RESUMO
INTRODUCTION: Endometriosis is a chronic inflammatory disease that affects â¼10 % of women. A significant fraction of patients experience limited or no efficacy with current therapies. Tissue adjacent to endometriosis lesions often exhibits increased neurite and vascular density, suggesting that disease pathology involves neurotrophic activity and angiogenesis. OBJECTIVES: We aim to evaluate the potential for key tyrosine-kinase-receptor-coupled neurotrophic molecules to contribute to endometriosis-associated pain in mice. METHODS: Peritoneal fluid was collected from endometriosis patients undergoing surgery and the levels of NGF and VEGFR1 regulators (VEGFA, VEGFB, PLGF, and sVEGFR1) were quantified by ELISA. VEGFR1 regulator concentrations were used to calculate VEGFR1 occupancy. We used genetic depletion, neutralizing antibodies, and pharmacological approaches to specifically block neurotrophic ligands (NGF or BDNF) or receptors (VEGFR1, TRKs) in a murine model of endometriosis-associated pain. Endometriosis-associated pain was measured using von Frey filaments, quantification of spontaneous abdominal pain-related behavior, and thermal discomfort. Disease parameters were evaluated by lesion size and prevalence. To evaluate potential toxicity, we measured the effect of entrectinib dose and schedule on body weight, liver and kidney function, and bone structure (via micro-CT). RESULTS: We found that entrectinib (pan-Trk inhibitor) or anti-NGF treatments reduced evoked pain, spontaneous pain, and thermal discomfort. In contrast, even though calculated receptor occupancy revealed that VEGFR1 agonist levels are sufficient to support signaling, blocking VEGFR1 via antibody or tamoxifen-induced knockout did not reduce pain or lesion size in mice. Targeting BDNF-TrkB with an anti-BDNF antibody also proved ineffective. Notably, changing dosing schedule to once weekly eliminated entrectinib-induced bone-loss without decreasing efficacy against pain. CONCLUSIONS: This suggests NGF-TrkA signaling, but not BDNF-TrkB or VEGF-VEGFR1, mediates endometriosis-associated pain. Moreover, entrectinib blocks endometriosis-associated pain and reduces lesion sizes. Our results also indicated that entrectinib-like molecules are promising candidates for endometriosis treatment.
RESUMO
Euschistus heros is an important pest in many crops in Brazil, and different control strategies, mainly involving chemicals, have been evaluated; however, the side effects of these chemicals on the balance of inorganic element levels in the hemolymph are unknown. Thus, the aim of this work was to determine the concentration of inorganic elements (focusing on macro-elements) in the hemolymph of female and male E. heros adults, after applying pyriproxyfen at a sublethal concentration (LC30â¯=â¯6.68â¯mLâ¯L-1 diluted in distilled water) to 4th instar nymphs, which were kept in controlled conditions. The hemolymph pool was removed 48â¯h after adult emergence, centrifuged and placed on an acrylic disk added with Gallium as internal standard for the analysis of total reflection X-ray fluorescence. Most of the elements in the control treatment did not differ between females and males. However, following insecticide application to females and males, respectively, there was a significant increase in sulfur (19 and 51%), chlorine (33 and 137%) and calcium (47 and 82%) in the hemolymph. The significantly higher increase in macro-elements in males' hemolymph indicates that the action of pyriproxyfen may be sex-specific. Phosphorus and potassium concentrations also differed between females and males in the control and treated groups. The observed variation in inorganic elements in the insect's hemolymph may be related to the unknown effects of pyriproxyfen, mainly on immune and reproductive performance.