阴道乳杆菌与宫颈癌相关性研究进展

人类阴道微生物群是由200多种细菌组成的复杂系统,其组成受到基因、民族背景、行为以及环境因素等的影响。其中乳杆菌是女性阴道内的优势菌,在健康女性的阴道内主要有L. iners、L. crispatus、L. gasseri和L. jensenii这4种乳杆菌。阴道内乳杆菌主要通过代谢阴道上皮细胞内的糖原产生乳酸,维持阴道正常的酸性环境。此外,阴道乳杆菌还可通过产生过氧化氢(H2O2)和细菌素等物质发挥抵御致病微生物入侵的作用。近年来的研究显示女性阴道HPV感染与阴道微生态失衡尤其是乳杆菌含量的减低有关。本文将围绕阴道内乳杆菌的生理作用及其与宫颈癌发病的相关性作一综述。     

乳杆菌生态消毒剂对阴道毛滴虫的抑制作用研究

目的:在实验室对以乳杆菌发酵液为主要成分的乳杆菌生态消毒剂进行杀灭阴道毛滴虫效果观察.方法:采用定量抑菌试验方法进行.结果:以该剂原液、5倍稀释液和10倍稀释液对阴道毛滴虫作用4 h以上,对滴虫的抑制率都超过90%以上.结论:伊清爽乳杆菌阴道生态消毒剂具有抑制阴道毛滴虫的作用,是新一代保健型阴道卫生用品.     

妊娠期妇女阴道微生态研究进展

阴道微生态是人体系统中的一个非常敏感而又复杂的系统,受各种因素的影响。而在妊娠期妇女因其生理变化,会对阴道微生态产生一定影响。本研究总结迄今为止对于妊娠期妇女阴道微生态状况的分析和见解,阐明妊娠期妇女阴道环境的特点及病因,并对妊娠期妇女阴道微生态的相关疾病作一综述。     

乳杆菌对全子宫切除术后阴道微生态的影响

目的 分析甲硝唑阴道准备联合全子宫切除术对患者阴道微生态的影响及乳杆菌的应用价值。 方法 采用前瞻性分析,收集2015年1月至2018年5月我院符合本研究相关标准并行腹腔镜全子宫切除的患者100例为研究对象,患者术前均用甲硝唑进行阴道准备。采用随机数字表将入选患者分为2组,每组50例。观察组患者在术后2个月时采用乳杆菌活菌胶囊阴道给药,1次/d,2粒/次,10 d/疗程,共3个疗程;对照组不处理。观察2组患者术前、术后2个月、术后3个月时阴道乳杆菌数量、pH值、菌群多样性、菌群密集度分级差异,同时对比观察组与对照组患者治疗前后阴道乳杆菌检出率、阴道菌群失衡率、pH值、FSFI分值。 结果 (1)100例患者术后2个月时阴道乳杆菌数量、pH值、菌群多样性、菌群密度分级均显著减少,以Ⅰ级为主;术后3个月时以Ⅳ级为主。(2)与治疗前相比,两组患者阴道乳杆菌检出率均升高,阴道菌群失衡率均降低,pH值均降低,FSFI分值均升高(均P0.05);治疗后观察组患者阴道乳杆菌检出率高于对照组,阴道菌群失衡率、pH值低于对照组,FSFI分值高于对照组(均P2=3.78,P=0.022)。 结论 甲硝唑阴道准备联合腹腔镜全子宫切除术患者术后阴道微生态环境被破坏,尤其以术后2个月最为明显,给予乳杆菌干预能促使被破坏的阴道微生态环境恢复正常,减少术后细菌性阴道病的发生,值得应用和推广。     

乳杆菌制剂治疗外阴阴道念珠菌病的研究进展

外阴阴道念珠菌病(vulvovaginal candidiasis, VVC)又称念珠菌性阴道炎,是由念珠菌异常增殖引起的女性常见的第二大外阴阴道炎症疾病,常伴有阴道菌群失调和微环境的改变,尤其在妊娠期间,VVC的发病率升高,与不良妊娠结局的发生显著相关,对女性身心健康造成了严重的损害。传统针对VVC的治疗一直以抗真菌药物为主,但并未修复受损的阴道微生态系统,目前乳杆菌制剂被认为可通过维持正常的阴道微生物群来重建阴道微生态系统,有利于治疗女性阴道炎症疾病,因而在临床诊疗工作中,乳杆菌制剂的应用越来越多。以下就国内外乳杆菌制剂联合其他药物治疗VVC的研究进行综述,探讨乳杆菌制剂在VVC的治疗中发挥的作用。     

阴道常见乳杆菌及相关研究进展

近年来,随着全基因组测序等分子生物技术的应用,人们对阴道微生态系统的结构和功能的认识逐渐清晰。研究发现很多妇科疾病的出现伴随着阴道菌群失调,因此研究正常的阴道菌群结构和功能对预防和治疗菌群失调引起的疾病有重要意义,这也是目前人体微生态研究领域的热点。本文总结了近年关于健康女性阴道菌群的组成和类型的研究,阐述了阴道中常见乳杆菌如卷曲乳杆菌、惰性乳杆菌、格氏乳杆菌、詹氏乳杆菌等在功能研究方面的最新进展,以期为乳杆菌在防治妇科疾病如细菌性阴道病(BV)、外阴假丝酵母菌病(VVC)并推进其产品开发及临床应用等方面提供一些理论参考。     

阴道微生态亚失调状态的研究

目的 :提出阴道微生态亚失调状态的概念及其特征与机制。方法 :选取门诊及妇女健康普查患者 2 0 9例。健康妇女 10 2例为对照组 ;以临床主治外阴不适或偶有瘙痒 ,阴道分泌物增多 ,分泌物检查排除特异性微生物感染及细菌性阴道炎、严重宫颈炎的患者 10 7例为观察组 ;取阴道分泌物经倍比稀释接种于各培养基上进行需氧及厌氧培养 ,鉴定并检测微生物种类 ,活菌计数 ,检测菌种为乳酸杆菌、白色念珠菌、类杆菌、大肠埃希菌、链球菌 ,并同时测定阴道 p H值及清洁度。结果 :p H值无显著变化 (P>0 .0 5 ) ,清洁度观察组变化明显 ( °～ °) ;各菌种活菌计数观察组与对照组相比变化显著 (P>0 .0 5 )。结论 :阴道微生境在内外因素作用下存在一种过渡状态 ,该状态呈可逆性 ,在机体抵抗力增加或 /和去掉诱因后可自然恢复 ,如进一步发展可导致特异性或非特异性阴道炎、宫颈炎。作者由此提出“阴道微生态亚失调状态”概念。     

滴虫性阴道炎患者阴道乳杆菌功能的改变与重建

探讨滴虫性阴道炎(TV)患者阴道乳杆菌的功能状态及口服甲硝唑治疗后阴道乳杆菌功能的重建情况, 为TV的乳杆菌补充治疗提供依据。

选择TV患者及健康非孕女性各30例, 采用前后对照、标准对照的方法, 检测患者治疗前后阴道乳杆菌功能(定植密度、乳杆菌分级、产酸能力、产H₂O₂能力、体外增殖能力), 研究TV患者阴道乳杆菌功能的变化和重建情况。

TV患者阴道乳杆菌定植密度为(16.90±7.51)个/HP, 乳杆菌分级正常率为26.67%, H₂O₂阳性率为60.00%, 体外培养24 h pH值为4.71, 24 h菌液浊度为42.9 MCF。TV患者治愈后阴道乳杆菌定植密度为(32.09±8.79)个/HP, 乳杆菌分级正常率为39.13%, H₂O₂阳性率为60.87%, 体外培养24 h pH值为4.62, 24 h菌液浊度为45.6 MCF。TV患者治疗前阴道乳杆菌定植密度显著减少, 乳杆菌分级异常, 产H₂O₂能力、产酸能力、体外增殖能力显著减弱(均P < 0.05)。患者治愈后阴道乳杆菌定植密度显著增加(P < 0.05), 但未恢复正常, 乳杆菌分级、产H₂O₂能力增加不明显, 产酸能力、体外增殖能力较治疗前增强, 但均未恢复正常。

TV患者阴道乳杆菌功能减退, 治愈后未能完全重建, 需帮助恢复阴道乳杆菌功能。

     

600例健康体检妇女阴道微生态状况分析

目的研究深圳宝安地区健康妇女阴道微生态状况。方法对2011年9月至2012年2月在深圳宝安区人民医院体检中心进行健康体检的600例妇女阴道分泌物进行检测,检测指标包括清洁度、念珠菌、滴虫、阴道酸碱度、唾液酸苷酶、白细胞酯酶、过氧化氢（H2O2）浓度。结果研究对象中阴道微生态正常328例（54．67％）,微生态失调者272例（45．33％）;阴道微生态失调者中,微生态功能检测阳性率及致病菌的检出率均明显大于微生态正常者,差异有统计学意义（P〈0．05）。结论深圳宝安地区健康体检妇女部分患有阴道炎或存在阴道微生态失调或亚失调状态;阴道微生态联合检测法简便、快速、价廉实用,值得推广。     

4816例妇科门诊妇女阴道微生态状况的分析

目的研究当前厦门地区妇科门诊妇女阴道微生态的状况。方法对2010年8月至12月于厦门市妇幼保健院妇科门诊就诊的4 816例患者阴道分泌物进行检查,检查指标包括清洁度、念珠菌、滴虫、唾液酸苷酶、白细胞酯酶、过氧化氢(H2O2)浓度。结果研究对象中患阴道炎比例高(54.92%),阴道微生态失调率更高(60.86%);正常体检的妇女部分患有阴道炎或存在阴道微生态失调。结论当前厦门地区妇科门诊患者的阴道微生态状况形势是严峻的;阴道微生态6项快测指标简便、快速、实用,值得推广。     

不同生理阶段、避孕方式和感染状态对阴道乳酸杆菌影响的研究

目的调查不同生理阶段、避孕方式、感染因素对女性阴道乳酸杆菌数量的影响。方法收集南昌大学第二附院妇产科就诊的176例妇女的主要资料并采用qPCR方法定量分析阴道分泌物中乳酸杆菌数量,分泌物涂片和革兰染色后在显微镜下观察记录其感染状态。结果非绝经期组不同年龄段乳酸杆菌数量差异无统计学意义,绝经期组乳酸杆菌数量减少;不同避孕方式组的阴道乳酸杆菌数量差异无统计学意义;细菌性阴道病和假丝酵母菌感染组阴道乳酸杆菌数量较正常组差异均有统计学意义。结论生理阶段、细菌性阴道病和假丝酵母菌感染是影响女性阴道乳酸杆菌数量的主要因素。     

Characterization of vaginal lactobacilli in women after kidney transplantation

Limited number of publications described vaginal microflora after kidney transplantation. Our PubMed search revealed only 18 publications including words “vaginal bacteria &; kidney transplant” in the period of 1978–2011. The aim of this study was to characterize lactobacilli isolated from vaginal swabs of women after kidney transplantation, compared with healthy women. Eighteen renal transplant recipients (mean age 36.1) and 20 healthy women (mean age 36.0) were evaluated. Lactobacilli were cultured on MRS and Columbia blood agars. Biochemical identification with API 50 CHL (bioMerieux, Marcy L’Etoile, France) and multiplex PCR according to Song et al. was performed. Lactobacilli were tested for production of H₂O₂. Minimal inhibitory concentrations (MICs) of selected antimicrobial agents were determined with E-tests (bioMerieux, Marcy L’Etoile, France) and interpreted with CLSI and EUCAST criteria. No bacterial vaginosis was found among studied women. Two strains of group I were identified as Lactobacillus delbrueckii; 18 strains as Lactobacillus gasseri and 15 strains as Lactobacillus crispatus. Only 3 strains from group II were not identified by species-specific mPCR. Group IV was represented with 2 unidentified strains. Vaginal lactobacilli isolated from healthy women represented more homogenous group compared with heterogenous renal transplant recipients. Biochemical identification of lactobacilli by API 50 CHL kits was concordant with mPCR results only in 7 cases (17.5%), all 7 strains were identified as L. crispatus. Majority (93%) of lactobacilli were H₂O₂ producers. All isolated lactobacilli (100%) demonstrated high resistance to metronidazole (MIC > 256 μg/ml). Only 2 strains resistant to vancomycin (MICs: 32 and 256 μg/ml respectively), in the study and control group, and one to moxifloxacin (MIC = 32 μg/ml), were found. Resistance to metronidazole and vancomycin was concordant in CLSI and EUCAST (2010) criteria. Although significant differences between lactobacilli isolated from vaginas of kidney transplant and healthy women were not demonstrated, we demonstrated strains resistant to metronidazole, vancomycin and moxifloxacin in groups of examined women. Our study was performed on a small group of kidney transplant recipients and further more detailed molecular studies on a larger group of patients are required to confirm our results.     

Characterization of vaginal lactobacilli coaggregation ability with Escherichia coli

The coaggregation abilities of probiotic strains might enable it to form a barrier that prevents colonization by pathogenic bacteria. In the present study, the characterization of the coaggregation ability of 19 vaginal lactobacilli was studied. Coaggregation ability of all lactobacilli with Escherichia coli ATCC 11229 was positive. Only the highest coaggregation percentage of Lactobacillus acidophilus S1 was obtained with E. coli ATCC 11229 under both aerobic (71%) and anaerobic conditions (62%). The coaggregation abilities of strains occurred higher at acidic pH than at basic pH values. Moreover, the coaggregation abilities of tested strains against E. coli decreased after heat treatment (70 or 85 °C). Also, the relationship between hydrophobicity and coaggregation of strains was found to be significant. The effect of sonication, some enzymes (lipase and pepsin) and sodium periodate on coaggregation ability of L. acidophilus S1, which is one of the highest potentials on coaggregation ability, was investigated. Sodium periodate did not have a significant effect on coaggregation ability of L. acidophilus S1. The sonicated cell showed lower coaggregation than the control, the supernatant fluid of this sonicated cells showed similar coaggregation ability to the control. Coaggregation abilities of bacteriotherapeutic lactobacilli with pathogenic bacteria can be used for preliminary screening in order to identify potentially probiotic bacteria suitable for human use against urogenital tract infections.     

Molecular identification of vaginal lactobacilli isolated from Bulgarian women

Lactobacilli play an important role in maintaining the vaginal health of women. The development of suitable bacterial replacement therapies for the treatment of vaginosis requires knowledge of the vaginal lactobacilli species representation. The aim of this study was to identify at the species level vaginal Lactobacillus isolates obtained from Bulgarian women in childbearing age by using different molecular methods. Twenty-two strains of lactobacilli isolated from vaginal samples were identified and grouped according to their genetic relatedness. A combined approach, which included amplified ribosomal DNA restriction analysis (ARDRA), ribotyping and polymerase chain reaction (PCR) with species-specific oligonucleotide primers was applied. All vaginal isolates were grouped into 5 clusters in␣comparison with a set of 21 reference strains based␣on the initial ARDRA results, which was then confirmed by ribotyping. Finally, the strains were subjected to PCR analysis with eight different species-specific primer pairs, which allowed most of␣them to be classified as belonging to one of␣the␣following species: Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus helveticus and Lactobacillus plantarum. In conclusion, this study suggests that the most straightforward identification strategy for vaginal lactobacilli would be grouping by ARDRA or ribotyping, followed by PCR specific primers identification at species level.     

Beneficial lactobacilli: effects on the vaginal tract in a murine experimental model

Vaginal probiotics containing lactic acid bacteria with activity towards pathogenic microorganisms that cause urogenital tract infections have been proposed as a valid strategy for their prophylaxis and therapy. A murine experimental model was set up to evaluate the colonization capability of beneficial human lactobacilli and their effects on the mouse vaginal mucosa and innate immune cells. Five Lactobacillus strains were intravaginally inoculated into previously estrogenized BALB/c mice. The significance of the effects observed in the vaginal tract was determined by analysis of variance using the general linear model. The numbers of viable vaginal lactobacilli were significantly higher at proestrous–estrous than those at the metaestrous–diestrous phase and decreased markedly on the days after inoculation. Lactobacilli inoculation did not cause cytological or histological modifications of the murine vaginal tract. Moreover, the intravaginal administration of Lactobacillus salivarius CRL (Centro de Referencia para Lactobacilos culture collection) 1328 and Lactobacillus gasseri CRL 1263 did not affect the amounts of granulocytes and macrophages present in vaginal washings. In conclusion, the results demonstrate that vaginal lactobacilli did not produce adverse effects on the murine vaginal tract. Therefore, they could be proposed as safe probiotic candidates to promote a balanced microbiota in the urogenital tract.     

Role played by lactobacilli in controlling the population of vaginal pathogens

The role of Lactobacillus species in the female urogenital tract as a barrier to infection is of considerable interest. These organisms are believed to contribute to the control of vaginal microbiota by competing with other microorganisms for adherence to epithelial cells and by producing antimicrobial compounds. These bactericidal compounds include organic acid, which lowers the vaginal pH, hydrogen peroxide, bacteriocin-like substances and possibly biosurfactants.     

Role of the biological properties of vaginal lactobacilli in colonization process

The biological properties of vaginal lactobacilli strains with different colonization capacity were studied. As revealed in this study, colonization depended on the characteristics of species, individual elements of intrageneric antagonism, adhesion, resistance to antibacterial factors and the capacity for producing lysozyme. These properties of lactobacilli ensured the possibility of their adaptation to the conditions of the environment, manifested by its bacterial contamination. The data obtained in this study may be used for the study of symbiotic relationships between nonpathogenic bacteria with the host organism.     

Influence of probiotic vaginal lactobacilli on in vitro adhesion of urogenital pathogens to vaginal epithelial cells

AIMS: Lactobacilli, the predominant micro-organisms of the vaginal microbiota, play a major role in the maintenance of a healthy urogenital tract by preventing the colonization of pathogenic bacteria. The aim of the present study was to assess the ability of four vaginal Lactobacillus strains, previously selected for their probiotic features, to block in vitro the adherence of three human urogenital pathogens to vaginal epithelial cells (VEC). METHODS AND RESULTS: Three types of assays were performed in order to determine the inhibitory effect of lactobacilli on adhesion of urogenital pathogens to VEC: blockage by exclusion (lactobacilli and VEC followed by pathogens), competition (lactobacilli, VEC and pathogens together) and displacement (pathogens and VEC followed by the addition of lactobacilli). Bacterial adhesion to VEC was quantified by microscopy (x1000) after Gram's stain. All the strains were able to inhibit by exclusion and competition the adhesion of Staphylococcus aureus to VEC but none was able to decrease the attachment of Escherichia coli by neither of the mechanisms assayed. Only Lactobacillus acidophillus CRL 1259 and Lactobacillus paracasei CRL 1289 inhibited the attachment of Group B streptococci (GBS) to VEC by exclusion and competition respectively. CONCLUSIONS: Lactobacillus of vaginal origin were able to inhibit the attachment of genitouropathogenic Staph. aureus and GBS to the vaginal epithelium. SIGNIFICANCE AND IMPACT OF THE STUDY: The results support the probiotic potential of these Lactobacillus strains as anti-infective agents in the vagina and encourage further studies about their capacity to prevent and manage urogenital tract infections in females.     

Analysis of vaginal lactobacilli from healthy and infected Brazilian women

Culture-dependent PCR-amplified rRNA gene restriction analysis and culture-independent (PCR-denaturing gradient gel electrophoresis) methodologies were used to examine vaginal lactobacilli from Brazilian women who were healthy or had been diagnosed with vulvovaginal candidiasis (VVC) or bacterial vaginosis. Only Lactobacillus crispatus was detected accordingly by both methods, and H(2)O(2)-producing lactobacilli were not associated with protection against VVC.