| Objective1.Clinical studyTo observe the effect of Youdujing vaginal suppository on HPV clearance and pathological prognosis of patients dignosed as human papillomavirus infection.2.In vitro studyTo observe the effect of Youdujing on the proliferation and apoptosis of human cervical immortalized cell H8 cells,and reveal its potential molecular mechanism.Methods1.Clinical studyA prospective controlled was designed in our trial.Patients dignosed as HPV infection were randomly divided into Youdujing vaginal suppository group and blank group.Patients in Youdujing vaginal suppository group were treated with suppository before going to bed every other day in non-menstrual period.Twelve times a month for a course,a total of three courses were needed.The blank group was observed for 3 months without invention.All patients should intercourse with condoms.At the end of the study,HPV clearance and cervical biopsy wre observed in both groups.2.In vitro studyH8 cells,which were transfected with HPV-16 gene-type and passed stably,were chosen to be model of our research.MTT assay was used to detect the proliferation of H8 cells.Apoptotic changes of the cells,invented by Youdujing for 24h,stained by Hoechst 33342 were observed by fluorescence microscope.Flow cytometry was used to detect the early apoptotic rate of H8 cells after Annexin V/PI double stained.Expressions of caspase-3 protein and PARP protein in H8 cells,which were treated with Youdujing and blank concentration for 24h,were tested by Western blot method.Results1.Clinical studyIn this study,the clearance rate of HPV was 77%,the total effective rate was 88%,while the blank rate was 27%after 3 months of intervention.There was significant difference between the experimental group and the blank observation group(F<0.05).There was no significant different between Youdujing vaginal suppository invention group on the points of Cervical biopsy.The points of Pathology confirmed difference between the two groups was statistically significant,while the test group(0.55 ± 0.10)was significantly lower than that of the blank control group(1.45 ± 0.82).It can be inferred that Youdujing vaginal suppository can reduce foci.There were 0 cases progressing,3 cases persisting,6 cases degrating of pathological change on experimental group;while 1 cases progressing,8 cases persisting,2 cases degrating on control group.There was no significant different between Youdujing vaginal suppository invention group on the rates of degrading between the two groups.2.In vitro studyThe growth of H8 cells with drug concentrations of 2,3,4,5 g·L-1 significantly inhibited in concentration-dependent and time-dependent manners,compared with blank group.According to Hoechst 33342 staining,nuclear chromatin condensations were observed with drug concentrations of 1,2,3,4,5 g·L-1 for 24h,and apoptotic bodies turned more when cells were treated with 3,4,5 g·L-1 drug concentrations increasingly.The early apoptotic rates of 1,2,3 g·L-1 drug concentrations were 23.73 ± 5.72,63.9 ± 3.59,71.53±1.59,detected by flow cytometry,gradually increased and had a statistically significant difference compared with the control group(6.53±0.85)(P<0.01).Caspase-3 and PARP proteins were consistently down regulated in H8 cells with Youdujing intervented and had a statistically significant difference compared with the control group.Conclusion1.For high risk HPV infection patients,Youdujing vaginal suppositories have the advantages of cleaning the infection.2.Youdujing can significantly inhibit the proliferation and induce early apoptosis in H8 cells,possibly by the way of caspase 3/PARP activation... |
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