| Objective:1.By observing the clinical efficacy of Yinyan Recipe in the treatment of high-risk Human papilloma virus(HPV)patients with primary cervical infection,evaluating its value in the treatment of high-risk cervical HPV infection,and providing evidence for clinical treatment.2.By observing the inhibitory effect of Yinyan Recipe on the expression of HPV16-E6 protein,and its effect on the expression of PI3K,p-PI3K,AKT and p-AKT,the inhibition and killing effect of Yinyan Recipe on HPV and its possible mechanism were discussed.Methods:1.clinical trialA total of 90 patients who met the inclusion criteria were selected from November 2021 to December 2022 in the Department of Gynecology,Department of Traditional Chinese Medicine and Department of Gynecology of Traditional Chinese Medicine in Cangzhou Hospital of Integrated TCM in Hebei.According to the random number table method,they were divided into the Yinyan Recipe group(n=30),the blank control group(n=30)and the Baofukang suppository group(n=30).The Yinyan Recipe group was given Yinyan Recipe capsule,and the Baofukang suppository group was given Baofukang suppository,both of them were given one capsule or suppository per vagina every other day for ten times a month,avoiding menstrual period,and the treatment was lasting for three months.The blank control group was only followed up for three months,without drug treatment.The TCM syndrome score of three groups was evaluated in the 4th month,and HPV and TCT test were rechecked in the 7th month,as well as colposcopy if necessary.2.Animal experimentsSix female C57BL/6 mice were randomly selected as the blank group,and the rest were used to establish the vaginal infection model of HPV16 E6over expression adenovirus mice.After 14 days of modeling,the mice were randomly divided into model group,Baofukang group,low,medium and high dose of TCM Yinyan Recipe groups,with 6 mice in each group.The mice in Baofukang group and low,medium and high dose of TCM Yinyan Recipe groups were respectively treated with 0.26g/kg of Baofukang suppository,0.075g/kg,0.15g/kg and 0.3g/kg of TCM Yinyan Recipe via vagina once a day.The mice in the model group and the blank group were given the same dose of normal saline vaginally once a day and the treatment was for 14 days.Immunofuorescence assays was used to observe the mean fluorescence intensity of EGFP,p-PI3K and p-AKT proteins in vaginal tissues,and Western blot was used to detect the expression of PI3K,p-Pi3k,AKT and p-AKT proteins in vaginal tissues.Results:1.Clinical study results(1)Compared with HPV negative conversion rate,the negative conversion rate of Yinyan Recipe group was 86.67%,Baofukang suppository group was 56.67%,and blank group was 40%.Yinyan Recipe group was significantly better than blank group and Baofukang suppository group,and the difference was statistically significant(P<0.05).There was no significant difference in effective rate between Baofukang suppository group and blank group(P>0.05).(2)The total scores of TCM syndromes in the three groups were compared after treatment,the improvement of TCM syndrome score in Yinyan Recipe group and Baofukang suppository were better than that in blank group,and Yinyan Recipe group was better than Baofukang suppository group,the differences were statistically significant(P<0.05);The total effective rate of Yinyan Recipe group was 93.3%,the total effective rate of Baofukang suppository group was 73.30%,and the total effective rate of blank group was 13.30%.The curative effect of Yinyan Recipe group and Baofukang suppository group were better than that of blank group,and the differences were statistically significant(P<0.05),the effect of Yinyan Recipe group was better than that of Baofukang suppository group(P<0.05).2.Results of experimental study(1)The titer of HPV16 E6 overexpression adenovirus was 5.25×10~5PFU/ml,and the infected C57BL/6 mice model was successfully established.(2)The mean fluorescence intensity of EGFP detected by Immuno fuorescence assays:compared with the blank group,the mean fluoresce nce intensity of EGFP in the vaginal tissue of the female mice in the model group was significantly increased(P<0.0001).Compared with t he model group,the average fluorescence intensity of EGFP in vaginal tissue of the mice in each dose of Yin Yan Recipe group and Baofuka ng group decreased(P<0.05).Compared with the high dose of Yinyan Recipe,the average fluorescence intensity of EGFP in vaginal tissue of the mice in the low and medium dose of Yinyan Recipe groups,wi th statistical significance(P<0.05).There was no significant difference in the average fluorescence intensity of EGFP between the low-dose group and medium-dose of Yinyan Recipe(P>0.05).There was no sign ificant difference in the average fluorescence intensity of EGFP betwee n Baofukang group and Yin Yan Recipe high-dose group(P>0.05).(3)The expressions of PI3K,p-PI3K,AKT and p-AKT proteins were detected by immunofluorescence and Western blot:Compared with blank group,the average fluorescence intensity and protein expression of PI3K,p-PI3K,AKT and p-AKT in model group were significantly up-regulated(P<0.0001).Compared with model group,the average fluorescence intensity and protein expression of PI3K,p-PI3K,AKT and p-AKT in vaginal tissues of mice in low-dose and medium-dose groups of Yinyan Recipe and Bafukang group were decreased,with statistical significance(P<0.01).Compared with Bafukang group,the average fluorescence intensity and protein expression of PI3K,p-PI3K,AKT and p-AKT in vaginal tissues of mice in low and medium dose groups of Yinyan Recipe were decreased,and the differences were statistically significant(P<0.05).There were no significant differences in the average fluorescence intensity and protein expression of PI3K,P-PI3K,AKT and p-AKT in vaginal tissues of mice in low and medium dose groups(P>0.05).There were no significant differences in the average fluorescence intensity and protein expression of PI3K,P-PI3k,AKT and P-AKT between Yinyan Recipe high-dose group and model groups(P>0.05).Conclusions:1.TCM Yinyan Recipe can effectively improve the negative conversion rate of high-risk HPV infection in female cervix,and significantly improve the TCM syndrome score.The clinical efficacy is better than Baofukang suppository,which is worthy of clinical promotion and application.2.In addition to inhibiting and killing HPV,TCM Yinyan Recipe could also block the tendency of CIN to develop into early cervical cancer.3.TCM Yinyan Recipe can effectively reduce the pathogenicity of HPV,and low and medium doses are effective,and there is no significant difference in their efficacy.The mechanism of action may be related to the inhibition of virus activation of PI3K/AKT pathway. |
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