The Effect Of Estrogen On The Colonization Of Escherichia Coli And Candida Albicans In Vaginal Epithelial Cells And Its Mechanism Of Action

Part Ⅰ:Influence of estrogen on cell behavior and function of vaginal epithelial cell line VK2/E6E7Estrogen is an important sex hormone in women,which is especial for the maintenance of vaginal ecological balance.At physiological circumstances,Estrogen promotes thickening of vaginal epithelium and increases intracellular glycogen content.Vaginal epithelial cells glycogen is divided into monosaccharides which are broken down by vaginal lactobacilli into lactic acid to maintain normal vaginal acidic environment(pH≤4.5)and pnhibit the growth of other pathogens.And how estrogen affects the colonization of pathogen in vaginal epithelial cell and its mechanism has not been reported in the literature.Objective:To clear and define influence of 17β-estradiol on cell behavior and function of VK2/E6E7.Methods:Cultured VK2/E6E7 cells in vitro,detected influence of 17β-estradiol and estrogen-receptor inhibitor ICI82780 on cell proliferation of VK2/E6E7 in the method of MTS;detected influence of 17β-estradiol and ICI82780 on cell apoptosis of VK2/E6E7 by FCM;separately detected influence of 17β-estradiol on cell migration and invasion ability through migration experiment(Transwell assay and scratch)and invasion experiment.Added ICI82780 to observe influence of 17β-estradiol on cell migration and invasion ability of VK2/E6E7 after estrogen receptor a(ERa)and estrogen receptor β(ERβ)was inhibited.Results:lOnM and 100nM 17β-estradiol could accelerate cell proliferation of VK2/E6E7,with salt concentration dependence,the acceleration of 100nM was stronger,but 1000nM 17β-estradiol inhibited cell proliferation of VK2/E6E7,influence of InM 17β-estradiol on cell proliferation was not obvious;estrogen-receptor inhibitor ICI82780 with any concentration had no obvious influence on cell proliferation of VK2/E6E7.lOnM,100nM and 1000nM 17p-estradiol reduced early apoptosis rate of VK2/E6E7,while increasing its late apoptosis rate,but it had no obvious influence on overall cell apoptosis rate;0.1μM,1μM and 10μM 17β-estradiol increased early apoptosis rate of VK2/E6E7,while reducing its late apoptosis rate,but it had no obvious influence on overall cell apoptosis rate.10nM and 100nM 17β-estradiol enhanced cell migration and invasion ability of VK2/E6E7,with dose dependence,acceleration of 100nM 17β-estradiol on cell migration and invasion of VK2/E6E7 was more obvious,1nM 17β-estradiol and 0.1%alcohol had no obvious acceleration on migration and invasion of VK2/E6E7,1000nM 17β-estradiol could inhibit cell migration and invasion of VK2/E6E7;ICI82780 could inhibit the acceleration of 17β-estradiol on cell migration and invasion of VK2/E6E7,among them,inhibition of 10μM ICI82780 was the most obvious.Conclusions:Estrogen can accelerate cell proliferation,migration and invasion of VK2/E6E7,except for cell apoptosis,acceleration of estrogen on cell proliferation,migration and invasion of VK2/E6E7 cell might be mainly realized through its combination with ERa and ERβ to regulate the expression of downstream signal paths.Part Ⅱ:Influence and functional mechanism of estrogen for colonization of Escherichia coli and Candida albicans on epithelial cell line VK2/E6E7Objective:To clear and define influence of 17β-estradiol for Escherichia coli(E.coli)and Candida albicans in vaginal epithelial cell colonization,and understand possible functional mechanism.Methods:Cultured VK2/E6E7 cells to explore appropriate time for adhesion and colonization of E.coli and Candida albicans in VK2/E6E7 cells in vitro;explored the most appropriate bacteria chroma for adhesion and colonization of E.coli and Candida albicans in VK2/E6E7 cells in vitro;compared the blank group and the solvent group to detect influence of 17β-estradiol with different concentrations for E.coli and Candida albicans in VK2/E6E7 adhesion and colonization;added estrogen receptor antagonist ICI82780 to inhibit estrogen receptors a(ERa)and estrogen receptors β(ERβ),observed influence of 17β-estradiol for E.coli and Candida albicans in VK2/E6E7 adhesion and colonization again.Results:3h after microbial vaccination,the amount of colonized Candida albicans and E.coli in VK2/E6E7 cells was the most;when the vaccination concentration was 1.5×107 CFU/ml Candida albicans and 2.0×109 CFU/ml E.coli,the amount of colonized Candida albicans and E.coli in VK2/E6E7 cells was the most;24h after the treatment of lOnM and 100nM 17β-estradiol,the amount of colonized Candida albicans and E.coli in VK2/E6E7 cells was the most,with statistical significance,but 1000nM 17β-estradiol could inhibit colonization of Candida albicans with small extent and without statistical significance;after the treatment of 10μM ICI82780,the acceleration of 17β-estradiol for colonization of Candida albicans and E.coli in VK2/E6E7 cells was obviously inhibited,with statistical significance.Conclusions:Estrogen can accelerate adhesion and colonization of Candida albicans and E.coli in VK2/E6E7 cells,the acceleration is mainly realized by the combination of ERa and ERβ to regulate the expression of downstream signal paths,and ultimately increase the colonization ability of Candida albicans and E.coli.