Induction Of Endometriosis Research Articles

Possible therapeutic effect of royal jelly on endometriotic lesion size, pain sensitivity, and neurotrophic factors in a rat model of endometriosis.

Endometriosis is the abnormal growth of endometrial tissue. The goals of the study are: (1) Is any correlation between endometriosis pain and neurotrophins in the serum, dorsal root ganglion (DRG), and peritoneal fluid (PF) in rat models of experimental endometriosis?, (2) Possible therapeutic effects of royal jelly (RJ) on pain scores, size of endometriotic lesion, and neurotrophic factors. Forty‐eight Sprague Dawley female rats weighing 205.023 ± 21.54 g were maintained in a standard condition. The rats were randomly divided into one of the six groups: Control (no intervention), Sham‐1 (remove of uterine horn), RJ (administration of 200 mg/kg/day RJ for 21 days), Endometriosis (induction of endometriosis), Treatment (induction of endometriosis+administration of 200 mg/kg/day RJ for 21 days), and Sham‐2 (induction of endometriosis+administration of water). Formalin test performed for pain evaluation. The levels of Brain‐derived neurotrophic factor (BDNF), nerve growth factor (NGF), substance P, and calcitonin gene‐related peptide (CGRP) were measured by enzyme‐linked immunosorbent assay. The mean pain scores in all three phases of the formalin test were significantly increased by endometriosis induction (p < 0.05). The concentrations of BDNF, NGF, and CGRP in DRG of the endometriosis group were significantly higher than these factors in the Control, Sham‐1, and RJ groups (p < 0.05). RJ could significantly (p < 0.001) decrease the mean lesion size and the mean pain score in the late phase (p < 0.05). The present results determine that endometriosis pain may be related to nervous system neurotrophic factors. Treatment with RJ could decrease the size of endometriosis lesions as well as pain scores. The findings may shed light on other complementary and alternative remedies for endometriosis.

New rat to mouse xenograft transplantation of endometrium as a model of human endometriosis.

BackgroundEndometriosis can lead to infertility. Since there is no definitive treatment for endometriosis, animal modelling seems necessary to examine the possible treatments. Mouse endometrium cannot be separated for endometriosis induction. In addition, transplantation of uterus into the abdominal viscera to induce endometriosis causes organ damage. In this study, we defined a new model of endometriosis leading to separability of endometrium and a safe anatomical region for transplantation.MethodsForty female mice were allocated to 5 groups: 1, sham; 2, allograft uterus transplantation of mice to anterior abdominal wall of mice; 3, allograft uterus transplantation of mice to mesentery of mice; 4, xenograft endometrial transplantation of rat to anterior abdominal wall of mice; 5, xenograft endometrial transplantation of rat to mesentery of mice. Adult female rats with a previous pregnancy experience were selected and placed in the vicinity of male rats for 2 weeks to induce estrogen secretion and increase endometrial thickness.ResultsIn the 4th group of animals, compared to sham, the peritoneal concentrations of VEGF‐A, TNF‐α, NO, MDA, and serum levels of CA‐125 and IL‐37 were increased and total body weight was decreased, while weight and size of endometrial lesions were increased significantly (P < .05). Genes expression of HOXA10 and HOXA11 were decreased significantly (P < .05) in groups 2 and 4 compared to sham.ConclusionsXenograft transplantation of endometrium from rat to anterior abdominal wall of mice can potentially mimic human endometriosis morphologically, histologically, and genetically.

THE EFFECTS OF ETANERCEPT AND CABERGOLINE ON ENDOMETRIOTIC IMPLANTS, UTERUS AND OVARIES IN RAT ENDOMETRIOSIS MODEL

The pathophysiology of endometriosis is still unknown and treatment options remain controversial. Searches focus on angiogenesis, stem cells, immunologic and inflammatory factors. This study investigated the effects of etanercept and cabergoline on ovaries, ectopic, and eutopic endometrium in an endometriosis rat model. This randomized, placebo-controlled, blinded study included 50 rats, Co(control), Sh(Sham), Cb(cabergoline), E(etanercept), and E + Cb(etanercept + cabergoline) groups. After surgical induction of endometriosis, 2nd operation was performed for endometriotic volume and AMH level. After 15 days of treatment: AMH level, flow cytometry, implant volume, histologic scores, immunohistochemical staining of ectopic, eutopic endometrium, and ovary were evaluated at 3rd operation. All groups had significantly reduced volume, TNF-α, VEGF, and CD 146/PDGF-Rβ staining of endometriotic implants comparing to the Sh group (p < 0.05).TNF-α staining of eutopic endometrium in all treatment groups was similar to Sh and Co groups (p > 0.05). E and E + Cb groups significantly decreased TNF-α staining in the ovary comparing to Sh, Co, and Cb groups (p < 0.05). All treatment groups had significantly higher AFC compared to the Sh group. CD25+ Cells’ median percentage was significantly increased in the E + Cb group compared to Co, Sh, Cb, and E group. E + Cb group had a significantly higher CD5+ Cells’ level than the Co group (p = 0.035). In conclusion; Etanercept and/or Cabergoline decreased volume, TNF-α, VEGF, and CD 146/PDGF-Rβ staining of the ectopic endometrial implant. E and E + Cb treatment decreased TNF-α levels in the ovary. E + Cb also increased peripheral blood CD25+ & CD5+ Cell’s.

Astragaloside IV exerts anti-inflammatory role in endometriosis by downregulating TLR4/NF-κB pathway

Purpose: To investigate the effect of astragaloside IV administration on the inflammatory response in endometriosis and the underlying mechanism of action.&#x0D; Methods: Mice were divided into two groups: endometriosis (EMs) mice and control mice (n = 12). EMs induction in mice was achieved by transplantation of mouse uterine tissue. The same procedure was performed in control mice except that a separate suture was inserted instead of endometrial tissue. After 5 weeks, EMs mice were treated with or without astragaloside IV (AIV). The tissue lesions in EMs and control mice were stained with hematoxylin and eosin staining. The activation of toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) p65 signaling was evaluated by western blot, while expression of inflammatory cytokines was evaluated by quantitative real-time-polymerase chain reaction (qRT-PCR), and enzyme-linked immunosorbent assay (ELISA).&#x0D; Results: Astragaloside IV repressed the inflammation of murine Ems lesions, and also dampened the activation of TLR4/NF-κB signaling in vivo and vitro (p &lt; 0.01 and p &lt; 0.001, respectively). In addition, the expression levels of inflammatory cytokines (IL-1β, IL-6, Ccl-2, and TNF-α) decreased following AIV treatment in vivo.&#x0D; Conclusion: The results indicate that TLR4/NF-κB signaling pathways are closely related to the inhibition of Ems inflammation by astragaloside IV. Thus, astragaloside IV may be a novel drug for the prevention and treatment of endometrioses.

Loss of Cxcr4 in Endometriosis Reduces Proliferation and Lesion Number while Increasing Intraepithelial Lymphocyte Infiltration

Hyperactivation of the CXCL12-CXCR4 axis occurs in endometriosis; the therapeutic potential of treatments aimed at global inhibition of the axis was recently reported. Because CXCR4 is predominantly expressed on epithelial cells in the uterus, this study explored the effects of targeted disruption of CXCR4 in endometriosis lesions. Uteri derived from adult female mice homozygous for a floxed allele of CXCR4 and co-expressing Cre recombinase under control of progesterone receptor promoter were sutured onto the peritoneum of cycling host mice expressing the green fluorescent protein. Four weeks after endometriosis induction, significantly lower number of lesions developed in Cxcr4-conditional knockout lesions relative to those in controls (37.5% vs. 68.8%, respectively). In lesions that developed in Cxcr4-knockout, reduced epithelial proliferation was associated with a lower ratio of epithelial to total lesion area compared with controls. Furthermore, while CD3+ lymphocytes were largely excluded from the epithelial compartment in control lesions, in Cxcr4-knockout lesions, CD3+ lymphocytes infiltrated the Cxcr4-deficient epithelium in the diestrus and proestrus stages. Current data demonstrate that local CXCR4 expression is necessary for proliferation of the epithelial compartment of endometriosis lesions, that its downregulation compromises lesion numbers, and suggest a role for epithelial CXCR4 in lesion immune evasion.

Evidence for Beneficial Effects of Voluntary Exercise in an Endometriosis‐Stress Animal Model

Endometriosis is a chronic gynecological disorder characterized by peritoneal inflammation, infertility, chronic pelvic pain, fibrosis, and growth of endometrial tissue outside the endometrial cavity. In previous studies we found that exposure to stress exacerbates the development and severity of endometriosis. Exercise has been shown to confer beneficial effects in many chronic conditions. We previously demonstrated that exposure to voluntary exercise prior and after the induction of endometriosis in an animal model can improve the condition. However, it is unclear whether exercise can also alleviate the stress-induced exacerbation of endometriosis. Hypothesis Voluntary exercise interventions can promote and help to ‘realign/reset’ dysregulation of the brain-gut axis caused by psychological stress resulting in reduced symptoms in animals with endometriosis. Methods Endometriosis (Endo) was induced in female Sprague Dawley Rats by implanting four pieces of uterine tissue next to the intestinal mesentery on day 0, while Sham controls received sutures only. Abdominal circumference and BMI were measured and calculated respectively for all animals before surgery. All animals were exposed to water avoidance stress at day 14 after surgery for seven consecutive days (1hr/day). Endo (n=10) and Sham (n=10) controls received no exercise. Endometriosis-exercise (EEX; n=10) and sham-exercise (SHEX; n=10) had access to a running wheel after surgery until time of sacrifice (day 60). At sacrifice, the abdominal circumference and BMI was measured and calculated, and the developed vesicles and colon were collected and stored for future analysis. Results Exercised groups consumed more food when compared to their respective controls (Endo vs EEX p<0.001; Sham vs SHEX p<0.01) but tended to gain less weight overall. No difference was observed in the distance or speed travelled between EEX and SHEX. Endo had significantly higher percent of fat/body weight compared to sham groups (p<0.001), and this was reversed by exercise. Endo animals had significantly increased abdominal circumference (p<0.05) and BMI (p<0.05) at sacrifice compared to the day of surgery; these differences were not significant in the other groups. Endo animals developed vesicles in 75.0% of the implants with a total length of 11.08±1.74mm/animal and total vesicle weight of 0.12±0.02g/animal. No vesicles developed in Sham or SHEX. Exposure to exercise significantly decreased the number of vesicles that developed (42.5% p<0.05), as well as vesicle size (5.86±1.57mm, p<0.05), and weight (0.05±0.02g, p<0.05) per animal. Endo had increased colonic macroscopic damage compared to Sham (p<0.001), and exercise groups (p<0.001). Conclusions Our results suggest that voluntary physical activity might alleviate the aggravated endometriosis associated symptomatology which occurs during stress conditions. Overall, this study supports the potential for further investigation of voluntary exercise as a complementary therapy in patients with endometriosis.

Preoperative and perioperative intervention reduces the risk of recurrence of endometriosis in mice caused by either incomplete excision or spillage and dissemination

Research questionCan preoperative or perioperative intervention reduce the risk of recurrence of endometriosis caused by either incomplete excision or spillage and dissemination? DesignA mouse model of endometriosis recurrence caused by spillage and dissemination was first established using 24 female Balb/c mice. The spillage and dissemination model was used to test the efficacy of preoperative use of ketorolac, perioperative use of aprepitant and combined use of propranolol and andrographolide in a prospective, randomized mouse experiment involving 75 mice. The efficacy of these preoperative and perioperative interventions in a mouse recurrence model caused by incomplete excision was also tested using 72 mice. In all experiments, the baseline body weight and hotplate latency of all mice were measured and recorded before the induction of endometriosis, before the primary surgery and before sacrifice. In addition, all lesions were excised, weighed and processed for quantification and immunohistochemistry analysis of E-cadherin, α-SMA, VEGF, ADRB2 and putative markers of recurrence PR-B, p-p65, as well as Masson trichrome staining. ResultsAll interventions substantially and significantly suppressed the outgrowth of endometriotic lesions and reduced the risk of recurrence caused by either spillage and dissemination or incomplete excision (P = 0.0007 to 0.042). These interventions also significantly attenuated the generalized hyperalgesia, inhibited the staining of α-SMA, p-p65, VEGF and ADRB2 but increased staining of E-cadherin and PR-B, resulting in reduced fibrosis. ConclusionGiven the excellent safety profiles of these drugs, these data strongly suggest that preoperative and perioperative intervention may potentially reduce the risk of endometriosis recurrence effectively.

In vivo effects of AZD4547, a novel fibroblast growth factor receptor inhibitor, in a mouse model of endometriosis.

Endometriosis is a chronic disease, characterized by the growth of endometrial‐like cells outside the uterine cavity. Due to its complex pathophysiology, a totally resolving cure is yet to be found. The aim of this study was to compare the therapeutic efficacy of AZD4547, a novel fibroblast growth factor receptor inhibitor (FGFRI), with a well‐characterized progestin, etonogestrel (ENG) using a validated in vivo mouse model of endometriosis. Endometriosis was induced by transplanting uterine fragments from donor mice in proestrus into the peritoneal cavity of recipient mice, which then developed into cyst‐like lesions. AZD4547 and ENG were administered systemically either from the day of endometriosis induction or 2‐weeks post‐surgery. After 20 days of treatment, the lesions were harvested; their size and weight were measured and analyzed histologically or by qRT‐PCR. Stage of estrous cycle was monitored throughout. Compared to vehicle, AZD4547 (25 mg/kg) was most effective in counteracting lesion growth when treating from day of surgery and 2 weeks after; ENG (0.8 mg/kg) was similarly effective in reducing lesion growth but only when administered from day of surgery. Each downregulated FGFR gene expression (p < 0.05). AZD4547 at all doses and ENG (0.008 mg/kg) caused no disturbance to the estrous cycle. ENG at 0.08 and 0.8 mg/kg was associated with partial or complete estrous cycle disruption and hyperemia of the uteri. AZD4547 and ENG both attenuated endometriotic lesion size, but only AZD4547 did not disrupt the estrous cycle, suggesting that targeting of FGFR is worthy of further investigation as a novel treatment for endometriosis.

Effects of atorvastatin and resveratrol against the experimental endometriosis; evidence for glucose and monocarboxylate transporters, neoangiogenesis

The current study was conducted to investigate the therapeutic effects of atorvastatin (ATV) and resveratrol (RVT) in sole and simultaneous forms of administration against the symbiosis between glucose transporters 1 and 3 (GLUT-1 and GLUT-3), monocarboxylate transporters 1 a and 4 (MCT-1 and MCT-4) and neovascularization in ectopic endometrial tissue (EET). For this purpose, the experimental endometriosis was induced in 24 virgin female Wistar rats, and then the rats were divided into non-treated endometriosis-induced (ENDO-sole), AVT-treated (5 mg kg-1), RVT-treated (40 mg kg-1) and AVT +RVT-treated groups (n = 6 rats in each group). Following 28 days from the experimental endometriosis induction, the EETs were collected and the EETs size, neovascularization ratio, and expression levels of GLUT-1, GLUT-3, MCT-1, and MCT-4 were analyzed by qRT-PCR and immunohistochemistry (IHC). The AVT and RVT sole and simultaneous-treated animals exhibited decreased EET sizes and neovascularization. Moreover, the mRNA levels of GLUT-1, GLUT-3, MCT-1, and MCT-4, as well as GLUT-1+, GLUT-3+, and MCT-4+ cells distribution per mm2 of tissue were decreased in AVT and RVT sole and simultaneous-treated groups. Our findings showed that the AVT and RVT, especially in the simultaneous form of administration, could decrease the neovascularization development in the EETs by suppressing the GLUTs (1 and 3) and MCTs (1 and 4) expressions. Therefore, it can be concluded that the simultaneous administration of AVT and RVT can inhibit the EET's establishment and development through suppressing glycolysis and neovascularization.

Murine Models of Endometriosis: A Systematic Review

Objectives: This review inspects the usage of animal models and the practical solutions of this method for different challenges of endometriosis. The objectives of the study are to determine and compare the histopathology, biomarkers, and development of endometrial lesions in murine homologous and heterologous endometriosis models. Methods: The literature search was performed on PubMed, Scopus, Web Cochrane, and EMBASE from January 1990 to January 2019. Experimental articles in which the establishment of the endometriosis model had been proven through the examination of size, weight, number of implants, adhesion, histologic score, and altered biomarker were eligible for inclusion. Results: Based on type of induction, articles were categorized into two groups: heterologous-induced method (n=5) and autologousinduced method (n=13). In general, in case of establishing the heterologous induction method is less reliable than the autologous induction method. Conclusions: Using human endometrial tissues for endometrial inductions is possible in heterologous models under immunosuppression, which is more suitable for therapeutic studies, but time limitation considerations are mandatory for this type of model. Homologous endometriosis inductions cause larger endometrial lesions, biomarkers, and reproduction rate changes similar to those occurring in humans. Similarities make this method more appropriate for pathogenesis and genetic studies and also observe the impact of endometriosis on the next generation. Choosing an appropriate model for the induction of endometriosis is dependent on the purpose of each study.

Assessing Pain Behavioral Responses and Neurotrophic Factors in the Dorsal Root Ganglion, Serum and Peritoneal Fluid in Rat Models of Endometriosis.

Objective: Pain is the most frequently reported symptom involving in endometriosis. The alterations of neurotrophic factors and certain neuropeptides in the dorsal root ganglion (DRG), as well as serum and peritoneal fluid (PF), were evaluated in rat models of endometriosis. Materials and methods: Twenty-four Sprague Dawley female rats were selected and maintained in a standard condition with 12 hours’ dark-light cycles. All the rats were randomly assigned to 3 groups: Control (intact rats); Sham (the operation was conducted without endometriosis induction); and Endometriosis (endometriosis induction was performed). The formalin test was performed for all groups on the first and the 21st day of the study. The assessments of Brain-Derived Neurotrophic Factor (BDNF), Nerve Growth Factor (NGF), Calcitonin Gene-Related Peptide (CGRP), and Substance P levels were carried out by enzyme-linked immunosorbent assay (Elisa). The data were analyzed by One-Way ANOVA. The Tukey’s test was used as post-hoc. Results: Endometriosis induction significantly increased the mean pain scores in the endometriosis group in all three phases of the formalin test. The concentrations of DRG-CGRP (p=0.035), BDNF (p<0.001), and NGF (p=0.006) in the endometriosis group were significantly higher than that of the other groups while serum-BDNF (p<0.001), Substance P (p=0.009), and NGF (p=0.015) were significantly lower in endometriosis group compared to other groups. The concentrations of PF-BDNF (p=0.025) and Substance P (p=0.009) were significantly lower than those of other groups. Conclusion: The present results delineate that endometriosis induction could lead to hyperalgesia. This may be related to the significant increases in the BDNF, NGF, and CGRP in DRG.

A Syngeneic Murine Model of Endometriosis using Naturally Cycling Mice.

Endometriosis is a leading cause of pelvic pain and infertility. It is defined by the presence of endometrial tissue in extrauterine locations. The development of novel therapies and diagnostic tools for endometriosis has been limited due in part to challenges in studying the disease. Outside of primates, few mammals menstruate, and none develop spontaneous endometriosis. Rodent models are popular but require artificial induction of endometriosis, with many utilizing either immunocompromised mice or surgically induced disease. Recently, more attention has been given to models involving intraperitoneal injection. We present a murine model of endometriosis that integrates several features of existing endometriosis models into a novel, simplified system that relies on microscopic quantification in lieu of subjective grading. In this model, we perform hormonal stimulation of donor mice, intraperitoneal injection, systematic abdominal survey and tissue harvest, and histologic quantification that can be performed and verified at any time after necropsy. This model requires minimal resources and training; does not require expertise by lab technicians in murine survival surgery or in the identification of gross endometriotic lesions; can be used in immunocompromised, immunocompetent, and/or mutant mice; and reliably creates endometriotic lesions that are histologically consistent with human endometriotic disease.

The Role of Ankaferd Blood Stopper and Oxytocin as Potential Therapeutic Agents in Endometriosis: A Rat Model.

To evaluate the potential effect of Ankaferd Blood Stopper (ABS) and oxytocin (OT) in an experimental endometriosis model, 18 female Sprague Dawley rats were used in this study. The animals were divided randomly into three groups after surgical induction of endometriosis: group 1: control group (isotonic NaCl, 1 mL/kg/day, intramuscular, n=6); group 2: OT group (OT, 80 U/kg/day, intramuscular, n=6); group 3: ABS group (ABS, 1.5 mL/kg/day, intraperitoneal, n=6). Each group was treated for four weeks (two times per week). Volumes of endometriotic explants were measured in biopsy samples for histopathological analysis. Vascular endothelial growth factor (VEGF), monocyte chemotactic protein-1 (MCP-1), and tumour necrosis factor (TNF-α) levels were measured in plasma and peritoneal fluid. Endometriotic explant volumes were significantly decreased after OT administration (P<0.0001). The epithelial score was significantly decreased in both treatment groups compared to the control group (P<0.05). TUNEL immunohistochemistry showed more apoptotic changes in the endometriosis foci (gland epithelium and surrounding tissue) in the OT group than in the control group (P<0.05). The levels of VEGF, MCP-1, and TNF-α were significantly reduced in the OT group (P<0.05), whereas no significant changes in protein levels were found in the ABS-applied group. The results indicate that OT has greater potential as a therapeutic agent in experimentally induced peritoneal endometriosis, where ABS, which is a VEGF modulator, appears to act through different mechanisms to show its palliative effects on a rat model of peritoneal endometriosis.

Anti-endometriotic effect of &lt;i&gt;Angelica sinensis&lt;/i&gt; (Oliv.) Diels extract in human endometriotic cells and rats

Purpose: To study the anti-endometriotic effect of Angelica sinensis (Oliv.) Diels extract (ASDE) in human endometriotic cells and rats.Method: Forty female rats were randomly divided into four groups (10 rats/group): control, endometriosis+danazol, endometriosis+high dose of ASDE and low dose of ASDE. The rats were orally administered either vehicle (200 μL of PBS) alone or ASDE (140, 280 and 560 mg/kg/day) for 5 weeks. Danazol was used as the control drug. After induction of endometriosis for 4 weeks, the rats were sacrificed by cervical dislocation and the peritoneum and visceral organs examined visually to measure the number of endometriotic lesions. Serum levels of cancer antigen 125 (CA-125) and interleukin 13 (IL-13), interleukin 18 (IL-18) and tumor necrosis factor-alpha (TNF-α) of peritoneal fluids of rats were measured using ELISA kits. Western blot assay was performed to measure the levels of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) expressions after 24 h of treatment with ASDE (30, 60, and 120 μg/mL).Results: ASDE-treated rats displayed reduced numbers of total endometriotic lesions when compared with vehicle-treated controls (p &lt; 0.01). When the rats were treated with high dose of ASDE, serum CA-125 level, as well as IL-18 and TNF-α levels in peritoneal fluids were significantly lower than that of the control group (p &lt; 0.01); however, IL-13 level in peritoneal fluids was significantly higher than that of the control group (p &lt; 0.01). ASDE treatment significantly suppressed the levels of MMP-2 and MMP-9 protein in 11Z cell (p &lt; 0.01).Conclusion: The results reveal that ASDE exhibits significant anti-endometriotic effect by inhibiting inflammatory factors in rats. Thus, the plant extract can potentially be developed for the clinical management of endometriosis.&#x0D; Keywords: Angelica sinensis, Endometriosis, Cancer antigen, Endometriotic lesions, Matrix metalloproteinase

Enriched Environment Decelerates the Development of Endometriosis in Mouse.

We tested the hypothesis that enriched environment (EE), consisting of enlarged space, and increased physical activity and social interactions, hinders the development of endometriosis through attenuated adrenergic signaling, enhanced autophagy, and reduced leptin levels. Two mouse experiments were performed. In Experiment 1, 40 female Balb/C mice were randomly divided into four equal-sized groups, the SE (standard environment), EE, p-EE (EE instituted after endometriosis induction), and the d-EE (SE housing but received uterine fragments from EE donors) groups. Housing intervention was initiated 3weeks before the induction of endometriosis and continued for 3weeks after induction. In Experiment 2, 20 female mice were randomly divided into SE and EE groups, and the plasma leptin levels were measured. We measured lesion weight and hotplate latency and performed Masson trichrome staining as well as immunohistochemistry analysis of β2 adrenergic receptor (ADRB2), dopamine receptor D2 (DRD2), vascular endothelial growth factor (VEGF), and microtubule-associated protein light chain 3 (LC3). We found that EE reduced the lesion weight by 40.8% as compared with SE mice, but the reduction in p-EE and d-EE mice did not reach statistical significance. EE significantly reduced staining levels of ADRB2 and VEGF as well as the extent of lesional fibrosis but increased staining levels of LC3 and DRD2 in lesions as compared with the SE group. EE mice had reduced plasma leptin levels as compared with SE mice. Thus, EE decelerates the development of endometriosis and fibrogenesis and improved generalized hyperalgesia, possibly through increased DRD2 expression but decreased expression of ADRB2 and VEGF as well as enhanced autophagy and reduced leptin level.

Animals with Endometriosis Experience Beneficial Effects and Localized Modulation of Cytokine Levels when Exposed to Voluntary Wheel Running

A physically active lifestyle has been shown to confer preventive health benefits in many chronic conditions, in part by enhancing immunological health. Prolonged moderate exercise increases secretion of IL‐6 that may also promote secretion of the anti‐inflammatory cytokine IL‐10. Endometriosis is a chronic gynecological disorder characterized by peritoneal inflammation, chronic pelvic pain, fibrosis, and growth of endometrial tissue outside the endometrial cavity. We recently demonstrated protective effects of prior exercise in this condition, however it is still unclear whether physical activity can alleviate endometriosis that has already developed.HypothesisVoluntary physical exercise can promote immune strength helping to reduce symptoms in animals with endometriosis.MethodsEndometriosis was induced in female Sprague Dawley Rats by implanting four pieces of uterine tissue next to the intestinal mesentery on day 0, while Sham controls received sutures only. Endo (n=11) and Sham controls (Sham; n=11) received no exercise. Another group of endometriosis (EEX; n=10) and sham animals (SHEX; n=10) had access to a running wheel after induction of endometriosis until time of sacrifice at day 60. At sacrifice the developed vesicles, colon, serum and peritoneal fluid were collected and stored for future analysis.ResultsEndo animals developed vesicles in 79.55% of the implants with an average total area of 71.02±15.50mm2 and average developed vesicle weight of 0.12±0.04g. No vesicles developed in Sham or SHEX. Endo had significantly increased macroscopic colonic damage (p&lt;0.0001 vs Sham and SHEX). Exposure to exercise significantly decreased the number of vesicles that developed (35.00%, p&lt;0.01), as well as their size (10.23±4.24mm2, p&lt;0.01), and weight (0.03±0.01g, p&lt;0.05). EEX had significantly less macroscopic (p&lt;0.0001) and microscopic (p&lt;0.05) colonic damage compared to Endo. Exercise increased the release of various cytokines and chemokines including MIP1 (p&lt;0.01), IL‐10, IP10 (p&lt;0.05), RANTES, and IL‐6 (p&lt;0.05) in peritoneal fluid compared to Endo. Changes in cytokine levels at the systemic level were less pronounced.ConclusionsOur results suggest that voluntary physical activity might alleviate endometriosis associated progression of symptomatology, and enhance immune competency by impacting various inflammatory cytokines and angiogenic factors. Taken together, this opens the potential for further exploration of voluntary exercise as a complementary therapy in the patient population for whom current treatments are still non‐curative with many side effects.Support or Funding InformationSupported in part by R15AT009915, R25GM082406, U54MD007579.

Voluntary Exercise Impacts Body Composition, Leptin and Vesicle Size in a Rat Model of Endometriosis

Endometriosis is a chronic inflammatory condition caused by migration of endometrial tissue outside the uterine cavity resulting in dysmenorrhea, chronic pelvic pain and infertility. Current hormonal or surgical treatments for this debilitating condition are mostly focused towards infertility and pain relief. Physical activity has been shown to modulate body fat related factors, while having beneficial effects on chronic inflammatory conditions, and more recently on pain management.HypothesisExposure of animals with endometriosis to a voluntary exercise intervention will impact body composition and reduce vesicle development in an animal model of endometriosis.MethodsTissue from the uterine horn was sutured on the intestinal mesentery of female Sprague‐Dawley rats for endometriosis induction (day 0), while the Sham rats received sutures with no implants. Sham control rats (Sham; n=11) and an Endo group (Endo; n=11) were not exposed to exercise. Another Endo group (EEX; n=10) and Sham group (SHEX; n=10) had free access to running wheels after surgery until day of sacrifice (day 60). The Scurry Activity System software was used to collect the running wheel activity data (speed, distance and intensity). Weight change and food consumption were measured throughout the protocol. At sacrifice, vesicles were collected and measured, along with mesenteric fat, serum, and muscles.ResultsAll animals reached a minimum running distance of 2 km/day by day 7 after surgery. Regardless of endometriosis, there were no differences in average total distance ran per animal per group (318.48 ± 62.52 km in SHEX vs 471.22 ± 75.79 km in EEX) or average speed (2.23 ± 0.53 m/min in SHEX vs 3.08 ± 0.52 m/min EEX). Exposure to exercise significantly reduced the average weight of the vesicles (p&lt;0.05) and average total area (p&lt;0.01) when compared to the Endo no exercise group. Both groups exposed to running wheels consumed significantly more food after surgery when compared to the no exercise groups (p&lt;0.001). The exercise rats gained significantly less weight throughout the protocol, although absolute weight at sacrifice was not significantly different across all groups. Endo animals had significantly higher amounts of mesenteric fat when compared to Sham (p&lt;0.01), while exercise reversed this effect (p&lt;0.001). Exercise significantly decreased leptin serum concentrations (p&lt;0.001 vs Sham; p&lt;0.05 vs Endo). The muscle/fat ratio was also increased in the exercised rats when compared to controls in gastrocnemius (p&lt;0.05) and soleus (p&lt;0.01).ConclusionsAccess to voluntary physical activity after endometriosis induction promotes decreased body fat composition and ameliorates the development of vesicles. The surgical induction of endometriosis did not impact ability to run. The role of leptin in endometriosis is still unclear, however modulation of its levels by exercise in this model highlights the need for further examination. Overall, our results suggest that exercise might be a beneficial tool that could be used by patients with endometriosis to help control their condition.Support or Funding InformationSupported in part by R15AT009915, R25GM082406, U54MD007579.

Spatial and temporal changes in the expression of steroid hormone receptors in mouse model of endometriosis.

Endometriosis is recognized as a steroid hormone-dependent disorder. However, controversies exist regarding the status of the steroid hormone receptor expression in endometriotic tissues. The purpose of this study was to determine the ontogeny of cellular changes in the expression of estrogen receptors (ERα, ERβ), G protein-coupled estrogen receptor 1 (GPER1), and progesterone receptors (PRs) in endometriosis using a mouse model. We used the autologous uterine tissue transfer mouse model and studied the mRNA and protein expression of ERα, ERβ, GPER1, and PR in ectopic lesions at 2, 4, and 8weeks of induction of endometriosis. As compared to endometrium of controls, in the ectopic endometrium, ERα is reduced while ERβ was elevated in stromal cells; however, Gper1 and PR levels are reduced in both stromal and epithelial cells in a time-specific manner. There is a high inter-animal variation in thelevels of these receptors in ectopic endometrium as compared to controls; the levels also varied by almost 100-fold within the same lesion resulting in "micro-heterogeneity." The expression of all these receptors also deferred between two lesions from the same animal. In the endometriotic tissue, there is extensive inter-animal and intra-lesion heterogeneity in the expression of ERα, ERβ, GPER1, and PR. These changes are not due to the influence of the peritoneal environment but appear to be tissue intrinsic. We propose that the variable outcomes in hormonal therapy for endometriosis could be possibly due to heterogeneity in the expression of steroid hormone receptors in the ectopic endometrium.

Short treatment with antalarmin alters adrenal gland receptors in the rat model of endometriosis.

Endometriosis is a chronic inflammatory disorder in which endometrial tissue is found outside the uterine cavity. Previous reports suggest that there is a dysregulation of the hypothalamic pituitary adrenal axis during the progression of endometriosis. Our previous report showed that a short-term treatment with antalarmin, a corticotrophin releasing hormone receptor type 1 (CRHR1) antagonist decreases the number and size of endometriotic vesicles in the auto-transplantation rat model of endometriosis. Our current goal was to examine the mRNA expression of intra-adrenal receptors to better understand the mechanisms of the hypothalamic pituitary adrenal (HPA) axis involvement in endometriosis. We used two groups of female rats. The first received sham surgery or endometriosis surgery before collecting the adrenals after 7 days of the disease progression. The second group of animals received endometriosis surgery and a treatment of either vehicle or antalarmin (20 mg/kg, i.p.) during the first 7 days after endometriosis induction and then the disease was allowed to progress until day 60. Rats with sham surgery served as controls. Results showed that the mRNA expression of the mineralocorticoid (MRC2) receptor was lower in the rats after 7 days of endometriosis surgery and in rats with endometriosis that received antalarmin. In addition, the CRHR1 was significantly elevated in animals that received antalarmin and this was counteracted by a non-significant elevation in CRHR2 mRNA. The glucocorticoid receptor mRNA within the adrenals was not affected by endometriosis or antalarmin treatment. This report is one of the first to explore intra-adrenal mRNA for receptors involved in the HPA axis signaling as well as in the sympatho-adrenal signaling, calling for additional research towards understanding the role of the adrenal glands in chronic inflammatory diseases such as endometriosis.

Effect of dexamethasone versus propranolol on surgically induced endometriosis in an experimental animal model

Background Endometriosis is a disease of adolescents and reproductive-aged women characterized by the presence of endometrial tissue outside the uterine cavity and commonly associated with chronic pelvic pain and infertility. The mechanisms responsible for its pathogenesis and progression remain poorly understood. It is well established that endometriosis grows and regresses in estrogen-dependent fashion, and the diseases can be effectively cured by definitive surgery. However, prolonged medical therapy may be needed in most of the cases. Objective The objective of this study is to evaluate the effects of dexamethasone versus propranolol in rats with surgically induced endometriosis on vascular endothelial growth factor (VEGF) as a marker in the implanted tissue. Materials and methods The study was conducted in the Animal Research Unit of Medical Research Center of Ain Shams University. All procedures were performed from May 2018 to September 2018, in compliance with the international guidelines for care and use of experimental animals, on 100 female Wistar albino rats 90 days old, which were randomly divided into three groups: first group was treated with dexamethasone (n=40), and second group was treated with propranolol (n=40), and third group was a control group (n=20). They were used to create a model for the experimental induction of endometriosis and evaluation of therapeutic effect of dexamethasone versus propranolol. Results In the first group after treatment with dexamethasone, there was a highly statistically significant decrease in size of implant tissue (P Conclusion Both propranolol and dexamethasone efficiently reduced the size of endometriotic implant in conditions of surgical-induced endometriosis in rats. This is through restriction of angiogenesis through VEGF, which was utilized as a marker. So, it can be deemed that the drugs could be promoted in conditions of endometriosis in humans, but they require more research studies for assessment in humans.