Correlation Analysis Between The Vaginal Microecology And Immune Status And The Cervical High-Risk Human Papillomavirus Infection

Objective:To study and analyze the vaginal microecology of patients with High risk Human papillomavirus（HR-HPV）infection,and the cytokine macrophage inflammatory protein 3α（MIP-3α）and interleukin-10（Interleukin-10,IL-10）,Interleukin-1β（IL-1β）,Interleukin-8（IL-8）,Interleukin-6（IL-6）,Macrophage Colony Stimulating Factor,GM-CSF),Tumor Necrosis Factor（TNF）and monocyte chemoattractant protien-1（MCP-1）,and to explore the effects of vaginal immune function and vaginal microecology on cervical high-risk Human Papillomavirus infection and cervical lesions,in order to promote the negative transformation of human papillomavirus（HPV）infection and cervical lesions.Methods:（1）In this study,the patients with HPV infection and the healthy people in the gynecological clinic of Liuzhou People’s Hospital were selected.According to the inclusion and exclusion criteria drawn up in this study,the patients diagnosed as high-risk HPV were included.According to the results of HPV and Thinprep cytologic test（TCT）,there were 30 cases in HPV+and TCT+groups.30 cases in HPV+and TCT-groups were the experimental group and 30 healthy people with negative HPV and TCT tests in the same period were the control group.（According to the diagnosis and treatment plan of HPV infection,two subjects whose final pathological results were the cervical malignant tumor and one subject who used vaginal medication before sampling were excluded,so finally they were divided into HPV+and TCT+groups,31 cases in HPV+and TCT-groups,30 cases in HPV+and TCT-groups and 27 cases in HPV-and TCT-groups,with a total of 88 cases.）,the analysis and statistics process divided HPV infected people into two groups according to whether they were single HPV infection or not（35 cases in single HPV+group and 26 cases in multiple HPV+groups,a total of 61 cases）.（2）Take vaginal exfoliated cells from each group for microecological examination,and analyze the differences and correlation between groups.（3）The concentrations of cytokines IL-6,GM-CSF,TNF,MCP-1,MIP-3α,IL-10,IL-1β and IL-8 in vaginal lavage fluid of each group were detected by ELISA.It was found that the detection rates of cytokines IL-6,GM-CSF,TNF and MCP-1 were low,and finally the cytokines MIP-3α,IL-10,IL-1β and IL-8 were selected.（4）Statistical analysis was carried out by SPSS26.0 statistical software,in which the counting data were expressed by rate（%）,and statistical analysis was carried out by chi-square test or Fisher exact probability method;The normal distribution in the measurement data is expressed by the mean standard deviation（χ±s）,and the statistical analysis is carried out by T test,and the comparison between groups is carried out by analysis of variance.Non-normal distribution is represented by median（M）and quartile（IQR）,and nonparametric test is used for statistical analysis.Mann-Whitney rank sum test is used for comparison between two independent samples,and Kruskal-Wallis rank sum test is used for comparison among multiple independent samples.P<0.05 is statistically significant.Results:（1）In the HPV positive group,the positive rate of leukocytes,the infection rate of candida vaginitis（VVC）,the normal rate of lactobacillus,the positive rate of leukocyte esterase and the abnormal rate of flora diversity were all lower than those in the HPV negative group,but there was no statistical difference between the two groups（P>0.05）;The infection rate of Bacterial vaginosis（BV）,the normal rate of H₂O₂,the decrease rate of flora density,the positive rate of sialidase and the abnormal rate of Nugent score in HPV positive group were higher than those in HPV negative group,but there was no statistical difference between the two groups（P>0.05）.The abnormal rate of PH value in HPV positive group was higher than that in HPV negative group,and the difference between groups was statistically significant（P<0.05）.Trichomonas,mycoplasma and chlamydia were negative in all groups.（2）In single HPV+group and multiple HPV+groups,the positive rate of leukocytes,BV infection rate,VVC infection rate,abnormal rate of PH value,normal rate of lactobacillus,decreased rate of flora density,negative rate of H₂O₂ and decreased rate of flora diversity were all higher than those in multiple HPV+groups,but there was no statistical difference between the groups（P>0.05）.Compared with other HPV+groups,the positive rate of sialidase,the increase rate of flora density and the abnormal rate of Nugent score decreased,but there was no statistical difference between the two groups（P>0.05）.（3）Respectively in HPV+,TCT+、HPV+,TCT-and HPV-and TCT-groups,the median concentrations of IL-1βwere 30.32 pg/ml,8.62pg/ml and 8.04 pg/ml;the median concentration of MIP-3α was 14.43pg/ml,13,67 pg/ml and 17.19pg/ml;the median concentrations of IL-8 were 375.60pg/ml,57.29pg/ml and 17.19pg/ml;the median concentrations of IL-10 were 96.78pg/ml,98.92pg/ml and 59.92pg/ml.The median concentrations of IL-1β,IL-8 and IL-10 in HPV positive group were higher than those in HPV negative group,and the median concentrations of IL-1β and IL-8 in HPV positive group which caused cytological changes were higher than those in HPV positive group only,with statistical significance（P<0.05）.The median concentration of IL-10 in the HPV-positive group that caused cytological changes was lower than that in the pure HPV-positive group,and there was a statistical difference between the two groups（P<0.01）.The median concentration of MIP-3α in HPV-positive group decreased compared with HPV-negative group,and the median concentration of HPV-positive group which caused cytological changes increased compared with HPV-positive group,and there was no statistical difference between the two groups（P>0.05）.（4）Respectively in single HPV+group and multiple HPV+groups,the median concentration of IL-1β was 12.27pg/ml and 78.27pg/ml;The median concentration of MIP-3α was 15.94pg/ml and 8.00 pg/ml;The median concentration of IL-8 was 324.52pg/ml and 724.79pg/ml;The median concentration of IL-10 was 95.93pg/ml and 106.64pg/ml.The median concentration of IL-1β and IL-8 in single HPV positive group was lower than that in multiple HPV positive groups,and the difference was statistically significant（P<0.05）.The median concentration of MIP-3α increased in single HPV positive group compared with multiple HPV positive groups,and there was no significant difference between the two groups（P>0.05）.The median concentration of IL-10 in single HPV positive group decreased compared with that in multiple HPV positive groups,and there was no significant difference between the two groups（P>0.05）.Conclusion:（1）HPV infection may be related to vaginal microecological imbalance;Among them,PH>4.5 may be related to the occurrence of HPV infection.However,PH>4.5 can not be proved to be related to multiple HPV infections.（2）This study suggests that the infection of BV,VVC,trichomonas,mycoplasma and chlamydia can not prove to be related to HPV infection.（3）Vaginal immune dysfunction may be related to cervical high-risk HPV infection;Among them,high-risk HPV infection in cervix may be related to the increase of IL-1β,IL-8 and IL-10 concentrations.（4）HPV multiple infection may be related to the increase of IL-1β and IL-8 concentration;The increase of IL-10 concentration cannot be proved to be related to HPV multiple infection.