Bisphenol A disruption promotes mammary tumor microenvironment via phenotypic cell polarization and inflammatory response.

Inflammation in the established tumor microenvironment (TME) is often associated with a poor prognosis of breast cancer. Bisphenol A (BPA) is an endocrine-disrupting chemical that acts as inflammatory promoter and tumoral facilitator in mammary tissue. Previous studies demonstrated the onset of mammary carcinogenesis at aging when BPA exposure occurred in windows of development/susceptibility. We aim to investigate the inflammatory repercussions of BPA in TME in mammary gland (MG) during neoplastic development in aging. Female Mongolian gerbils were exposed to low (50 µg/kg) or high BPA (5000 µg/kg) doses during pregnancy and lactation. They were euthanized at 18 months of age (aging) and the MG were collected for inflammatory markers and histopathological analysis. Contrarily to control MG, BPA induced carcinogenic development mediated by COX-2 and p-STAT3 expression. BPA was also able to promote macrophage and mast cell (MC) polarization in tumoral phenotype, evidenced by pathways for recruitment and activation of these inflammatory cells and tissue invasiveness triggered by tumor necrosis factor-alpha and transforming growth factor-beta 1 (TGF-ß1). Increase of tumor-associated macrophages, M1 (CD68 + iNOS+) and M2 (CD163+) expressing pro-tumoral mediators and metalloproteases was observed; this aspect greatly contributed to stromal remodeling and invasion of neoplastic cells. In addition, the MC population drastically increased in BPA-exposed MG. Tryptase-positive MCs increased in disrupted MG and expressed TGF-ß1, contributing to EMT process during carcinogenesis mediated by BPA. BPA exposure interfered in inflammatory response by releasing and enhancing the expression of mediators that contribute to tumor growth and recruitment of inflammatory cells that promote a malignant profile.

Effect of piperlongumine during exposure to cigarette smoke reduces inflammation and lung injury.

Chronic obstructive pulmonary disease (COPD) is related to smoking and anti-inflammatory therapy is indicated. Among the mediators with anti-inflammatory properties, we highlight piperlongumine (PL), an alkaloid/amide of Piper longum. Here we evaluated the PL administration on an experimental model of respiratory inflammation resulting from exposure to cigarette smoke. Male Balb/c mice were exposed to burning of 10 commercial cigarettes, 2x/day, for five weeks on specific equipment. PL efficacy was evaluated in control, exposed to smoke without treatment and PL treated (2.0 mg/kg, 3x/week) groups. Animals were weighed and plethysmographic analyses performed at the end of the exposure protocol. Inflammatory cells were evaluated in the bronchoalveolar lavage (BAL) and hemoglobin and glucose in the blood. Lung fragments were processed for histopathological studies and AnxA1, COX-2, NF-kB and neutrophil elastase expressions. Plethysmography revealed that PL maintained pulmonary frequency, volume and ventilation parameters similar to controls, with respiratory volume reduction compared to untreated animals. Final weight was reduced in both exposed groups. PL decreased hemoglobin concentration, attenuated the reduction of glucose levels and reduced influx of lymphocytes, neutrophils and macrophages in BAL. Histopathologically occured infiltration of inflammatory cells, increase of the interalveolar septa and intra-alveolar spaces in untreated animals. But, PL administration recovered lung tissues and, immunohistochemically, promoted increased expression of AnxA1 and reduction of COX-2, NF-kB and neutrophil elastase. Together the results indicate that PL attenuates systemic and pulmonary inflammatory changes, partially by modulating the expression the endogenous AnxA1, and may represent a promising therapy in preventing the inflammation induced by cigarette smoke.

Expression of the Annexin A1 and its correlation with matrix metalloproteinases and the receptor for formylated peptide-2 in diffuse astrocytic tumors.

Astrocytomas represent the majority of cerebral gliomas. Studies show that the anti-inflammatory protein Annexin-A1 (ANXA1) is associated with the tumor invasion process and that its actions can be mediated by the receptor for formylated peptides (FPR). Therefore, we evaluated the expression of ANXA1, the receptor FPR2 and matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) in brain astrocytomas. Detection of proteins was performed in sections of diffuse astrocytomas (grade II), anaplastic astrocytomas (grade III) and glioblastomas (GBM, grade IV) and quantifications were made by densitometry. Our analyses showed increased expression of ANXA1 in astrocytomas of all grades, but especially in GBM. The expression of FPR2 is similar to that found for ANXA1, being higher in GBM. Immunostaining for MMPs is also stronger as the degree of malignancy increases, especially with respect to MMP-9. The positive correlation between ANXA1/FPR2 and ANXA1/MMP-9 was observed in all tumors studied. The data indicate the possible action of ANXA1 and FPR2 on the development and progression of astrocytomas, related to increased expression of MMP-9. Thereby, ANXA1 and FPR2 are involved in the biology and malignancy of diffuse astrocytic tumors.

Protective effects of the galectin-1 protein on in vivo and in vitro models of ocular inflammation.

PURPOSE: Galectin-1 (Gal-1) is a ß-galactoside-binding protein with diverse biological activities in the pathogenesis of inflammation but has been poorly investigated in terms of ocular inflammation. In the present study, we monitored the anti-inflammatory effects of Gal-1 using the in vivo rodent model of endotoxin-induced uveitis (EIU) and in vitro assays with human RPE (ARPE-19) cells. METHODS: For this purpose, EIU was induced by subcutaneous sterile saline injection of 0.1 ml of lipopolysaccharide (LPS, 1 mg/Kg) in the rat paw, which was maintained under these conditions for 24 h. The therapeutic efficacy of recombinant Gal-1 (rGal-1) was tested in the EIU animals by intraperitoneal inoculation (3 µg/100 µl per animal) 15 min after the LPS injection. In vitro studies were performed using LPS-stimulated ARPE-19 cells (10 µg/ml) for 2, 8, 24 and 48 h, treated or not with rGal-1 (4 µg/ml) or dexamethasone (Dex, 1.0 µM). RESULTS: Gal-1 treatment attenuated the histopathological manifestation of EIU via the inhibition of polymorphonuclear cells (PMN) infiltration in the eye and by causing an imbalance in adhesion molecule expression and suppressing interleukin (IL)-1ß, IL-6, and monocyte chemotactic protein-1 (MCP-1) productions. Immunohistochemical and western blotting analyses revealed significant upregulation of Gal-1 in the eyes induced by EIU after 24 h. In the retina, there was no difference in the Gal-1 expression, which was high in all groups, demonstrating its structural role in this region. To better understand the effects of Gal-1 in the retina, in vitro studies were performed using ARPE-19 cells. Ultrastructural immunocytochemical analyses showed decreased levels of endogenous Gal-1 in LPS-stimulated cells (24 h), while Dex treatment upregulated this protein. The protective effects of rGal-1 on LPS-stimulated cells were associated with the significant reduction of the release of cytokines (IL-8 and IL-6), similar to Dex treatment. Furthermore, rGal-1 and Dex inhibited cyclooxygenase-2 (COX-2) expression in LPS-stimulated cells, as shown by immunofluorescence. CONCLUSIONS: Overall, this study identified potential roles for Gal-1 in ocular inflammation, especially uveitis, and may lead to future therapeutic approaches.

Morphological variations on vital systems in a conjoined twin.

Conjoined twins (CTs), popularly referred to as Siamese twins, are a rare anomaly due to monochorionic and monoamniotic twin pregnancies. Dicephalus dibrachius dipus, a type of parapagus conjoined twin which is characterized by possessing two arms, two legs, a single trunk and two heads, epidemiologically, is an even rarer occurrence of CTs. In this article, a rare, well-preserved anatomical specimen of a dicephalus dibrachius dipus conjoined twin is presented. This study was conducted in a specimen which is part of the collection of the Embryology Museum of the institution by donation and approved by the Research Ethics Committee (REC). The female conjoined twins were born at full-term by cesarean section in the 1970s and died hours after birth. A thorough anatomical description was made through observational analysis, computed tomography and 3D reconstructed images. Major abnormalities were observed in the cardiovascular, respiratory and digestive systems. The internal anatomy exhibited a heart with three atria, two ventricles, two aortic arches, two pulmonary arteries, one innominate venous trunk and a respiratory system with two tracheas and four lungs. No other report was similar to our three atria heart description. This article provides a thorough anatomical description of all systems, which is valuable information for further studies on CTs.

The impact of endogenous annexin A1 on glucocorticoid control of inflammatory arthritis.

OBJECTIVES: To establish the role and effect of glucocorticoids and the endogenous annexin A1 (AnxA1) pathway in inflammatory arthritis. METHODS: Ankle joint mRNA and protein expression of AnxA1 and its receptors were analysed in naive and arthritic mice by real-time PCR and immunohistochemistry. Inflammatory arthritis was induced with the K/BxN arthritogenic serum in AnxA1(+/+) and AnxA1(-/-) mice; in some experiments, animals were treated with dexamethasone (Dex) or with human recombinant AnxA1 or a protease-resistant mutant (termed SuperAnxA1). Readouts were arthritic score, disease incidence, paw oedema and histopathology, together with pro-inflammatory gene expression. RESULTS: All elements of the AnxA1 pathway could be detected in naive joints, with augmentation during ongoing disease, due to the infiltration of immune cells. No difference in arthritis intensity of profile could be observed between AnxA1(+/+) and AnxA1(-/-) mice. Treatment of mice with Dex (10 µg intraperitoneally daily from day 2) afforded potent antiarthritic effects highly attenuated in the knockouts: macroscopic changes were mirrored by histopathological findings and pro-inflammatory gene (eg, Nos2) expression. Presence of proteinase 3 mRNA in the arthritic joints led the authors to test AnxA1 and the mutant SuperAnxA1 (1 µg intraperitoneally daily in both cases from day 2), with the latter one being able to accelerate the resolving phase of the disease. CONCLUSION: AnxA1 is an endogenous determinant for the therapeutic efficacy of Dex in inflammatory arthritis. Such an effect can be partially mimicked by application of SuperAnxA1 which may represent the starting point for novel antiarthritic therapeutic strategies.

Exercise Training and Verbena officinalis L. Affect Pre-Clinical and Histological Parameters.

Verbena officinalis L. or vervain is an herbal medicine and dietary supplement used worldwide. It is used for antidepressant and anticonvulsant purposes, as well as to treat inflammatory disorders, skin burns, abrasions, and gastric diseases, among others. Here, we investigated the biochemical, antioxidant, and histopathological effects of vervain against chronic physical stress. Male Wistar rats were submitted to chronic physical training and oral administration of 200 mg/kg of extract for 7 weeks. Control animals were not treated with either stress or vervain. Body weight was monitored during the study. Liver, kidney, spleen, testis, epididymis, heart, skeletal muscle, and brain samples were collected. Blood cholesterol, lactate dehydrogenase (LDH), bilirubin, and creatinine kinase (CREA), among others, were studied. Glutathione peroxidase (GPox) and superoxide dismutase (SOD) antioxidant activity was analyzed in the blood, liver, and kidney. Testosterone measurements were also performed on whole testis extracts. We found significant weight ratios differences in the epididymis, brain, and heart. Animals submitted to training showed hemorrhagic livers. Kidney histology was affected by both stress and vervain. Cell disruption and vacuolization were observed in the testes and epididymis of animals submitted to stress. Hematological and biochemical markers as CREA, LDH, TP, CKI, URCA, γGT, and glucose revealed statistically significantly differences. Additionally, the activity of glutathione peroxide (GPox) and superoxide dismutase (SOD) in the blood was also impacted. Both stress and vervain have significant in vivo effects. Infusions of vervain include phenylpropanoids, iridoids, verbenalin, hastatoside, and flavonoids, amongst others, which interact synergistically to produce the preclinical effects reported here.

Evaluation of the healing properties of Garcinia brasiliensis extracts in a cutaneous wound model.

ETHNOPHARMACOLOGICAL RELEVANCE: Wound healing is a complex process that can leave pathological scars, especially in case of infections from opportunistic microorganisms. In this context, herbal medicines open up great possibilities for investigation. One of the species of interest native to Brazil is Garcinia brasiliensis ("bacupari"). Traditionally known for treating wounds and ulcers, G. brasiliensis presents anti-inflammatory, antioxidant and antimicrobials properties. But, its wound healing profile in experimental models, in order to validate its efficacy, is still litle studied. AIM OF THE STUDY: Thus, the objective of this work was to evaluate, in an infected cutanous wound model, the potential of formulations incorporated with G. brasiliensis leaves extracts. MATERIALS AND METHODS: Crude extract (CE), Ethyl Acetate Fraction (EAF) and Hexanic Fraction (HF) were submitted to phytochemical assays, high performance thin layer chromatography (HTPLC) and cytotoxicity studies. CE and EAF were also tested for microbicidal properties and incorporated in cream and gel formulations at 10% concentration. After stability testing, the gel formulations with CE or EAF at 10% were selected and applied to skin wounds infected or not with Staphylococcus aureus in Wistar rats. The healing potenttial of the extracts was verified by the expression of the protein Annexin A1 (AnxA1), related to the processes of inflammation and antifibrotic function, the cells immunostaining for Gasdermin-D (GSDM-D), a marker of pyroptotic cell death, and the dosage of interleukin-10 (IL-10) and monocyte chemotactic protein (MCP)-1 inflammatory mediators. RESULTS: Phytochemical studies indicated the presence of compounds of pharmacological interest, including Catechin, Quercetin and Berberine in addition to low cytotoxicity of CE and EAF at 10%. After the 6-day topical treatments, CE and EAF gel formulations demonstrated to control the pruritus formation process. The treatments decreased AnxA1 expression and the amount of cells immunostained for GSDM-D, and increased the expression of MCP-1 in infected wounds. CONCLUSIONS: Together, the results show important anti-inflammatory profile and skin healing potential of CE and EAF from G. brasiliensis leaves, even in infected lesions, with therapeutic perspectives.

Mast cells modulate the inflammatory process in endotoxin-induced uveitis.

PURPOSE: To investigate the role of mast cells and annexin-A1 (Anxa1) in endotoxin-induced uveitis (EIU). METHODS: EIU was induced by injection of lipopolysaccharide (LPS) into the paws of rats, which were then sacrificed after 24 and 48 h. To assess EIU in the absence of mast cells, groups of animals were pretreated with compound 48/80 (c48/80) and sacrificed after 24 h after no treatment or EIU induction. The eyes were used for histological studies and the aqueous humor (AqH) pool was used for the analysis of transmigrated cells and Anxa1 levels. In inflammatory cells, Anxa1 expression was monitored by immunohistochemistry. RESULTS: After 24 h, rats with EIU exhibited degranulated mast cells, associated with elevated numbers of infiltrating leukocytes and the high expression of Anxa1 in the AqH and the neutrophils. After 48 h of EIU, the mast cells were intact, indicating granule re-synthesis, and there was a reduction of neutrophil transmigration and an increase in the number of mononuclear phagocytic cells in ocular tissues. Anxa1 expression was decreased in neutrophils but increased in mononuclear phagocytic cells. In the animals pretreated with c48/80 and subjected to EIU, mast cells responded to this secretagogue by degranulating and few transmigrated neutrophils were observed. CONCLUSTIONS: We report that mast cells are a potential source of pharmacological mediators that are strongly linked to the pathophysiology of EIU, and the endogenous protein Anxa1 is a mediator in the homeostasis of the inflammatory process with anti-migratory effects on leukocytes, which supports further studies of this protein as an innovative therapy for uveitis.

Annexin A1 Mimetic Peptide and Piperlongumine: Anti-Inflammatory Profiles in Endotoxin-Induced Uveitis.

Uveitis is one of the main causes of blindness worldwide, and therapeutic alternatives are worthy of study. We investigated the effects of piperlongumine (PL) and/or annexin A1 (AnxA1) mimetic peptide Ac2-26 on endotoxin-induced uveitis (EIU). Rats were inoculated with lipopolysaccharide (LPS) and intraperitoneally treated with Ac2-26 (200 µg), PL (200 and 400 µg), or Ac2-26 + PL after 15 min. Then, 24 h after LPS inoculation, leukocytes in aqueous humor, mononuclear cells, AnxA1, formyl peptide receptor (fpr)1, fpr2, and cyclooxygenase (COX)-2 were evaluated in the ocular tissues, along with inflammatory mediators in the blood and macerated supernatant. Decreased leukocyte influx, levels of inflammatory mediators, and COX-2 expression confirmed the anti-inflammatory actions of the peptide and pointed to the protective effects of PL at higher dosage. However, when PL and Ac2-26 were administered in combination, the inflammatory potential was lost. AnxA1 expression was elevated among groups treated with PL or Ac2-26 + PL but reduced after treatment with Ac2-26. Fpr2 expression was increased only in untreated EIU and Ac2-26 groups. The interaction between Ac2-26 and PL negatively affected the anti-inflammatory action of Ac2-26 or PL. We emphasize that the anti-inflammatory effects of PL can be used as a therapeutic strategy to protect against uveitis.

Protective effects of piperlongumin in the prevention of inflammatory damage caused by pulmonary exposure to benzopyrene carcinogen.

Benzopyrene is one of the main polycyclic aromatic hydrocarbons with carcinogenic capacity. Research has shown that anti-inflammatory drugs can reduce the incidence of lung cancer. In this scenario, we highlight piperlongumin (PL), an alkaloid from Piper longum with anti-inflammatory properties. Therefore, our aim was to study the effect of PL administration in a model of pulmonary carcinogenesis induced by benzopyrene in Balb/c mice. Animals were divided into 3 groups (n = 10/group): sham (10% DMSO), induced by benzopyrene (100 mg/kg, diluted in DMSO) without treatment (BaP) for 12 weeks and induced by benzopyrene and treated with PL (BaP/PL) (2 mg/kg in 10% DMSO) from the eighth week post-induction. Animals were weighed daily and pletsmography was performed in the 12th week. Genotoxicity and hemoglobin levels were analyzed in blood and quantification of leukocytes in bronchoalveolar lavage (BAL). Lungs were collected for histopathological evaluation, immunohistochemical studies of annexin A1 (AnxA1), cyclooxygenase 2 (COX-2), anti-apoptotic protein Bcl-2 and nuclear transcription factor (NF-kB) and also the measurement of interleukin cytokines (IL)-1ß, IL-17 and tumor necrosis factor (TNF) -α. Treatment with PL reduced the pulmonary parameters (p < 0,001) of frequency, volume and pulmonary ventilation, decreased lymphocytes, monocytes and neutrophils in BAL (p < 0,05) as well as blood hemoglobin levels (p < 0,01). PL administration also reduced DNA damage and preserved the pulmonary architecture compared to the BaP group. Moreover, the anti-inflammatory effect of PL was evidenced by the maintenance of AnxA1 levels, reduction of COX-2 (p < 0,05), Bcl-2 (p < 0,01) and NF-kB (p < 0,001) expressions and decreased IL-1ß, IL-17 (p < 0,01) and TNF-α (p < 0,05) levels. The results show the therapeutic potential of PL in the treatment of pulmonary anti-inflammatory and anti-tumor diseases with promising therapeutic implications.

Anti-Acanthamoeba synergistic effect of chlorhexidine and Garcinia mangostana extract or α-mangostin against Acanthamoeba triangularis trophozoite and cyst forms.

Acanthamoeba spp. can cause amoebic keratitis (AK). Chlorhexidine is effective for AK treatment as monotherapy, but with a relative failure on drug bioavailability in the deep corneal stroma. The combination of chlorhexidine and propamidine isethionate is recommended in the current AK treatment. However, the effectiveness of treatment depends on the parasite and virulence strains. This study aims to determine the potential of Garcinia mangostana pericarp extract and α-mangostin against Acanthamoeba triangularis, as well as the combination with chlorhexidine in the treatment of Acanthamoeba infection. The minimal inhibitory concentrations (MICs) of the extract and α-mangostin were assessed in trophozoites with 0.25 and 0.5 mg/mL, for cysts with 4 and 1 mg/mL, respectively. The MIC of the extract and α-mangostin inhibited the growth of A. triangularis trophozoites and cysts for up to 72 h. The extract and α-mangostin combined with chlorhexidine demonstrated good synergism, resulting in a reduction of 1/4-1/16 of the MIC. The SEM results showed that Acanthamoeba cells treated with a single drug and its combination caused damage to the cell membrane and irregular cell shapes. A good combination displayed by the extract or α-mangostin and chlorhexidine, described for the first time. Therefore, this approach is promising as an alternative method for the management of Acanthamoeba infection in the future.

VEGFA and NFE2L2 Gene Expression and Regulation by MicroRNAs in Thyroid Papillary Cancer and Colloid Goiter.

Deregulation of VEGFA (Vascular Endothelial Growth Factor A) and NFE2L2 (Nuclear Factor (Erythroid-derived 2)-Like 2), involved in angiogenesis and oxidative stress, can lead to thyroid cancer progression. MiR-17-5p and miR-612 are possible regulators of these genes and may promote thyroid disorders. In order to evaluate the involvement of VEGFA, NFE2L2, hsa-miR-17-5p, and hsa-miR-612 in thyroid pathology, we examined tissue samples from colloid goiter, papillary thyroid cancer (PTC), and a normal thyroid. We found higher levels of VEGFA and NFE2L2 transcripts and the VEGFA protein in goiter and PTC samples than in normal tissue. In the goiter, miR-612 and miR-17-5p levels were lower than those in PTC. Tumors, despite showing lower VEGFA mRNA expression, presented higher VEGFA protein levels compared to goiter tissue. In addition, NRF2 (Nuclear Related Transcription Factor 2) protein levels in tumors were higher than those in goiter and normal tissues. Inhibition of miR-17-5p resulted in reduced NFE2L2 expression. Overall, both transcript and protein levels of NFE2L2 and VEGFA were elevated in PTC and colloid goiter. Hsa-miR-612 showed differential expression in PTC and colloid goiter, while hsa-miR-17-5p showed differential expression only in colloid goiter, suggesting that hsa-miR-17-5p may be a positive regulator of NFE2L2 expression in PTC.

Modulation of the endogenous Annexin A1 in a cigarette smoke cessation model: Potential therapeutic target in reversing the damage caused by smoking?

BACKGROUND: Smoking cessation may help in the reversal of inflammation and damage caused by smoking. The endogenous annexin A1 (AnxA1) protein has anti-inflammatory effects which instigates the understanding of its role in the attenuation of inflammatory processes caused by smoking. MATERIAL AND METHODS: Wistar rats were exposed to cigarette smoke for 8 weeks. After the exposure period, one of the groups remained other 8 weeks in the absence of smoke. Animals not exposed to smoke were used as control. Blood, trachea and lungs were obtained for histopathological, immunohistochemical and biochemical analyses. RESULTS: Loss of cilia of the tracheal lining epithelium was found by smoke exposure, but smoking cessation led to recovery of the tracheal epithelium. Similarly, chronically exposed-to-smoke animals showed increased lymphocytes and macrophages in bronchoalveolar lavage and higher levels of glucose and gamma-GT in their blood. Reduction of lymphocytes, glucose and gamma-GT occurred after smoking cessation. In addition, IL-1ß, IL-6, IL-10, TNF-α and MCP-1 levels were elevated by smoke exposure. Smoking cessation significantly reduced the levels of IL-1ß, IL-6 and MCP-1 but increased the IL-10 concentration. Numerous mast cells and macrophages were observed in the lung of chronically exposed-to-smoke animals with reduction by smoking cigarette abstinence. AnxA1 increased expression and concomitant NF-κB reduction were found in the smoking cessation group. CONCLUSION: Our results showed that cigarette abstinence promoted partial recovery of the inflammatory process. The attenuation of the inflammatory profile may be associated with the overexpression of AnxA1 protein.

Salicornia ramosissima J. Woods seeds affected the normal regenerative function on carbon tetrachloride-induced liver and kidney injury.

The growing importance of Salicornia plants as bioactive agents and health promoters associated with the continuous demand for alternative treatments for liver disorders, has stimulated us to evaluate the renal and hepatic effects of S. ramosissima seeds in mice under normal conditions and exposure to toxic products as carbon tetrachloride (CCl4). Thus, histopathological and lipid peroxidation evaluations of the liver and kidneys were performed. Powdered dried seeds of S. ramosissima (SRS) were administered orally for 22 days at a dose of 2000 mg/kg/day to male mice in three different settings: 1) seed effects, 2) protection against CCl4 acute toxicity (0.2 mL/kg) and 3) regeneration after acute exposure to CCl4 (0.2 mL/kg), each study being performed with appropriate control animals. Mice treated with SRS per se had slightly enlarged hepatic sinusoids and noticeable renal inflammation. SRS did not show effective protection against mice exposed to CCl4 and had no positive influence on liver and kidney recovery after CCl4 administration. These results demonstrated that SRS failed to improve hepato- and nephrotoxicity, in addition to the apparent synergism between CCl4 and SRS under these experimental conditions. Although the biological mechanisms of S. ramosissima are not fully understood, the evidence suggests further research to elucidate its adverse biological effects.

Mimetic peptide AC2-26 of annexin A1 as a potential therapeutic agent to treat COPD.

Chronic obstructive pulmonary disease (COPD) is related to inflammatory process caused by smoking habit. In this scenario, the anti-inflammatory protein Annexin A1 (AnxA1) may represent a therapeutic alternative. We performed experiments to evaluate the effects of the AnxA1 mimetic peptide Ac2-26 in an initial COPD model by physiological, histopathological, biochemical and immunohistochemical analyses. Weight loss, increased blood pressure, reductions in the pulmonary frequency and ventilation, loss of tracheal cilia, enlargement of the pulmonary intra-alveolar spaces and lymphoid tissue found in untreated smoke-exposed group were attenuated by AnxA1 peptide treatment. The Ac2-26 administration also protected against leukocytes influx in bronchoalveolar lavage (BAL), lung and trachea, and it also led to decreased hemoglobin, glucose, cholesterol, gamma glutamyl transferase and aspartato aminotransferase levels. Similarly, reduction of proinflammatory mediators and higher concentration of anti-inflammatory cytokine were found in macerated lung supernatant, blood plasma and BAL in the treated animals. Besides Ac2-26 group showed reduced tissue expressions of AnxA1, cyclooxygenase-2 and metalloproteinase-9, but formylated peptide receptor 2 (FPR2) overexpression. Our results all together highlighted the protective role of the Ac2-26 mimetic peptide in COPD with promising perspectives.

Efeitos anti-inflamatórios do extrato alcoólico de folhas de garcinia gardneriana na peritonite induzida por lipopolissacarídeo / Anti-inflammatory effects of garcinia gardneriana leaf extract on lipopolysaccharideinduced peritonitis

Introdução: A Garcinia gardneriana é utilizada na medicina tradicional brasileira para o tratamento de tumores, inflamações e alívio de dores, mas as informações científicas são ainda limitadas. Objetivos: Diante do uso popular e o anseio por efeitos colaterais mínimos, o objetivo geral deste estudo foi avaliar as propriedades anti-inflamatórias da G. gardneriana em modelo de peritonite induzido por lipopolissacarideo (LPS). Métodos: Ratos Wistar foram divididos aleatoriamente em 3 grupos (n= 5/ grupo): controle, induzido à peritonite e não tratado e induzido à peritonite e tratado com extrato de folhas alcoólico de G. gardneriana a 4%. A peritonite foi induzida por única injeção intraperitoneal de LPS (1 mg/kg). O tratamento com o extrato foi realizado por gavagem (1 ml), administrado antes e 12h após a injeção do LPS. Os ratos foram eutanasiados após 24h da indução de peritonite. Amostras de sangue foram coletadas para análise plasmática de histamina, o lavado intraperitoneal para quantificação de neutrófilos e o intestino delgado para processamento histológico, quantificação de mastócitos e imuno-histoquímica da expressão da proteína Anexina A1 (AnxA1). Resultados: As análises quantitativas indicaram os efeitos anti-inflamatórios do extrato, pela redução do recrutamento de neutrófilos para a cavidade peritoneal e a diminuição da quantidade de mastócitos na lâmina própria do intestino delgado, comparadas aos animais não tratados. Não houve diferença estatística dos níveis de histamina. A imuno-histoquímica indicou diminuição acentuada da expressão da AnxA1 na mucosa intestinal dos animais tratados. Conclusão: Nossos dados demonstraram que o extrato alcoólico de G. gardneriana tem forte ação anti-inflamatória e potencial terapêutico para o desenvolvimento de fitoterápicos com propriedades anti-inflamatórias

Introduction: Garcinia gardneriana is used in traditional Brazilian medicine for the treatment of tumors, inflammation and relief of pain, but scientific information is still limited. Objective: In the face of popular use and the desire for minimal side effects, the general objective of this study was to evaluate the anti-inflammatory properties of G. gardneriana in a model of lipopolysaccharide-induced peritonitis (LPS). Methods: Wistar rats were randomly divided into 3 groups (n = 5 / group): control, induced to peritonitis and untreated and induced to peritonitis and treated with 4% alcoholic extract of G. gardneriana leaves. Peritonitis was induced by single intraperitoneal injection of LPS (1 mg/kg). Treatment with the extract was performed by gavage (1 ml), given before and 12h after LPS injection. The rats were euthanized 24h after the peritonitis induction. Blood samples were collected for plasma analysis of histamine, intraperitoneal lavage for quantification of neutrophils and the small intestine for histological processing for quantification of mast cells, and immunohistochemical analysis of the expression of Annexin A1 (AnxA1) protein. Results: Quantification analyses indicated the anti-inflammatory effects of the extract by reducing the recruitment of neutrophils into the peritoneal cavity and reducing the amount of mast cells in the lamina propria of the small intestine compared to the untreated animals. There was no statistical difference in the levels of histamine. Immunohistochemical studies indicated a marked decrease of the AnxA1 expression in the intestinal mucosa of the treated animals. Conclusion: Our data demonstrated that the alcoholic extract of G. gardneriana has a strong anti-inflammatory action and therapeutic potential for the development of herbal products with anti-inflammatory properties

Introducción: Garcinia gardneriana se utiliza en la medicina tradicional brasileña para el tratamiento de tumores, inflamaciones y alivio del dolor, pero la información científica aún es limitada. Objetivo: Frente al uso popular y la búsqueda de efectos secundarios mínimos, lo objetivo general de este estudio fue evaluar las propiedades antiinflamatorias de G. gardneriana en un modelo de peritonitis inducido por lipopolisacárido (LPS). Métodos: Se dividieron aleatoriamente ratas Wistar en 3 grupos (n= 5/grupo): control, inducido a peritonitis y no tratado, e inducido a peritonitis y tratado con extracto alcohólico de hojas de G. gardneriana al 4%. La peritonitis fue inducida por una única inyección intraperitoneal de LPS (1 mg/kg). El tratamiento con el extracto se realizó por gavaje (1 ml), administrado antes y 12 horas después de la inyección de LPS. Las ratas fueron sacrificadas después de 24 horas de la inducción de peritonitis. Se recopilaron muestras de sangre para el análisis plasmático de histamina, el lavado intraperitoneal para la cuantificación de neutrófilos y el intestino delgado para el procesamiento histológico, la cuantificación de mastocitos y la inmunohistoquímica de la expresión de la proteína Anexina A1 (AnxA1). Resultados: Los análisis cuantitativos indicaron los efectos antiinflamatorios del extracto, mediante la reducción del reclutamiento de neutrófilos en la cavidad peritoneal y la disminución de la cantidad de mastocitos en la lámina propia del intestino delgado, en comparación con los animales no tratados. No hubo diferencia estadística en los niveles de histamina. La inmunohistoquímica indicó una disminución pronunciada de la expresión de AnxA1 en la mucosa intestinal de los animales tratados. Conclusión: Nuestros datos demostraron que el extracto alcohólico de G. gardneriana tiene una fuerte acción antiinflamatoria y potencial terapéutico para el desarrollo de fitoterapéuticos con propiedades antiinflamatorias.

Anti-inflammatory actions of herbal medicines in a model of chronic obstructive pulmonary disease induced by cigarette smoke.

The effects of four medicinal herbs (Arctium lappa, Plantago major, Mikania glomerata Spreng and Equisetum arvense) with anti-inflammatory properties were evaluated in a chronic obstructive pulmonary disease (COPD) model. Wistar rats were exposed to cigarette smoke during 8 weeks and one of the groups was orally given a solution containing 4% of each alcoholic herbal extracts during the exposure period. Control group was not exposed to smoke or treated. Histopathological, immunohistochemical and biochemical analyzes were performed. Normal blood plasma levels of gamma glutamyl transferase indicated no toxicity of the administered herbal extracts. The treatment reduced leukocytes influx in bronchoalveolar lavage, mast cell and macrophages numbers in lungs, as well as prevented pulmonary congestion and tracheal metaplasia. Herbal mixture also decreased plasma inflammatory mediator levels and pulmonary expression of annexin A1 and nuclear factor-k?. Our data indicate synergistic and protective effects of the used herbal medicines in animals exposed to cigarette smoke as a potential therapeutic strategy.

Heterogeneity of mast cells and expression of Annexin A1 protein in a second degree burn model with silver sulfadiazine treatment.

Mast cells (MCs) participate in all stages of skin healing and one of their mediators is the Annexin A1 protein (AnxA1), linked to inflammation, proliferation, migration and apoptosis processes, but not studied in thermal burns yet. Therefore, our objectives were to evaluate the behavior of MCs and AnxA1 in a second degree burn model, treated or not with silver sulfadiazine 1% (SDP 1%) and associated to macrophages quantification and cytokines dosages. MCs counts showed few cells in the early stages of repair but increased MCs in the final phases in the untreated group. The normal skin presented numerous tryptase-positive MCs that were reduced after burning in all analyzed periods. Differently, few chymase-positive MCs were observed in the early stages of healing, however, increased chymase-positive MCs were found at the final phase in the untreated group. MCs also showed high immunoreactivity for AnxA1 on day 3 in both groups. In the tissue there was a strong protein expression in the early stages of healing, but in the final phases only in the SDP treated animals. TNF-α, IL-1ß, IL-6, IL-10 and MCP-1 levels and macrophages quantification were increased in inflammation and reepithelialization phases. Reduced IL-1ß, IL-6 and IL-10 levels and numerous macrophages occurred in the treated animals during tissue repair. Our results indicate modulation in the profile of MCs and AnxA1expression during healing by the treatment with SDP 1%, pointing them as targets for therapeutic interventions on skin burns.

Avaliação dos perfis fitoquímicos, antioxidante e citotóxicos do extrato alcoólico de folhas de cajá-manga (spondias dulcis) / Evaluation of phytochemical, antioxidant and cytotoxic profiles of the alcoholic extract of cajamanga leaves (spondias dulcis) / Evaluación de perfiles fitoquímicos, antioxidante y citotóxicos del extracto alcohólico de hojas de cajamanga (spondias dulcis)

Introdução: As espécies de plantas frutíferas, além da importância nutricional, podem ser usadas como fontes de compostos bioativos de interesse para o desenvolvimento de novos medicamentos. Entre as plantas com potencial terapêutico destacamos a Spondias dulcis (cajá-manga) indicada pela medicina tradicional para tratamento de condições médicas dos sistemas tegumentar, respiratório e genitourinário. Objetivos: Realizar a caraterização fitoquímica, antioxidante e citotóxica do extrato alcoólico bruto de folhas de Spondia dulcis. Material e Métodos: O extrato bruto foi obtido por percolação com o uso de 20 g das folhas secas e trituradas de Spondia dulcis e 100 ml de etanol a 70o por 24h. Na padronização do extrato foram utilizadas diferentes reações para análises de identificação de compostos do metabolismo secundário, bem como a determinação da atividade antioxidante pela captura do radical livre do DPPH. Após retirada do álcool por rotaevaporação, foram realizados testes de citotoxidade in vitro (hemólise) em solução glicosilada (5%) de hemácias (4%) nas diferentes concentrações do extrato (0,5%, 1% e 1,5%). Resultados: As análises fitoquímicas qualitativas identificaram a presença de compostos fenólicos, flavonoides, taninos genéricos e cumarinas. A avaliação dos compostos terpenoides mostrou presença de saponinas e ausência de sesquiterpenos e triterpenos. Os alcaloides foram detectados pelas reações de Bouchardat, Dragendorff, Mayer e Sheibler. A análise da atividade antioxidante do extrato indicou alta capacidade antioxidante (71%). No ensaio de hemólise a citotoxidade foi baixa nas concentrações de 0,5% e 1% e relativa na concentração de 1,5%. Conclusão: O extrato alcoólico bruto de Spondia dulciscontém compostos bioativos anti-inflamatórios, alta capacidade antioxidante e baixa citotoxicidade até a concentração de 1,5% com potencial de aplicação farmacológica (AU)

Introduction: Fruit plant species, in addition to their nutritional importance, can be used as sources of bioactive compounds of interest for the development of new drugs. Among the plants with therapeutic potential, we highlight Spondias dulcis (cajá-manga) indicated by traditional medicine for the treatment of medical conditions of the integumentary, respiratory and genitourinary systems. Objectives: To carry out the phytochemical, antioxidant and cytotoxic characterization of the leaves crude alcoholic extract of Spondia dulcis. Material and Methods: The crude extract was obtained by percolation using 20 g of dried and crushed leaves of Spondia dulcis and 100 ml of ethanol at 70° for 24 hours. In the standardization of the extract, different reactions were used to analyze the identification of compounds of secondary metabolism, as well as the determination of the antioxidant activity by capturing the free radical of DPPH. After removing the alcohol by rotary evaporation, in vitro cytotoxicity tests (hemolysis) were performed in a glycosylated solution (5%) of red blood cells (4%) at different concentrations of the extract (0.5%, 1% and 1.5%). Results: Qualitative phytochemical analyzes identified the presence of phenolic compounds, flavonoids, generic tannins and coumarins. The evaluation of terpenoid compounds showed the presence of saponins and the absence of sesquiterpenes and triterpenes. Alkaloids were detected by Bouchardat, Dragendorff, Mayer and Sheibler reactions. The analysis of the antioxidant activity of the extract indicated that the crude extract of Spondia dulcis has a high antioxidant capacity (71%). In the hemolysis assay, cytotoxicity was low at concentrations of 0.5% and 1% and relative at concentrations of 1.5%. Conclusion: The crude alcoholic extract of Spondia dulcis contains anti-inflammatory bioactive compounds, high antioxidant capacity and low cytotoxicity up to a concentration of 1.5% with potentialfor pharmacological application (AU)

Introducción: Las especies de plantas frutales, además de su importancia nutricional, pueden ser utilizadas como fuentes de compuestos bioactivos de interés para el desarrollo de nuevos fármacos. Entre las plantas con potencial terapéutico destacamos Spondias dulcis (cajamanga) indicada por la medicina tradicional para el tratamiento de afecciones médicas de los sistemas tegumentario, respiratorio y genitourinario. Objetivos: Realizar la caracterización fitoquímica, antioxidante y citotóxica del extracto alcohólico crudo de hojas de Spondia dulcis. Material y Métodos: El extracto crudo se obtuvo por percolación utilizando 20 g de hojas secas y trituradas de Spondia dulcis y 100 ml de etanol a 70° durante 24 horas. En la estandarización del extracto se utilizaron diferentes reacciones para analizar la identificación de compuestos de metabolismo secundario, así como la determinación de la actividad antioxidante mediante la captura del radical libre de DPPH. Después de eliminar el alcohol por evaporación rotatoria, se realizaron pruebas de citotoxicidad (hemólisis) in vitro en una solución glicosilada (5%) de glóbulos rojos (4%) a diferentes concentraciones del extracto (0,5%, 1% y 1,5%). Resultados: Los análisis fitoquímicos cualitativos identificaron la presencia de compuestos fenólicos, flavonoides, taninos genéricos y cumarinas. La evaluación de los compuestos terpenoides mostró la presencia de saponinas y la ausencia de sesquiterpenos y triterpenos. Los alcaloides fueron detectados por las reacciones de Bouchardat, Dragendorff, Mayer y Sheibler. Los estudios han demostrado que el extracto crudo de Spondiadulcis tiene una alta capacidad antioxidante (71%). En el ensayo de hemólisis, la citotoxicidad fue baja a concentraciones de 0,5% y 1% y relativa a concentraciones de 1,5%. Conclusión: El extracto alcohólico crudo de Spondia dulcis contiene compuestos bioactivos antiinflamatorios, alta capacidad antioxidante y baja citotoxicidad hasta una concentración de 1,5% conpotencial de aplicación farmacológica (AU)