| Objective: 1.To analyze the clinical characteristics and risk factors of PGAD patients,and to observe the pathological characteristics and mechanical barrier changes of PGAD patients’ vaginal tissue;2.To establish a PGAD mouse model by simulating childbirth trauma and menopause,and to study functional and vaginal histomorphology.Evaluate the characteristics of PGAD model mice;3.Explore the role of estrogen in improving PGAD by regulating tight junction proteins.Methods Clinical trial: Patients who were scheduled to undergo hysterectomy and vaginal surgery in the Department of Gynecology of The Hospital of Ningxia Medical University from January 2021 to October 2022 were selected,and all subjects were subject to a one-to-one questionnaire survey by members of the research group after obtaining their informed consent;according to FSFI diagnosis Scale and PGAD diagnostic criteria,all participants were divided into PGAD group and non-PGAD group.Normal vaginal tissues from vaginal stumps were collected from all subjects after surgery for morphological staining and immunohistochemical staining and observation analyze.Animal experiments:8-week-old female C57BL6 mice were used and divided into sham operation group(sham),model group(PGAD)and estrogen treatment group(E2).In the sham group,a median incision above the pubic symphysis was made,and the abdominal cavity was opened and then closed;the mice in the PGAD group were established by bilateral ovariectomy combined with balloon vaginal expansion;the mice in the E2 group were given intravaginal estrogen cream on the basis of the PGAD model.After administration and treatment for 28 days,the mice in the PGAD group were administered vaginally with the same dose of Vaseline gel.On the 29 th day after modeling,the changes in sexual behavior and sexual response related indicators of mice in different groups were observed through behavioral studies,and then the blood and vaginal tissues were collected to detect the levels of serum sex hormones and morphology in different groups.The pathological changes of vaginal were observed by staining,and the expressions of tight junction protein in vaginal tissue were detected by immunohistochemistry and Western blotting.Results1.A total of 92 effective subjects were included in this study,including 59 PGAD patients and 33 cases of normal sexual function.The overall incidence of PGAD was 64.13%.The average FSFI score of the two groups of subjects was(28.63±2.57 vs.17.69±4.58,P<0.05);in libido(4.32±0.44 vs.2.63±0.20,P=0.032);sexual arousal(3.97±0.36 vs.2.20±0.41,P=0.046);Lubrication(4.52±0.50 vs.2.23±0.21,P = 0.021);sexual satisfaction(4.45±0.56 vs.2.62±0.90,P = 0.029)were statistically significant(P<0.05),while there were no significant differences(P>0.05)in orgasm(4.78±0.41 vs.4.35±0.47,P=0.36)and dyspareunia(3.64±0.81 vs.3.26±0.83,P=0.16).Through Logistic regression analysis,it was found that BMI(overweight),menopause,number of births(>1),underlying diseases,and vaginal diseases(cervical tumors)were important factors affecting sexual dysfunction(P<0.05);2.Vagina atrophy occurred in PGAD patients,the thickness of epithelial and muscular layer was thinner than sham group(P<0.05),the muscle fibers were broken,and the number of microvessels decreased compared with sham group(P<0.05);3.In terms of vaginal mechanical barrier function,there was no significant difference in the expression of MMP-9 in the vaginal tissues of the two groups of subjects(P>0.05).In the vaginal tissues of PGAD patients,ZO-1 and Occludin were mainly expressed in the vaginal epithelial and muscular layers,the content was lower than that in the normal group(P<0.05),Cladin-1 was mainly expressed in the epithelial layer,and the expression in the PGAD group was also lower than that in the non-PGAD group(P<0.05);4.In the PGAD mouse model,the vaginal weight of the mice decreased(P<0.05),and the levels of E2,T and DA in the serum decreased(P<0.05).After estrogen treatment,the vaginal weight and sex hormone levels increased compared with the PGAD group(P<0.05);In terms of sexual behavior,the number of sexual behavior rejections of male mice in PGAD group increased(P<0.05),the number of behaviors before mating decreased(P<0.05),and there was no significant difference in the return time after mating(P<0.05),after estrogen treatment,the number of rejections of sexual behavior decreased(P<0.05),and the behavior before mating increased(P<0.05);in terms of sexual response,the degree of vaginal lubrication of mice in the PGAD group decreased,and after estrogen treatment The degree of lubrication increased,but still less than that of the sham group(P<0.05).In terms of vaginal tension,the contraction of the vagina in the PGAD group became irregular,and the contraction peak decreased.After estrogen treatment,the vaginal contraction became more regular than that in the PGAD group;5.After modeling,the epithelial layer and muscular layer of the vagina in the PGAD group became thinner,the degree of fibrosis was deeper than that of the sham group,the glycogen content was lower than that of the sham group,and the number of microvessels in the vaginal tissue was also lower than that of the sham group(P<0.05);After 1 month of estrogen treatment,the vagina of the mice was thicker than PGAD group,the degree of fibrosis was alleviated,the content of glycogen increased,and the number of microvessels recovered compared with that of the PGAD group(P<0.05);6.In terms of the vaginal mechanical barrier,the contents of three tight junction proteins ZO-1,Occludi and Claudin-1 in the vaginal tissue of the mice in the PGAD group decreased(P<0.05).Further quantitative analysis of the above three proteins by Western blotting also yielded consistent results,that is,compared with the sham group,tight junctions in vaginal tissues of PGAD mice The protein content is reduced,and estrogen treatment can increase the expression of tight junction proteins.Conclusion1.The clinical characteristics of PGAD patients are mainly low libido,sexual arousal disorder,difficulty in vaginal lubrication and decreased sexual satisfaction.Body mass index(overweight),menopause,number of births(>1),underlying diseases and vaginal diseases are PGAD;2.The pathological characteristics of vaginal tissue in patients with PGAD are mainly manifested as the general atrophy of vaginal epithelial layer and muscular layer thickness,decreased number of microvessels,deepened fibrosis,decreased expression of three tight junction proteins,and impaired vaginal mechanical barrier function;3.The PGAD mouse model established by simulating childbirth trauma and menopause can accurately reflect the clinical and pathological characteristics of PGAD.The sexual response ability of PGAD model mice is significantly reduced,vaginal tissue atrophy,vaginal contraction performance is reduced,and the degree of fibrosis Deepening,the mechanical barrier function of the vagina is destroyed,so a scientific and reproducible animal model can be established through vaginal dilation and ovariectomy;4.Estrogen can improve the symptoms of PGAD,regulate the expression of tight junction proteins,and repair the mechanical barrier of vaginal tissue,which may be an important reason for the clinical treatment of PGAD with estrogen. |
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