Potential use of ellagic acid for endometriosis treatment: its effect on a human endometrial cell cycle, adhesion and migration

Endometriosis is a common and challenging condition of reproductive-aged women that is defined as the presence of endometrial-like tissue outside the uterine cavity. Despite its prevalence, there is still no effective therapeutics so we aim to evaluate the ellagic acid (EA) effect on the most relevant aspects that are known to be altered in endometriosis. Endometrial primary cultures from women with and without endometriosis and endometrial cell lines were incubated with EA (50 and 100µM) for 24 and 48 h. The results demonstrated that EA arrest endometrial stromal cell cycle on G2 / M phase, after 48 h. In addition, EA 100μM treatment significantly decreased ECC-1 cell migration at 20 h and T-HESC cell migration at 10 h and 20 h; while EA 50μM caused a significant decreased on T-HESC cell migration at 20 h. On the other hand, we proved that treatment with EA for 24 h reduces T-HESC and ECC-1 adhesion to plastic. However, we did not find an effect of EA on cell proliferation. EA has an inhibitory effect on endometrial cell adhesion, migration and cell cycle progression in vitro. These highlight the idea to investigate natural compounds as a novel and promising therapeutic treatment for endometriosis.Fil: Mc Cormack, Bárbara Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Bilotas, Mariela Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Madanes, Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Ricci, Analía Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Singla, José Javier. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; ArgentinaFil: Barañao, Rosa Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

Efecto de los inhibidores de aromatasa en el crecimiento endometrial ectópico y en el medio peritoneal en un modelo murino de endometriosis

El objetivo es valorar los efectos de los inhibidores de aromatasa en los crecimientos endometriales ectópicos y en la liberación de factores angiógenos e inflamatorios al líquido peritoneal (LP). Se realizó un estudio experimental prospectivo en hembras de ratones Balb/c de dos meses de edad. Se practicaron procedimientos quirúrgicos en los ratones para inducir lesiones de tipo endometriósico. Se inició tratamiento con anastrozol o letrozol en los días posoperatorios uno o 28 y se mantuvo durante cuatro semanas. Las lesiones de tipo endometriósico expresaron aromatasa P-450. El tratamiento con anastrozol o letrozol no evitó el desarrollo de las lesiones; sin embargo, sí causó disminución significativa del tamaño de éstas. Cuando se inició el tratamiento con letrozol y anastrozol al primer día posoperatorio, se redujo la proliferación celular y se incrementó la apoptosis; cuando se instituyó en el día 28, ambos inhibidores de aromatasa atenuaron la proliferación celular, pero sólo el anastrozol elevó los niveles de apoptosis. Además, el letrozol redujo las concentraciones de VEGF y PGE en el LP. Con el anastrozol disminuyó el contenido de VEGF, pero no se produjeron cambios significativos de la concentración de PGE.Fil: Bilotas, Mariela Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina; ArgentinaFil: Meresman, Gabriela Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina; ArgentinaFil: Stella, Inés. Hospital Israelita "ezrah"; ArgentinaFil: Sueldo, Carlos. Centro de Estudios En Ginecologia y Reproduccion; ArgentinaFil: Barañao, Rosa Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentina; Argentin

Effect of aromatase inhibitors on ectopic endometrial growth and peritoneal environment in a mouse model of endometriosis

Objective: To evaluate the effect of aromatase inhibitors on ectopic endometrial growth and on the release of proangiogenic and proinflammatory factors in peritoneal fluid (PF). Design: Prospective experimental study. Setting: Animal research and laboratory facility. Animal(s): Female Balb/c mice 2 months of age. Intervention(s): Mice had surgery performed to induce endometriosis-like lesions. Treatment with anastrozole or letrozole was started on either postoperative day 1 or 28 and continued for 4 weeks. Main Outcome Measure(s): Endometriotic lesions were counted and measured and aromatase expression, cell proliferation, and apoptosis were assessed. Vascular endothelial growth factor (VEGF) and prostaglandin E (PGE) levels were evaluated in the PF. Result(s): Endometriosis-like lesions express aromatase P-450. Treatment with either anastrozole or letrozole did not prevent lesion establishment; however, it significantly decreased the size of the endometriotic lesion. When treatment was initiated on postoperative day 1, letrozole and anastrozole decreased cell proliferation and increased apoptosis. When treatment was started on postoperative day 28, both aromatase inhibitors decreased cell proliferation, but only anastrozole augmented apoptosis levels. In addition, letrozole reduced VEGF and PGE levels in PF. Anastrozole diminished VEGF content but did not cause any significant change in PGE levels. Conclusion(s): These findings support the further investigation of aromatase inhibition as a treatment option for endometriosis. © 2010 American Society for Reproductive Medicine.Fil: Bilotas, Mariela Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Meresman, Gabriela Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Stella, Inés. Hospital Israelita; ArgentinaFil: Sueldo, Carlos. Centro de Estudios en Ginecología y Reproducción; ArgentinaFil: Barañao, Rosa Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

PI3K/AKT pathway is altered in the endometriosis patient’s endometrium and presents differences according to severity stage

Based on the inflammatory nature and hormone-dependency of endometriosis, PI3K/AKT signaling appears to influence its progression. Could the endometriosis stages be linked to differential changes in PI3K/AKT pathway regulation? The objective is to evaluate the expression of PI3K, PTEN, AKT and p-AKT in endometrial human biopsies, according to the presence or absence of the disease, and to assess the underlying differences regarding the endometriosis stages. Biopsy specimens of the ectopic and eutopic endometrium were obtained from twenty women with untreated peritoneal endometriosis as well as endometrium biopsies from nine controls. Our study revealed an increased expression of PI3K in eutopic and ectopic endometrium from patients with endometriosis, and a reduced expression of PTEN and increased levels of AKT phosphorylation, compared to control endometrium. Both eutopic and ectopic endometrium from patients with minimal-mild endometriosis expressed a significant reduced PTEN level compared to the respective endometrium from patients with moderate-severe endometriosis. The ratio p-AKT/total AKT showed higher levels of AKT phosphorylation in endometriotic tissue from patients with minimal-mild endometriosis. This study has firmly confirmed the alteration in PI3K/AKT pathway regulation and demonstrated clear differences between the stages of endometriosis, emphasizing the importance of this pathway in the first stage of the disease.基于子宫内膜异位症的炎症性质和激素依赖性, PI3K/AKT信号可能影响其进展。子宫内膜异位症的分期是否与PI3K/AKT通路调节的不同变化有关？本研究目的是根据是否存在子宫内膜异位症, 评估PI3K、PTEN、AKT和p-AKT在子宫内膜活检中的表达, 并评估其子宫内膜异位症分期的潜在差异。对20例未经治疗的腹膜子宫内膜异位症患者的异位和在位子宫内膜进行了活检, 并对9例对照者进行了子宫内膜活检。我们的研究显示, 与对照组相比, 子宫内膜异位症患者在位和异位内膜中PI3K的表达增加, PTEN的表达减少, AKT磷酸化水平升高。轻度子宫内膜异位症患者的在位子宫内膜和异位子宫内膜与中重度子宫内膜异位症患者的相应子宫内膜相比, PTEN水平显著降低, p-AKT/AKT比值显示轻度子宫内膜异位症患者子宫内膜异位组织中AKT磷酸化水平较高。本研究证实了PI3K/AKT通路调节的改变, 并显示了子宫内膜异位症各阶段之间的明显差异, 强调了该通路在疾病第一阶段的重要性。Fil: Madanes, Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Bilotas, Mariela Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Bastón, Juan Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Singla, José Javier. Universidad de Buenos Aires. Facultad de Medicina. Hospital de Clínicas General San Martín; ArgentinaFil: Meresman, Gabriela Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Barañao, Rosa Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Ricci, Analía Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

Effect of pregnancy on apoptosis in endometriotic lesions.

<p>The percentage of apoptotic cells was assessed by TUNEL in epithelial (A) and stromal (B) cells from pregnant (N = 11) and non pregnant (N = 9) mice. Micrographs (C-E) show representative histological sections of endometriotic lesions from pregnant (C) and non pregnant (D) mice. Arrows indicate marked cells. As a negative control, a tissue sample was subjected to treatment without TdT (E). Scale bar indicates 100 μm. Statistical comparisons were performed by Student “t” test. * p<0.05 pregnant vs. non pregnant; *** p<0.001 pregnant vs. non pregnant.</p

Effect of pregnancy and endometriosis on peritoneal IL-2 and IFN-γ.

<p>Mice with surgically induced endometriosis (EDT; N = 20, 11 pregnant) and sham animals (N = 18, 14 pregnant) were mated and sacrificed at day 18 of pregnancy. Levels of IL-2 (A), and IFN-γ (B) were assessed in the peritoneal fluid by ELISA. Statistical comparisons were performed by two way ANOVA test followed by Tukey HSD test for unequal sample size. * p<0.05</p

Effect of pregnancy on cell proliferation in endometriotic lesions.

<p>The percentage of proliferating cells was assessed by immunohistochemistry for PCNA in epithelial (A) and stromal (B) cells from pregnant (N = 11) and non pregnant (N = 9) mice. Micrographs (C-E) show representative histological sections of endometriotic lesions from pregnant (C) and non pregnant (D) mice. Arrows indicate marked cells. As a negative control, immunoglobulin of the same immunoglobulin class and concentration as the primary antibody was used (E). Scale bar indicates 100 μm. Statistical comparisons were performed by Student “t” test. * p<0.05 pregnant vs. non pregnant.</p

Effect of pregnancy on the number and size of endometriotic lesions.

<p>Mice with surgical induced endometriosis were mated and sacrificed at day 18 of pregnancy. The number of endometriotic lesions per mouse (A) and the size of endometriotic lesions (B) in pregnant (N = 11) and non pregnant (N = 9) mice were assessed. Statistical comparisons were performed by Student “t” test. * p<0.05 pregnant vs. non pregnant</p

Effect of pregnancy on endometriotic lesions.

<p><i>Note</i>: Statistical comparisons were performed by Chi-square test. Values are the percentage of lesions that showed signs of leukocyte infiltration, necrosis, decidualization or endometrial involution.</p><p>^a Pregnant vs. non pregnant</p><p>Effect of pregnancy on endometriotic lesions.</p