外阴阴道假丝酵母菌病模型大鼠阴道局部免疫调节的变化

Changes of local vaginal immune regulation in rats infected with vulvovaginal candidiasis

目的通过建立外阴阴道假丝酵母菌病(VVC)大鼠模型,研究VVC感染的疾病过程及不同治疗后阴道上皮的免疫修复情况以及阴道上皮细胞超微结构的变化。方法使用雌性SD大鼠建立VVC模型,并分别予不处理(模型对照组)、制霉菌素(制霉菌素组)治疗、中成药康妇消炎栓治疗(康妇消炎栓组),每组6只。对阴道上皮行透射电镜观察和免疫组化染色,观察上皮细胞超微结构的变化以及细胞因子γ-干扰素(IFN-γ)、白细胞介素(IL)4、IL-17、IgG在上皮细胞中的表达情况。结果模型对照组大鼠的病原体转阴率为0,制霉菌素组转阴率为6/6,康妇消炎栓组为5/6,3组比较,差异有统计学意义(P=0.015)。透射电镜下可观察到VVC感染后的阴道上皮细胞出现明显的结构破坏,而制霉菌素、康妇消炎栓治疗后的上皮细胞结构基本恢复至正常状态。免疫组化染色结果显示,康妇消炎栓组4种细胞因子中发挥免疫保护作用的IFN-γ、IgG表达升高,分别与正常对照组、模型对照组比较,差异有统计学意义(P<0.05),与制霉菌素组比较,差异无统计学意义(P>0.05);制霉菌素组IL-17的表达增高,而康妇消炎栓组IL-17的表达下降,两组比较,差异有统计学意义(P<0.05)。结论VVC感染后阴道上皮细胞的超微结构会发生明显的损伤变化,阴道局部免疫状态失调,抗真菌药物制霉菌素对于VVC有良好的治疗作用,可显著修复VVC感染后受损的阴道上皮结构,并可加强阴道上皮局部的免疫保护作用;中成药康妇消炎栓表现出与制霉菌素相似的治疗作用。

Objective To study the disease process of vulvovaginal candidiasis (VVC) infection in rat model of VVC, and to study the immuno-repairing effect of different treatments on vaginal epithelium and the ultra-structural changes of vaginal epithelial cells. Methods The VVC model of female rats were established. After successful modeling, the rats were treated with no treatment (model control group), nystatin and Kangfu Xiaoyan suppository. The vaginal epithelium was observed by transmission electron microscopy and immunohistochemical staining. The ultra-structural changes of epithelial cells and the expression of cytokines interferon γ(IFN-γ), interleukin (IL) 4, IL-17 and IgG in epithelial cells were observed and analyzed statistically. Results The negative conversion rate of model control group was 0, and that of nystatin group was 6/6, and that of Kangfu Xiaoyan suppository group was 5/6;significant difference existed between nystatin, Kangfu Xiaoyan suppository group and model control group (P<0.05). The ultrastructures of vaginal epithelial cells were damaged obviously after VVC infection, and the ultrastructures were repaired by nystatin and Kangfu Xiaoyan suppository under transmission electron microscope. Immunohistochemical staining showed, the expressions of IFN-γ and IgG in the four cytokines which played a protective role increased after Kangfu Xiaoyan suppository treatment, significantly different from that of model control group (P<0.05), but there were no significant differences of the IFN-γ and IgG expression between Kangfu Xiaoyan suppository group and those of nystatin group (P>0.05);the expression of IL-17 was increased after nystatin treatment, but decreased after Kangfu Xiaoyan suppository treatment, and the difference between the two groups had statistical significance (P<0.05). Conclusions The ultrastructure of vaginal epithelial cells after VVC infection could be damaged obviously, the local immune state is disordered, and the antifungal drug nystatin has a good therapeutic effect on VVC, it could significantly repair the damaged vaginal epithelium structure after VVC infection and strengthen the protective immune function of vaginal epithelium. Kangfu Xiaoyan suppository, one of Chinese medicine, has similar therapeutic effect with nystatin.