小鼠阴道菌群紊乱模型的建立

Establishment of mouse model of vaginal flora imbalance

目的建立小鼠阴道菌群紊乱模型,为调整阴道菌群紊乱的药物研发提供模型参考。方法采用雌激素化的C57BL/6小鼠经阴道以不同处理方式(链霉素50μg/只,加德纳菌50μL/只,链霉素50μg/只+抗链霉素加德纳菌50μL/只,每种处理方式各70只/组)诱导小鼠阴道菌群紊乱模型,采用细菌16S rDNA高通量测序考察造模后第1、第2、第3、第5、第7、第9、第11天内加德纳菌在阴道内的定植情况,以及阴道细菌结构和细菌生物多样性,并通过显微镜观察阴道组织的病理改变,比较3种造模方法的差异。另采用阳性药物(定君生,成分为德氏乳杆菌)对模型进行验证。结果采用链霉素联合抗链霉素加德纳菌能诱导形成典型的阴道菌群紊乱模型,造模后可见加德纳菌在阴道内的定植增多,优势菌以铜绿假单胞菌为主,且阴道细菌生物多样性增加,阴道组织可见明显的炎症浸润和上皮细胞坏死等病理改变。定君生能显著减少加德纳菌在阴道内的定植,减少阴道内的细菌生物多样性,并能显著改善阴道组织病理变化。结论采用链霉素联合抗链霉素加德纳菌诱导的小鼠阴道菌群紊乱模型方法更佳,模型的细菌学和组织病理学改变,以及对药物的反应性与临床有一定相似,具有临床应用价值。

Objective To establish a mouse model of vaginal flora disorder. Methods Estrogen treated C57 BL/6 mice were used to induce the mouse model of vaginal flora disorder by different ways(Streptomycin 50 μg/mouse;Gardnerella 50 μL/mouse;Streptomycin 50 μg/mouse+anti-Streptomycin Gardnerella 50 μL/mouse, 70 mice for each group). High-throughput sequencing of 16 S rDNA amplicons was used to observe the colonization of Gardnerella in vagina, and the bacterial structure and biodiversity were detected on the first, second, third, fifth, seventh, ninth and eleventh days after modeling. Then the pathological changes of vaginal tissue were observed under microscope, and the differences among the three methods were compared. In addition, positive drugs(Dingjunsheng, viable preparation of Lactobacillus) were used to verify the model. Results Streptomycin combined with anti-Streptomycin Gardnerella induced the formation of a typical vaginal flora disorder model. After the establishment of the model, the colonization of Gardnerella in vagina increased;the biodiversity of vaginal bacteria increased, with Pseudomonas aeruginosa being predominant;the pathological changes such as inflammatory infiltration and epithelial cell death were found in the vagina. Dingjunsheng significantly reduced the colonization of Gardnerella and bacterial biodiversity in vagina, and significantly improved the pathological changes of vagina. Conclusion The method of Streptomycin combined with Gardnerella is a better way to induce mouse model of vaginal flora disorder since the bacteriological and histopathological changes of the model, as well as the response to drugs are similar to clinical ones.