| Objective:By observing the effects of "Compound Phellodendron Liquid "and positive control drugs on the pathological changes of vaginal tissue sections in BV rat model caused by Staphylococcus aureus and Escherichia coli mixed liquid infection,the expression levels of IL-1β,IL-2 and IL-10 in vaginal lavage fluid and NF-κBp65 in vaginal tissue.To investigate the curative effect and mechanism of compound Phellodendron liquid in the treatment of BV,and to provide experimental basis for clinical application of compound Phellodendron liquid in the treatment of BV patients in the future.Methods:1.Choose 8 weeks of SPF health,female,55 only virgin rats,adaptability to feed a week during this period,PBS solution irrigation vaginal injections into the vagina,removing the dirt,Continuous defouling for 6 days,and on the 1st,3rd and 5th day,0.2 mL of veterinary estradiol benzoate injection was injected for pseudo-estrangic cycle treatment,and the estrus status of rats was observed on day 6,randomly selected from 10 as blank group,the only remaining 45 rats by staphylococcus aureus and escherichia coli mixed microbial infection method building directly,After modeling,the rats were routinely fed for 4 days,and then 5 rats were randomly selected for sampling to judge whether modeling was successful.2.After successful modeling,the remaining model rats were divided into groups(compound Phellodendron liquid group,Honghe Fujie Lotion group,metronidazole vaginal gel group,model group),with 10 rats in each group,and treated for 7 consecutive days.3.Four days later,the drug was stopped,each rat lavage vaginal adequately with sterile PBS solution,each are vaginal lavage fluid retention,cervical vertebra to be put to death in the rat off the next day,take the vaginal tissue,macroscopic observation and pathological records,under the microscope after HE staining histomorphology changes,and the degree of damage to the vaginal tissue classification rating,The expression of NF-κBp65 protein was determined by immunohistochemistry,the expression level of NF-κBp65 pathway protein was detected by Western blotting,and the expression level of IL-1β,IL-2 and IL-10 in vaginal lavage fluid was detected by Elisa.Results:1.General condition of rats:rats in each group had good mental state and activity;The fur darkened only after injection of estradiol benzoate and returned to normal after 2-3 days.After modeling,vaginal secretion increased and decreased gradually during drug treatment.2.Vagina histopathology of rats:Direct visual observation:Kruskal-wallis test showed that,compared with the model group,the curative effect of compound Phellodendron liquid group and metronidazole vaginal gel group was significantly different(P<0.05),and the curative effect of Honghe Fujie Lotion group was significantly different(P<0.01).The results of vaginal tissue injury score showed that compared with blank group,the vaginal tissue injury score of model group was significantly higher(P<0.01).Compared with model group,vaginal tissue injury score in metronidazole vaginal gel group was significantly lower(P<0.01).There was no significant difference in the vaginal tissue injury score between the compound Phellodendron liquid group and the Honghe Fujie Lotion group(P<0.05).3.Immunohistochemical results showed that Compared with the empty group,the proportion of NF-κBp65 nuclear translocation positive cells in the vaginal tissue of the module rats increased(P<0.05),and the proportion of NF-κBp65 nuclear translocation positive cells in the vaginal tissue of the remaining rat groups decreased significantly(P<0.01).4.Expression of IL-1β in vaginal lavage fluid of rats;Compared with blank group,the concentration of IL-1β in vaginal lavage fluid of model group was significantly increased(P<0.01);Compared with model group,the concentration of IL-1β in Honghe Fujie Lotion group was significantly decreased(P<0.01);The concentration of IL-1β in vaginal lavage fluid of rats in compound Phellodendron liquid group was significantly decreased(P<0.05);The concentration of IL-1β in vaginal lavage fluid of metronidazole vaginal gel group was decreased(P>0.05).5.Expression of IL-2 in vaginal lavage fluid of rats:Compared with blank group,the concentration of IL-2 in vaginal lavage fluid of rats in model group was significantly increased,with extremely significant difference(P<0.01).Compared with model group,the concentration of IL-2 in vaginal lavage fluid of rats in other groups showed a decreasing trend(P>0.05).6.Expression of IL-10 in vaginal lavage fluid of rats:Compared with blank group,the concentration of IL-10 in vaginal lavage fluid of rats in model group showed a downward trend(P>0.05).Compared with model group,the concentration of IL-10 in vaginal lavage fluid of rats in each group had an increasing trend(P>0.05).7.NF-κBp65 in vaginal tissues of rats:Compared with blank group,the protein concentration of NF-κBp65 in vaginal tissues of rats in model group was significantly increased,with statistical significance(P<0.05);Compared with model group,the protein concentration of NF-KBp65 in vaginal tissues of rats in all groups had a downward trend(P>0.05).8.Correlation analysis:The concentration of NF-κBp65 protein was positively correlated with the expression of IL-1β,with correlation coefficient=0.490(P<0.01).The concentration of NF-κBp65 protein was positively correlated with the expression of IL-10,the correlation coefficient was 0.391,and the correlation was statistically significant(P<0.05).Conclusion:1.Compound Phellodendron Liquid can reduce the vaginal tissue damage score of BV model rats and reduce the inflammation of rat vaginal tissue,indicating that the compound Phellodendron liquid has a therapeutic effect on BV rats2.The concentration of NF-κBp65 protein in the WB detection model group was increased,and the NF-κBp65 nuclear translocation in the histation model group was exempted,indicating that there was abnormal activation of NF-κB during the pathogenesis of BV rats3.The pathogenesis of BV is related to the abnormal activation of NF-κB and the immune shift of Th1/Th2,and the mechanism of compound Phellodendron liquid to treat BV may be:by inhibiting the expression of pro-inflammatory factors such as IL1β and IL-2,promoting the expression of anti-inflammatory factors such as IL-10,inhibiting abnormal activation of NF-κB,restoring the immune imbalance of Th1/Th2 cells,reducing the inflammatory response,improving the vaginal microenvironment,and treating bacterial vaginosis. |
