JieZeNO.1 Modify The Adhesion Of Vaginal Epithelial Cells-candida Albicans And The Expression Of Innate Immune Factors TLR4/NF-κB

Objective:Observing the effect of JieZe NO.1 on the adhesion between vaginal epithelial cells and Candida albicans cells; Demonstrating the possible mechinsm of the drug to treat Candidal albicans Vaginitis.Methods:There are five groups: model control, 12.5mg/ml JieZe NO.1 control, 5umol/L bererine control, 3mg/ml mannan control, 5mmol/L periodic control. In order to demonstrate and better unserstand the anti-Candida activity of the probiotic microorganisms in an in vitro model simulating vaginal candidiasis, a human vaginal epithelial cell lines(VK2/E6E7) was infected with C. albicans(ATCC90028) after challenged by medcines. At proper time points, the adheresion will be tested by Flow Cytometry.Results:1. The impact of drugs on cells.The impact of drugs on cells were studied by MTT assey. The IC50 of JieZe NO.1 on VK2/E6E7 was 100.8279mg/ml and the IC20 was 36.0579mg/ml; The IC50 of bererine on VK2/E6E7 was 21.2164umol/L and the IC20 was 10.696umol/L。2. The effects of JieZe NO.1 on the ability of vaginal epithelial cells to bind Candida albican cells.The adhesion were studied by flow cytometric analysis. And the adhesion of JieZe NO.1 control and mannose control produced a significant decrease in the number of vaginal epithelial cells that bound FITC-positive Candida compared with model control (11.05+0.3897% and 7.2163+0.3667% VS 24.5767+1.575%), p<0.05; On the contrary, periodic control produced had no effect on adherence between epithelial cells and Candida cells(22.7767+2.3034%), p>0.05.Conclusion:JieZe NO.1 produced a significant decrease the adhesion between VK2/E6E7 and Candida Albicans, which demonstrated that the drug can inhance the ability of VK2/E6E7 against Candida Albicans. Objective:Observing the effect of JieZe NO.1 on the expression of TLR4 and NF-κB after stimulated by Candida cells. To demonstrate the effects of this drug on partly molecular mechnism of innate immune responses about vaginal epithelial cells against Candida Albicans.Methods:There are six groups: normal control, model control, 12.5mg/ml JieZe NO.1 control, 5umol/L bererine control, 3mg/ml mannan control, 5mmol/L periodic control. In order to demonstrate and better unserstand the anti-Candida activity of the probiotic microorganisms in an in vitro model simulating vaginal candidiasis, a human vaginal epithelial cell lines(VK2/E6E7) was infected with C. albicans(ATCC90028) befor challenged by medcines. The time of co-incubation between host cells and C albican will be tested by immunofluorescent technique; and the expression of TLR4 and NF-κB by VK2/E6E7 will be tested by Western-blotting.Results:1. The time table of NF-κB in VK2/E7E7 activated by Candida cells. The time table was obversed by Immunostaining for Confocal microscopy. We found that the expression of TLR4 in the cytoplasma and NF-κB in the nucleus of VK2/E7E7 reached the peak level at 2h, and remained high level at 12h poststimulation, whereas we detected the expression of TKR4 and NF-κB after 12 hrs stimulating by Candida cells. 2. The expression of TLR4 and NF-κB in VK2/E6E7.The expression was detected by Western-blotting assey. The model control produced a increase in expression of TLR4 and NF-κB compared with normal control(P<0.01);. The JieZe NO.1, Bererine control and periodic control produced a decrease in expression of TLR4 and NF-κB compared with model control(P<0.01). The mannan control produced a slight decrease in expression of TLR4(P>0.05), and a increase in expression of NF-κB compared with model control(P<0.01).Conclusion:The data demonstrate that JieZe NO.1 downregulated the expression of TLR4 and inhibited the activity of NF-κB after VK2/E6E7 infected by conidia of Candida Albicans, which may downregulate the inflammtory cytokines, which can releave acute inflammtory symptom.