillus from healthy women are capable of inhibiting Escherichia coli, Enterococcus faecalis, Streptococcus diurnum, or Candida albicans. The combined action of the bacteriocins and hydrogen peroxide in the crude extract of Lactobacillus jinnii can inhibit the growth of Candida albicans, Lactobacillus gasseri and Lactobacillus vaginalis mainly by hydrogen peroxide. Lactobacillus produced lactic acid did not play an inhibitory effect.