阴道乳杆菌菌种构成及其功能改变在VVC发生发展中的作用研究

Vulvovaginal candidiasis (VVC) is one of the most common causes of vaginitis,which seriously affected the health of many women.Previous studies have indicated that lactobacilli are the predominant microorganisms in vaginas of VVC. Lactobacilli consist several species, and the biological funcitons of each lactobacillus species are not the same.Our previous study showed that the different composition of lactobacillus species in vagina of women may determine its ability of regulating the balance of vaginal microbiota, and the composition type were closely related to the health of women.The present study would investigate the difference of composition of vaginal lactobacillus species between VVC and healthy women by using pyrosequencing, in order to assess the contribution of the composition of vaginal lactobacillus species to the onset of VVC.At the same time, bacteria culture would be used to assess the difference of biological function (production of lactic acid,ability of adhereing to the vaginal squamous epithelium,ability of inhibiting the growth of Candida albicans)of lactobacillus species between VVC and healthy women, to evaluate if the biological function of vaginal lactobacillus species were related to the onset of VVC.The present study would elucidate the relationship between the composition of vaginal lactobacillus species, and the relationship between the biological function of vaginal lactobacillus species and the onset of VVC. Meanwhile, the present study would supply with useful information for the diagnosis and treatments of VVC from the perspective of microecology.

外阴阴道假丝酵母菌病（VVC）是严重影响女性健康的常见病。研究表明：VVC患者阴道微环境中的优势菌亦为乳杆菌属。乳杆菌属有多个菌种，各菌种的生物功能存在差别。我们的前期研究发现，阴道微环境中乳杆菌菌种的不同组合（菌种构成）可能决定其调控阴道微生态平衡的能力，菌种构成与其维护生殖系统健康的能力存在密切相关性。本课题将采用以高通量测序技术为基础的宏基因组学方法，研究VVC 患者阴道微环境中乳杆菌菌种构成与健康女性阴道微环境中乳杆菌菌种构成的差别，分析乳杆菌菌种构成与VVC 发病之间的关系；同时采用体外培养法，研究阴道微环境中乳杆菌菌种的主要生物功能（产乳酸效率、粘附功能、抑制白色念珠菌增殖等）的差异，分析阴道乳杆菌菌种功能改变与VVC 发生之间的关系。本研究将为阐明阴道乳杆菌菌种构成和生物功能改变与VVC 发生发展之间的关系提供重要信息，为复发性及重度VVC 的微生态学诊断和治疗奠定基础。

外阴阴道假丝酵母菌病（VVC）是严重影响女性健康的常见病。研究表明：VVC患者阴道微环境中的优势菌亦为乳杆菌属。乳杆菌属有多个菌种，各菌种的生物功能存在差别。我们的前期研究发现，阴道微环境中乳杆菌菌种的不同组合（菌种构成）可能决定其调控阴道微生态平衡的能力，菌种构成与其维护生殖系统健康的能力存在密切相关性。.本研究中对180份标本（其中健康女性标本40例，单纯性VVC患者标本45例，RVVC患者标本45例，重度VVC患者标本50例）进行了多角度研究，结果如下：.1.共收集了180例符合研究要求的女性阴道分泌物标本，其中健康女性40例，单纯性VVC患者45例，RVVC患者45例，重度VVC患者50例。共获得了180份标本中的总细菌基因组DNA，同时成功分离了乳杆菌100株、白假丝酵母菌28株，并进行了鉴定和冻存。 .2.通过对180份标本的细菌基因组进行高通量测序，结果发现对照组的女性阴道微环境中乳杆菌多样性明显多于各研究组，其中95%的健康女性阴道中乳杆菌菌种为2-4种（其中12.5%有两个并存，37.5%有三个并存，35%有四个并存，10%有五个并存，2.5%的有六个并存），仅2.5%患者阴道中为1个菌种。而在研究组中，乳杆菌菌种多样性不同程度减低，可见单纯性VVC、复发性VVC和重度VVC中的仅含有一个菌种的比例分别为27%、35%和40%，而菌种在2-4个的比例降低到72%、65%和60%。.3.通过检测乳杆菌产生乳酸的种类及含量、乳杆菌抑制白假丝酵母菌增殖能力比较，发现所有乳杆菌均可产生两种乳酸，VVC组产生的乳酸总量明显少于对照组。VVC患者阴道乳杆菌的数量较正常组未见明显减少，但是乳杆菌菌种构成存在差别。在VVC组患者阴道乳杆菌的乳酸总产量低于对照组。VVC组患者阴道乳杆菌抑制白假丝酵母菌的功能弱于对照组。上述结果提示乳酸产生效率减少和抑菌功能减退可能参与了VVC的发生发展。