益生植物乳杆菌抑制白色念珠菌的分子机制

Candida albicans is a common opportunistic pathogenic fungus that can cause many diseases such as vulvovaginal candidiasis(VVC) in women. With the wide application of antifungal drugs, drug resistance has become a clinical treatment problem. The use of human normal flora Lactobacillus for the treatment of VVC has made some progresses, but the specific molecular mechanism research is relatively less. Our previous study has found that RNA in extracellular vesicles(EVs) secreted by probiotic Lactobacillus plantarum can inhibit the growth of Candida albicans and induce apoptosis. This study intending to further clarify the specific mechanism of RNA in Lactobacillus plantarum EVs induce apoptosis of Candida albicans by utilizing cell biology and molecular biology techniques. Construct cell and animal VVC infection models to evaluate the treatment of VVC by RNA in EVs. Using second-generation sequencing technology to analysis the transcriptome sequence of RNA in EVs, and predict small interfering RNA which can induce apoptosis in Candida albicans. Applicate corresponding molecular biology methods to verify the specific function of small interfering RNA. To elucidate the molecular mechanism of Lactobacillus plantarum inhibiting the growth of Candida albicans, and provide new theoretical support and experimental basis for clinical treatment of VVC.

白色念珠菌是常见的机会性致病真菌，易引起女性外阴阴道念珠菌病（vulvovaginal candidiasis, VVC）等多种疾病。随着抗真菌药物的广泛应用，耐药现象已成为临床治疗难题。应用人体正常菌群乳杆菌治疗VVC已取得一定进展，但具体机制研究较少。我们前期发现，益生植物乳杆菌所分泌的细胞外囊泡(extracellular vesicles, EVs)内RNA可以抑制白色念珠菌生长，诱导其产生凋亡现象。本研究拟利用细胞生物学和分子生物学等技术，明确植物乳杆菌EVs内RNA诱导白色念珠菌产生凋亡现象的具体机制；构建VVC细胞和动物感染模型，评价应用EVs内RNA治疗VVC的效果；应用二代测序技术对EVs内RNA进行转录组测序，预测能够诱导白色念珠产生凋亡现象的小干扰RNA，验证小干扰RNA具体功能。阐明植物乳杆菌抑制白色念珠菌生长的分子机制，为临床治疗VVC提供新的理论支撑和实验依据。

随着抗生素及免疫抑制剂的广泛应用，真菌感染现象逐年增多，同时临床耐药菌株的数量也不断增加，应用有效且毒副作用小的抗真菌药物进行治疗具有重要的临床意义。乳杆菌是人体内的正常菌群之一，临床应用乳杆菌治疗VVC已取得一定成果，但具体分子机制研究较少。我们前期研究发现，益生植物乳杆菌分泌的细胞外囊泡内RNA 可以抑制白色念珠菌生长，诱导其产生凋亡现象。通过本研究拟进一步利用细胞生物学和分子生物学等技术，明确了植物乳杆菌EVs内RNA诱导白色念珠菌产生凋亡现象的具体机制；通过构建VVC细胞及动物感染模型，我们发现EVs内RNA具有良好治疗VVC的效果；同时，应用二代测序技术对植物乳杆菌EVs内RNA进行转录组测序分析，预测得到可能诱导白色念珠产生凋亡现象的siRNA26条，并对其中三条siRNA进行了功能验证。本项目的开展对阐明植物乳杆菌抑制白色念珠菌生长的分子机制，以及为日后临床治疗VVC提供新的理论支撑和实验依据。