Association Between Vaginal Micro-environmental Factors And Vaginal Floras In Different Cervical Lesions

ObjectiveCervical cancer is a complex disease infected by multi-factors and progressing in continuous steps.The relationship between vaginal micro-ecology and cervical cancer has arisen much concern nowadays.Present researches have shown that abnormality of vaginal micro-environment factors could increase risks of vaginitis and HPV infection.However,relationships between vaginal micro-environment factors and vaginal floras is not clear,as well as relationships between vaginal micro-environment factors and cervical carcinogenesis has not been reported.Subjects with normal cervix,cervical intraepithelial neoplasia and cervical cancer were enrolled in this study.Vaginal floras were taken as the entry point.Based on analyzing the association between vaginal micro-environment factors and vaginal floras and cervical carcinogenesis,the association and interaction between micro-environment factors and floras in cervical carcinogenesis was explored further.This study aimed to provide scientific basis for establishing early warning model and prevention strategy of cervical cancer.Methods290 women diagnosed with normal cervix?NC?,97 patients with low grade cervical intraepithelial neoplasia?CIN 1?,107 patients with high grade cervical intraepithelial neoplasia?CIN 2/3?and 16 patients with cervical squamous cell carcinoma?SCC?were enrolled in this study from January to June,2018 in gynecologic and obstetrics department of Second Hospital of Shanxi Medical University according to the inclusive and exclusive criteria strictly.Information related to cervical lesions was collected using a structured questionnaire,moreover,vaginal secretions and cervical exfoliated cells were collected from all participants.HPV infection was detected by flow-through hybridization.Vaginal H₂O₂,neuraminidase and leukocyte esterase were evaluated by Aerobic Vaginitis?AV?/Bacterial Vaginal Disease?BV?Five Joint Test Kit,in addition,cleanness and PH of vagina were detected.In the 16s rDNA sequencing phase,subjects who used antibiotics within 2 weeks or underwent vaginal medication/douche within 3 days were excluded.After matching by age,there were 22 women with NC,41 with CIN and 10 with SCC.Database was established using Epidata 3.1 software.The Chi-square test,logistic regression and Wilcoxon rank sum test were performed by SPSS22.0 software and GraphPad Prism7.0 software.Diversity and composition of vaginal floras and significant species analysis were performed after sequencing.Results1.Association between vaginal micro-environment factors and cervical carcinogenesis:The abnormal rates of PH and H₂O₂ were significantly different between NC,CIN and SCC groups?P<0.05?.Abnormality of PH?OR=1.78,95%CI:1.052-3.012?and H₂O₂?OR=1.94,95%CI:1.207-3.117?could increase risk of CIN.The abnormal rate of PH(?²_trend=6.530,P=0.011)and H₂O₂(?²_trend=8.249,P=0.004)gradually increased with the aggravation of cervical lesions.There were no significant association between cervical carcinogenesis and vagina cleanness,neuraminidase and leukocyte esterase.2.Association between vaginal PH,cleanness,H₂O₂ and vaginal floras:The dominant bacteria was Lactobacillus in women with normal PH,vagina cleanness and H₂O₂ with a proportion of 98.37%,59.60%and 95.70%respectively,while the top two bacteria in women with abnormal PH,vagina cleanness and H₂O₂ were Lactobacillus?47.27%,44.64%,45.20%respectively?and Gardnerella?17.81%,29.13%,18.81%respectively?.The characteristic bacteria was Lactobacillus among women with normal PH and H₂O₂,while Gardnerella and Atopobium in women with abnormal PH and Gardnerella in women with abnormal H₂O₂.Characteristic bacteria was not found in women with normal or abnormal vagina cleanness.Diversity of vaginal floras was significantly higher in women with abnormal PH,vagina cleanness and H₂O₂?P<0.05?.3.Association between neuraminidase and leukocyte esterase and vaginal floras:The dominant bacteria was Lactobacillus among women with normal neuraminidase and leukocyte esterase with a proportion of 59.99%and 62.19%respectively,while Gardnerella in women with abnormal neuraminidase and leukocyte esterase,accounting for 36.23%and 30.23%respectively.The characteristic bacteria were Bacilli and Firmicutes in women with normal leukocyte esterase,while Peptoniphilus,Prevotella and Porphyromonas among women with abnormal leukocyte esterase.In addition,characteristic bacteria in women with both normal and abnormal neuraminidase was not found.Diversity of vaginal floras in women with abnormal leukocyte esterase was significantly higher?P<0.05?.4.Interaction and association between micro-environment factors and vaginal floras in cervical carcinogenesis:Interaction analysis of micro-environment factors related to cervical carcinogenesis and vaginal floras in cervical carcinogens is showed as follows.In NC group,Lactobacillus was the dominant bacteria in women with both normal and abnormal PH?99.20%and 69.25%?as well as H₂O₂?92.96%and63.25%?;the characteristic bacteria was Lactobacillus in women with normal PH and H₂O₂,while Dialster in women with abnormal PH and Staphylococcus and Lactococcus among women with abnormal H₂O₂;diversity of vaginal floras in women with abnormal PH and H₂O₂ was not significantly higher?P>0.05?.In CIN group,Lactobacillus was the dominant bacteria in women with both normal and abnormal PH?97.00%and 52.45%?as well as H₂O₂?92.15%and 51.03%?;the characteristic bacteria was Lactobacillus in women with normal PH and H₂O₂,while Anaerococcus,Dialster,Parvimonas and Aerococcus in women with abnormal PH and Gardnerella,Atopobium,Mobiluncus,Gemella,Dialster,Anaerococcus and Parvimonas among women with abnormal H₂O₂;diversity of vaginal floras in women with abnormal PH and H₂O₂ were significantly higher?P<0.05?.In SCC group,Lactobacillus was the dominant bacteria in women with normal PH?96.80%?;results of H₂O₂ in all SCC subjects were abnormality;Peptoniphilus was the dominant bacteria in women with abnormal PH?22.68%?and H₂O₂?22.76%?;no characteristic bacteria was found in women with abnormal or normal PH as well as abnormal H₂O₂;diversity of vaginal floras in women with abnormal PH and H₂O₂ was not significantly higher?P>0.05?.The additive interaction model showed that there was an additive interaction between the abnormal PH,abnormal H₂O₂ and increase of diversity of the vaginal flora in CIN,however,no similar result was found in SCC.Conclusion1.Abnormal vaginal micro-environment factors can increase the risk of cervical intraepithelial neoplasia,suggesting that strengthening the detection of vaginal micro-environmentfactorsandmaintainingthestabilityofvaginal micro-environments is of great significance for the prevention of cervical intraepithelial neoplasia and the blocking of the process of cervical cancer.2.The diversity of vaginal floras increased among women with abnormal vaginal micro-environment factors,meanwhile,dominant bacteria and characteristic bacteria was found in different status of vaginal micro-environment factors,suggesting the detection of vaginal micro-environmental factors can provide an important reference for the evaluation of vaginal floras change and regulation of vaginal floras.3.The abnormality of vaginal microenvironment factors was related to the increase of vaginal flora diversity and the change of dominant bacteria and characteristic bacteria in different cervical lesions,in addition,there were positive additive interactions between the abnormal vaginal micro-environment factors and the increase of vaginal floras diversity in CIN group,suggesting that the synchronous detection of vaginal micro-environment factors and vaginal floras is of great significance for the improvement of vaginal micro-environment,early detection of cervical cancer and blocking the process of cervical cancer.