To Explore The Effect Of Blaps Rynchopetera Fairmaire Crude Extract On Autophagy Level In Rats Aerobic Vaginitis

【Objective】To study the effects of the crude extract of Blapsrynchopetera Fairmaire(BRF)on the expression of autophagy marker proteins Beclin1 and LC3 B in the vagina of rats with aerobic vaginitis,the mechanism of crude extract of BRF on the treatment of aerobe vaginitis(AV)was preliminarily explored from the viewpointof autophagy.【Method】1.Extraction of antibacterial components: 60% ethanol soaks the adults BRF,The soaked extract is extracted with petroleum ether,chloroform,ethyl acetate and n-butanol to obtain the freeze-dried crude extract of four polar parts.Detect the minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of the three common pathogens(Staphylococcus aureus,Escherichia coli,Streptococcus agalactiae)of the three common AV,and select the most effective crude antibacterial extract for Animal experiment.2.Animal experiment: 50 mature female SD rats,normal group(without any treatment),model group,antibiotic group,extract low-concentration,medium-concentration and high-concentration groups(S.aureus,E.coli and S.agalactiae mixed liquid were successfully simulated in vagina).The model group was given cream matrix,the antibiotic group was given vaginal cream with clindamycin phosphate(containing 20mg/g),and the low,medium,and high concentration groups were given the crude extract of BRF.They contained 100 mg/g,200 mg/g,and 300mg/g homemade cream with ethyl acetate.After 14 days of treatment,blood were collected from the abdominal aorta and the rats were sacrificed.3.Data collection:(1)Measure the modified Donders score and p H value of vaginal secretions of each group of rats before and after treatment,(2)HE staining of vaginal tissues,(3)immunohistochemical(IHC)detection of each group rats before and after treatment of vaginal tissues Beclin1 and LC3 B expression,(4)Western blot detection between groups before and after treatment of vaginal tissues of rats Beclin1 and LC3B protein expression,(5)ELISA detection between groups before and after treatment of vaginal tissues of rats Beclin1 and LC3 B expression.【Results】1.The crude extracts of BRF methyl petroleum ether and n-butanol did not show in vitro antibacterial activity against E.coli.The MIC of crude chloroform and ethyl acetate extracts against E.coli was 2.50mg/ml,1.25mg/ml,the MBC was 2.50mg/ml,1.25mg/ml;All the crude extracts showed different degrees of antibacterial and bactericidal activities against S.aureus and S.agalactiae,and ethyl acetate was the best crude extract screened in this experiment.2.After the treatment of AV model rats,the modified Donders score,p H value and vaginal histopathology score of vaginal secretions in the model group and the crude extract low concentration group were not statistically different from those in the normal group.The vaginal histopathological scores of other groups were reduced to different degrees,and the three indicators were reduced to different degrees.Among them,the high concentration group of crude extract was significantly lower than the concentration group of crude extract.3.After treatment,the protein expressions of Beclin1 and LC3 B in the model group and the low concentration crude extract group showed no statistically significant difference compared with the normal group by IHC,Western blot and ELISA,while the protein expressions in the antibiotic group,medium and high concentration crude extract group decreased to varying degrees.【Conclusions】1.The ethyl acetate part of BRFcrude extract has antibacterial activity against S.aureus,E.coli and S.agalactiaein vitro,which werecommon pathogens of AV;2.The ethyl acetate part of BRFcrude extracthad a therapeutic effect on AV in rats.After treatedwith medium and high concentration ethanol extract,the modified Donders score,p H valueof vaginal secretions and vaginal histomathological score in rats with AV were decreased,and the efficacy was dose-dependent.3.Theautophagy level in the vaginal tissues of the rats AV model was significantly increased.The crude extract of BRF ethyl acetate achieved the effect of treating AV throughinhibit the excessive activationof autophagy.