Background & Aims: Endometriosis is a common female disorder that affects 6 to 10% of women. Histologically, endometriosis is the excessive appearance of fibrotic tissue around the stroma and endometrial glands. Fibrosis in endometriosis is regulated by the Wnt signaling pathway, which plays critical roles in various diseases (1). B-catenin plays the main role of wnt intracellular transformation pathway in vertebrates. B-catenin is a multifunctional protein that functions both as a transcriptional and as a membrane and cytoskeleton interface protein (2). Several studies have shown that aberrant activation of wnt/b-catenin signaling may play a role in the pathology of endometriosis (3, 4). On the other hand, it has been shown that the lack of movement and vitamins lead to the creation of various factors in the development of diseases, and various factors such as immune system defects, genetics, hormones and environmental conditions have an effect on the improvement of the disease. In order to reduce the symptoms of the disease, increase the healing process and improve the quality of life of the patients, researchers emphasize the implementation of appropriate interventions (5). It has been shown that aerobic exercises are one of the training methods to improve the performance of infertility and women's diseases. Studies show that low-intensity aerobic training, by creating a protective mechanism, reduces the expression of inflammatory cytokines, oxidative stress in uterine tissue, systemic inflammation, and as a result improves immune responses (14). It has also been shown that vitamin E reduces the pain caused by endometriosis by inhibiting the release of arachidonic acid and also inhibiting the conversion of arachidonic acid to prostaglandin through the action on phospholipase A2 and cyclooxygenar enzymes (18, 19). The results of the investigation by Giampolino et al. (2019) indicate that there is a direct relationship between the group of vitamins and the recovery of endometriosis (20). In general, endometriosis is a multifactorial disease with a complex pathophysiology, and most of the details of this disease are still unknown; Therefore, the need for extensive research to obtain information for the pathophysiology of this disease is essential. Therefore, according to the studies conducted, no research was found on the role of physical activity and vitamin consumption, especially vitamin E, on the Wnt/β-catenin pathway of endometriosis rats, so in the present study, the researcher seeks to answer this question. Does a period of regular swimming exercises and vitamin E consumption have an effect on the changes of Wnt/β-catenin pathway genes in ovarian tissue of endometriosis model mice?
Methods: Methods: This study was of an experimental type, and 30 8-week-old female Wistar rats with an average weight of 202/85±15/62 grams were purchased from Pasteur Institute. In order to create an endometriosis model, first, adult rats were anesthetized using ketamine and xylazine. Then, using a scalpel, an incision was made in the skin of the flank area in the pelvic area. After opening the abdominal muscle and the peritoneal area, first, the ovarian tissue was removed along with a part of the fallopian tube tissue. Then it was placed in a sterile container with one cc of PBS (phosphate buffer solution). Then each tissue was cut into one piece by one millimeter. The tissue pieces, which were 4 pieces for each mouse, were transplanted to the right pelvic muscle wall area, to the abdominal peritoneum area, to the anterior abdominal wall muscle area, and to the fat around the ovary. Then, the surgical area was sutured and the mice were transferred to the corresponding cage (22). After the induction of the model, the mice were randomly divided into 6 healthy control groups, sham, endometriosis, endometriosis + vitamin E, endometriosis + swimming, endometriosis + swimming + vitamin E. The endometriosis group remained two weeks after the creation of the model until the end of the study, and the healthy control group was maintained for 8 weeks, and the endometriosis + vitamin E group started receiving the supplement on a daily basis and in the form of gavage two weeks after the creation of the model. The amount of 200 kg/mg body weight of rats was maintained for 8 weeks until the end of the study. The rats of the endometriosis + swimming groups swam 5 days a week until the end of the research period in a water tank with dimensions of 50 x 50 x 100 cm with a temperature of 30-32 degrees Celsius during 8 weeks. . The duration of training in water was 30 minutes daily until the end of the training period (23). In order to eliminate the acute effect of swimming training and vitamin E consumption, animal sampling was done 48 hours after the last swimming training program and vitamin E consumption. For this purpose, first the animals were anesthetized and then sacrificed, and after sacrifice, the transplanted tissues related to the endometrial area were evaluated for histology and genetic studies. Real time PCR technique was used to examine the expression of wnt and β-catenin genes in each group. After the laboratory analysis of the tissue samples, considering the normality of the distribution of the data from the parametric test including the one-way analysis of variance test and Tukey's post hoc test at the significance level of P≤0.05 to investigate the changes in wnt gene expression and β-catenin was used. All statistical operations were performed using SPSS23 software.
Results: The mean and standard deviation of the research variables are presented in Table 2. Data analysis showed that there is a statistically significant difference between different research groups in the expression levels of Wnt and β-catenin genes (p═0.044, 0.048). The results of the follow-up test also showed that the expression levels of Wnt and β-catenin genes in the endometriosis group were significantly higher than in the healthy control group (p═0.035, 0.014). Swimming, vitamin E, and a combination of swimming + vitamin E were associated with a decrease in Wnt and β-catenin gene expression, and the endometriosis + swimming + vitamin E group had a significant decrease in β-catenin gene expression compared to endometriosis and endometriosis + vitamin E (p ═0.048, p ═0.05) and a non-significant decrease in Wnt gene expression (p═0.464, p═0.730).
Conclusion: The Wnt/β-catenin signaling pathway is an evolutionarily conserved cellular signaling cascade that plays a prominent role in various biological processes and the pathogenesis of many diseases, such as early embryonic development as well as organogenesis and adult tissue homeostasis. . Although the Wnt/β-catenin pathway exhibits many benefits, its dysregulation causes a wide range of diseases such as cancer, cardiovascular disease, and kidney disease (24, 25). Aberrant activation of Wnt/β-catenin has also been reported in patients with endometriosis or endometrial cancer. Abnormal Wnt/β-catenin signaling in the endometrium can lead to both embryo implantation failure and severe endometrial pathogenic changes such as endometrial cancer and endometriosis (26). Also, evidence shows that patients with endometriosis have higher levels of oxidative stress and inflammatory and pro-inflammatory cytokines (27). In order to reduce these symptoms, increase the healing process and the quality of life of patients, researchers have emphasized the implementation of appropriate interventions (28). Studies have shown that the consumption of food supplements and vitamins such as vitamin E, as well as exercising, controls the amount of estrogen in the body and helps to reduce the symptoms of endometriosis (29). In the current research, the effect of a swimming training program along with vitamin E consumption on the changes of Wnt/β-catenin pathway genes in endometriosis model mice was investigated. One of the important results of the current research is the abnormal increase in the levels of Wnt and β-catenin in the endometriosis model rats compared to the healthy group, which decreased with the intervention of exercise and vitamin E compared to the endometriosis group. In general, the results of the current research indicate that the regulation of Wnt and β-catenin gene expression in the normal state in the process of endometriosis can improve the level of this disease, and swimming exercises and the simultaneous consumption of vitamin E in the normal state of the Wnt /β-catenin pathway will be effective.