低强度脉冲超声联合制霉菌素可协同抑制兔阴道白色念珠菌生物被膜感染  

Low-intensity pulsed ultrasound combined with nystatin treatment synergistically inhibits vaginal Candida albicans biofilm infection in rabbits

作　　者：谢梦瑶[1] 杨敏[1] 李欣[1] 杜永洪[1] XIE Mengyao;YANG Min;LI Xin;DU Yonghong(State Key Laboratory of Ultrasound in Medicine and Engineering,College of Biomedical Engineering,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学超声医学工程国家重点实验室//生物医学工程学院,重庆400016

出　　处：《南方医科大学学报》2025年第2期296-303,共8页Journal of Southern Medical University

基　　金：重庆市自然科学基金(CSTB2022NSCQ-MSX0124);重庆市教委科学技术研究计划项目(KJZD-K202400408)。

摘　　要：目的探索低强度脉冲超声(LIPUS)联合制霉菌素(NYS)对阴道白色念珠菌生物被膜感染的治疗效果。方法建立体外白色念珠菌生物被膜模型,同时将体外培养成熟的生物被膜用自动微移液管接种至20只雌性新西兰大白兔阴道内,以建立兔阴道白色念珠菌生物被膜感染模型,实验分为Control组、NYS组、US组和联合治疗组。体外实验采用XTT法检测生物被膜最低抑菌浓度(MIC),结晶紫染色定量生物被膜,激光共聚焦显微镜(CLSM)观察生物被膜活性,扫描电镜(SEM)观察形态学变化,DCFH-DA检测活性氧(ROS)。体内实验通过观察兔外阴症状,HE染色和透射电镜(TEM)观察治疗前后阴道组织病理结构变化。结果体外实验显示,联合治疗组MIC_(50)和MIC_(80)相比NYS组均降低2倍(P<0.05),此外,联合治疗组生物被膜清除率分别比NYS组和US组增加约26%和68%(P<0.001),同时联合治疗组生物被膜活性更低、结构破坏更严重,ROS产量更多。体内实验表明,与NYS组和US组比较,联合治疗组兔外阴症状与炎症浸润显著改善(P<0.05),阴道残留菌丝/菌株减少,上皮超微结构明显改善。结论LIPUS联合NYS对体内外白色念珠菌生物被膜有显著的协同抗真菌作用。Objective To explore the efficacy of low-intensity pulsed ultrasound(LIPUS)combined with nystatin for treatment of vaginal Candida albicans biofilm infection.Methods In vitro cultured Candida albicans biofilm were treated with LIPUS,nystatin,or both,and the minimum inhibitory concentration(MIC)of nystatin was determined.Crystal violet staining,confocal laser microscopy(CLSM)and scanning electron microscopy were used to quantify the biofilm and observe the activity and morphological changes of the biofilms;DCFH-DA was used to detect the changes in reactive oxygen species(ROS).Twenty female New Zealand White rabbits with vaginal inoculation of Candida albicans biofilm were randomized into 4 groups for treatment with normal saline,LIPUS,nystatin,or both LIPUS and nystatin.The changes in vulvar symptoms of the rabbits were observed,and the histopathological and ultrastructural changes of the vagina before and after treatment were observed using HE staining and transmission electron microscopy.Results In the combined treatment group,the MIC_(50) and MIC_(80) of nystatin in Candida albicans biofilms were both reduced by 50% compared with those in nystatin group,and the biofilm clearance rate increased by 26% and 68% compared with nystatin and LIPUS groups,respectively.Compared with nystatin and LIPUS treatment alone,the combined treatment produced stronger effects for inhibiting biofilm activity,causing structural disruption and promoting ROS production.In the rabbit models,the combined treatment more effectively improved vulvar symptoms and inflammatory infiltration,reduced residual vaginal hyphae/strains,and improved ultrastructure of the vaginal epithelium than LIPUS and nystatin treatment alone.Conclusion LIPUS combined with nystatin produces a significant synergistic antifungal effect against Candida albicans biofilm both in vitro and in vivo.

关 键 词：低强度脉冲超声 白色念珠菌 生物被膜 制霉菌素 阴道炎 

分 类 号：R711.31[医药卫生—妇产科学]