Study On Anti-candida Albicans Activity Of Sapindus Saponins In Vitro And In Vivo And Its Anti-biofilm Activity

Peel of Sapindus mukorossi Gaertn.contains abundant natural active ingredients saponins.Previous studies have shown that Sapindus saponins had antimicrobial,anti-inflammatory,whitening,antitumor and other activities.In this study,water extract fermentation liquid from peel of Sapindus riched in saponins was chosen and evaluated its antifungal activity and anti-biofilm effect on Candida albican which was a pathogenic fungus of vaginitis.This expanded utilization of Sapindus in the field of medicine.The main research contents and results were as follows:1.Active components analysis of Sapindus saponins.UPLC-TOF-MS was used to analyze the components and results showed that water extract fermentation liquid from peel of Sapindus mainly contained triterpenoid saponins and sesquiterpene glycosides.The oleanolic acid was used as the standard for the quantitative analysis of saponins.The total content of saponins was 117.12 mg/mL and the purity was 52.03%.2.Antifungal effect of Sapindus saponins on floating Candida albicans ATCC 10231.The minimum inhibitory concentration of Sapindus saponins to C.albicans was 0.08 mg/mL(calculated by saponins,the same below),and the minimum fungicidal concentration was about 0.32 mg/mL.In addition,the inhibitory rate could reach over 93.07% in a short time(4 h)at the concentration of 1 mg/mL.Subsequently,XTT colorimetric quantitative method was used to explore the effects of Sapindus saponins on biofilm of C.albicans.It was observed that the biofilm formed in the early stage could be inhibited at a low concentration(0.16 mg/mL),and mature biofilm could be removed at a relatively high concentration(0.64 mg/mL)compared with the positive control(0.64 mg/mL fluconazole).Further research showed that Sapindus saponins could reduce the adhesiveness and hydrophobicity of the surface of C.albicans.Confocal laser scanning microscopy,scanning electron microscope and inverted fluorescence electron microscope were utilized to observe the C.albicans biofilm and results showed that Sapindus saponins effectively reduced density and thickness of biofilm formation,made the cell surface wrinkled and uneven,and inhibited aggregation and mycelium transformation of C.albicans,respectively.The effect of Sapindus saponins on the expression of related genes was further studied by qRT-PCR.Results showed that Sapindus saponins would down-regulate the expression level of related ECE1?ALS3?and HWP1,and up-regulate the expression level of NRG1.Therefore,Sapindus saponins could inhibit the growth of biofilm of C.albicans mainly by reducing adhesion and mycelial transformation.3.The effect of Sapindus saponins on vaginitis in mice with C.albicans.In a mice model of vaginitis model,effects of Sapindus saponins on histopathology,fungal quantity,inflammatory factors and lactate dehydrogenase(LDH)were observed and analyzed after the treatment.Results showed that the number of fungi in Sapindus saponins treated group decreased from 3.57 log CFU/100 ?L to 1.84 log CFU/100 ?L after 6 d.Meanwhile,the levels of IL-1 ?,IL-6,IL-8 and LDH in Sapindus saponins treated group was lower than those in the untreated group.Therefore,Sapindus saponins would reduce the infiltration of neutrophils on the surface of vaginal epithelium.In addition,the influence of Sapindus saponins was explored on dysfunctional flora through the high-throughput sequencing of vaginal flora in mice.Compared with the model group,the proportion of bacteria in the two dominant groups(Proteobacteria and Firmicutes)in vaginal flora of mice was changed.The proportion was similar to that in the blank group and bacterial species were increased after the treatment of Sapindus saponins.In particular,proportion of Stenotrophomonas and Lactobacillus was increased after the Sapindus saponins treatment,which indicated that Sapindus saponins could restore some bacteria to normal level to some extent.To summarize,Sapindus saponins would inhibit C.albicans and its biofilm,improve the symptoms of candidal vaginitis,and make the vaginal flora tend to be normal.The results of this study provided evidence to support the treatment of vaginitis and the medical aspect of Sapindus.