Effects Of Butyl Alcohol Extract Of Baitouweng Decoction On Virulence Factors Of Candida Albicans Isolated Form Vulvovaginal Candidiasis

Objective:To investigate the effect of Butyl alcohol extract of Bai Tou Weng decoction on virulence factors of Candida albicans.Methods:We use different polarity reagent to extract Bai Tou Weng decoction and to detect activity against C. albicans. XTT assay and agar plate assay were used to evaluate the effect of Butyl alcohol extract of Bai Tou Weng decoction(BAEB) against adhesion of C.albicans to 96-well plates and polyester catheters. SEM was applied to observe the morphological changes of C. albicans in adhesion at different times;The water-hydrocarbon two phase assay was applied to measure the cell surface hydrophobicity( CSH) of C. albicans; Test tube assay was utilized to evaluate flocculation Inverted microscope was used to find the best p H for morphological changing,fluorescence microscope and scanning electron microscope(SEM)were applied to inspect the morphological change of C.albicans treated by BAEB.Solid plate was utilized to the colonies forms.Sub-solid plate was used to evaluate to the hypha smite.Microdilution method and time-kill experiment were adopted to observe the activity against C. albicans CFU,Crystal violet staining method was used to determine the biomass of the biofilm, Scanning electron microscopy(SEM) was applied to observe the morphological changes of the biofilm.Confocal laser scanning microscopy(CLSM) was applied to determine the thickness of the biofilm. q RT-PCR was applied to evaluate expressions of virulence factors related genes.In vivo experiments to determine the treatment effect of BAEB to VVC?Results:(1) MIC of BAEB was within 64~256?g/m L,which was better than other extracts.(2) 512,1024 ?g/mL BAEB could inhibit the adherence of C. albicans to96-well plates and polyester catheters.(3) Hyphal grow well in p H8.0,512,1024 ?g/m L BAEB could inhibit hyphal development.(4) 1024?g/m L BAEB could inhibit the formation of C.albicans biofilm formating,Inverted microscope,fluorescence microscope and SEM,further confilmed intervation effect on morphological change.(5) Most of genes about virulence factors were up-regulated and down-regulated at different folds especial in 512 and 1024 ?g/m L group.(6) BAEB could effectively reduce the CFU in Vaginal lavage fluid,and suppress the hyphal around the Epithelial cells in mice Vaginal lavage fluid. HE staining shows BAEB can alleviate the inflammation in mice vagina.Conclusion: BAEB could inhibit adhesion, hypha formation, hydrophobicity and biofilm formation of C.albicans at some extent..When infected with VVC,BAEB showed therapeutic effect.