Helicteres sacarolha A. St.- Hil. et al.: gastroprotective and possible mechanism of actions in experimental animals

AbstractEthnopharmacological relevanceHelicteres sacarolha A. St.- Hil. et al. popularly known in Brazil as ‘semente-de-macaco’, is widely employed in the popular medicine in many of parts of Brazil in the alleviation of symptoms of ailments such as peptic ulcer and inflammation. Up to the present, there is no study addressing the gastroprotective activity of the hydroethanolic extract of H. sacarolha and its possible mechanism of actions.Materials and methodsThe hydroethanolic (70%) extract of H. sacarolha (HEHs) was obtained by maceration. The gastroprotective activity was assessed using gastric ulcer models induced by acidified ethanol, piroxicam, and water restraint stress in mice and rats at doses of 20, 50 and 250mg/kg p.o. Mechanistic studies involved the antisecretory assay evaluated with pylorus ligation in rats and pre-treatments with appropriate antagonists/inhibitors such as yohimbine, glibenclamide, indomethacin and l-NAME, effect on catalase and myeloperoxidase activities and gastric mucus determination using acidified ethanol- induced ulcer in mice.ResultsHEHs at all doses tested demonstrated potent gastroprotective activities in the acute ulcer models. The gastroprotective activity of HEHs was attenuated by pre-treatments with yohimbine, glibenclamide, indomethacin and l-NAME. HEHs effectively reduced basal gastric juice production without any effect on the free and total acidity. The gastroprotective action of HEHs involved increasing the antioxidant enzyme catalase and mucus secretion and inhibition of neutrophyl infiltration as reflected by the reduction in the myeloperoxidase activity.ConclusionThe results of this study gave a scientific support for the popular use of the leaves of H. sacarolha in the treatment of gastric ulcers and that it has a multi-targeted action

Differential regulation of the release of tumor necrosis factor-alpha and of eicosanoids by mast cells in rat airways after antigen challenge.

BACKGROUND: Rat trachea display a differential topographical distribution of connective tissue mast cells (CTMC) and mucosal mast cells (MMC) that may imply regional differences in the release of allergic mediators such as tumor necrosis factor-alpha (TNF-alpha) and eicosanoids. AIM: To evaluate the role of CTMC and MMC for release of TNF-alpha and eicosanoids after allergenic challenge in distinct segments of rat trachea. MATERIALS AND METHODS: Proximal trachea (PT) and distal trachea (DT) from ovalbumin (OVA)-sensitized rats, treated or not with compound 48/80 (48/80) or dexamethasone, were incubated in culture medium. After OVA challenge, aliquots were collected to study release of TNF-alpha and eicosanoids. RESULTS: Release of TNF-alpha by PT upon OVA challenge peaked at 90 min and decayed at 6 and 24 h. Release from DT peaked at 30-90 min and decayed 6 and 24 h later. When CTMC were depleted with 48/80, OVA challenge exacerbated the TNF-alpha release by PT at all time intervals, while DT exacerbated TNF-alpha levels 6 and 24 h later only. Dexamethasone reduced TNF-alpha production after 90 min of OVA challenge in PT and at 3 and 6h in DT. OVA challenge increased prostaglandin D2) in DT and leukotriene B4 in both segments but did not modify prostaglandin E2 and leukotriene C4 release. CONCLUSION: OVA challenge induces TNF-alpha release from MMC, which is negatively regulated by CTMC. The profile of TNF-alpha and eicosanoids depends on the time after OVA challenge and of the tracheal segment considered

Roux-en-y gastric bypass improves in short term the clinical-anthropometric parameters and reduces risk for obesity-related cardiometabolic diseases / Bypass gástrico roux-en-y melhora a curto prazo os parâmetros clínico-antropométricos e reduz o risco de doenças cardiometabólicas relacionadas à obesidade

Roux-en-Y gastric bypass surgery (RYGB) is the most applied technique in the treatment of severe obesity worldwide. However, its impact on anthropometric parameters and the risk for cardiometabolic diseases in obese patients is uncertain. To evaluate anthropometric clinical parameters and the evolution of risk factors for obesity-related diseases in individuals of both sexes undergoing RYGB. Sixty-nine adults subjects from both sexes submitted to RYGB surgery treatment were divided into 3 groups: G1(<13 months, n=24); G2 (>13 and <25 months, n=21), and G3 (>25 and <37 months, n=24). Sociodemographic and anthropometric information before and after surgery were collected. The abdominal perimeter was used in the classification of cardiometabolic risk and the BMI was used for the risk of obesity-related diseases. Hypotheses were tested by Student's t-test and ANOVA, and the significance level adopted was 5%. The average age was 36.0±10.0 years, with 69.6% being male and 30.4% female. Anthropometric parameters (weight, BMI, and abdominal circumference) were higher among women, except for weight loss and percentage of weight loss. There was a difference in weight loss between the sexes in the moments before and after RYGB. There was a decrease in the risk of disease due to obesity and cardiovascular diseases after RYGB. Weight loss and %WL were greater years by year in the short term of 3 years after surgery. RYGB proved to be an effective strategy for both sexes in combating obesity, providing in the short term a significant improvement in clinical-anthropometric parameters and reduction of risk factors for obesity-related cardiometabolic diseases

The putative role of ovary removal and progesterone when considering the effect of formaldehyde exposure on lung inflammation induced by ovalbumin

OBJECTIVE: Formaldehyde exposure during the menstrual cycle is known to affect the course of allergic lung inflammation. Because our previous data demonstrated that formaldehyde combined with an ovariectomy reduced allergic lung inflammation, we investigated the putative role of ovary removal and progesterone treatment when considering the effect of formaldehyde on allergic lung inflammation. METHOD: Ovariectomized rats and their matched controls were exposed to formaldehyde (1%, 3 days, 90 min/ day) or vehicle, and immediately after exposure, the rats were sensitized to ovalbumin by a subcutaneous route. After 1 week, the rats received a booster by the same route, and after an additional week, the rats were challenged with ovalbumin (1%) by an aerosol route. The leukocyte numbers, interleukin-10 (IL-10) release, myeloperoxidase activity, vascular permeability, ex vivo tracheal reactivity to methacholine and mast cell degranulation were determined 24 h later. RESULTS: Our results showed that previous exposure to formaldehyde in allergic rats decreased lung cell recruitment, tracheal reactivity, myeloperoxidase activity, vascular permeability and mast cell degranulation while increasing IL-10 levels. Ovariectomy only caused an additional reduction in tracheal reactivity without changing the other parameters studied. Progesterone treatment reversed the effects of formaldehyde exposure on ex vivo tracheal reactivity, cell influx into the lungs and mast cell degranulation. CONCLUSION: In conclusion, our study revealed that formaldehyde and ovariectomy downregulated allergic lung inflammation by IL-10 release and mast cell degranulation. Progesterone treatment increased eosinophil recruitment and mast cell degranulation, which in turn may be responsible for tracheal hyperreactivity and allergic lung inflammation

Female sex hormones mediate the allergic lung reaction by regulating the release of inflammatory mediators and the expression of lung E-selectin in rats

Abstract\ud \ud Background\ud Fluctuations of estradiol and progesterone levels caused by the menstrual cycle worsen asthma symptoms. Conflicting data are reported in literature regarding pro and anti-inflammatory properties of estradiol and progesterone.\ud \ud \ud Methods\ud Female Wistar rats were ovalbumin (OVA) sensitized 1 day after resection of the ovaries (OVx). Control group consisted of sensitized-rats with intact ovaries (Sham-OVx). Allergic challenge was performed by aerosol (OVA 1%, 15 min) two weeks later. Twenty four hours after challenge, BAL, bone marrow and total blood cells were counted. Lung tissues were used as explants, for expontaneous cytokine secretion in vitro or for immunostaining of E-selectin.\ud \ud \ud Results\ud We observed an exacerbated cell recruitment into the lungs of OVx rats, reduced blood leukocytes counting and increased the number of bone marrow cells. Estradiol-treated OVx allergic rats reduced, and those treated with progesterone increased, respectively, the number of cells in the BAL and bone marrow. Lungs of OVx allergic rats significantly increased the E-selectin expression, an effect prevented by estradiol but not by progesterone treatment. Systemically, estradiol treatment increased the number of peripheral blood leukocytes in OVx allergic rats when compared to non treated-OVx allergic rats. Cultured-BAL cells of OVx allergic rats released elevated amounts of LTB4 and nitrites while bone marrow cells increased the release of TNF-α and nitrites. Estradiol treatment of OVx allergic rats was associated with a decreased release of TNF-α, IL-10, LTB4 and nitrites by bone marrow cells incubates. In contrast, estradiol caused an increase in IL-10 and NO release by cultured-BAL cells. Progesterone significantly increased TNF- α by cultured BAL cells and bone marrow cells.\ud \ud \ud Conclusions\ud Data presented here suggest that upon hormonal oscillations the immune sensitization might trigger an allergic lung inflammation whose phenotype is under control of estradiol. Our data could contribute to the understanding of the protective role of estradiol in some cases of asthma symptoms in fertile ans post-menopausal women clinically observed.The authors gratefully acknowledge Dr. Gabriela Cavriani for her help in this\ud study and Zilma Lucia da Silva (Depth of Pharmacology) of Institute of\ud Biomedical Sciences of University of São Paulo (São Paulo, Brazil) for\ud technical assistance and for Mayara Munhóz de Assis Ramos and Suzanne\ud Kane of Los Angeles, California for further English revisions to our\ud manuscript. This study was supported by Fundação de Amparo à Pesquisa\ud do Estado de São Paulo (FAPESP) Grants 2001/13384-4, 2004/14128-0, 2006/\ud 55950-0, 2006/14128-4, 2007/55631-4, 2009/51886-3 and 2009/07208-0 and\ud CAPES (PNPD 0188085, 02610/09-4). W. Tavares de Lima is a fellow\ud researcher of CNPq

Long-term amphetamine treatment exacerbates inflammatory lung reaction while decreases airway hyper-responsiveness after allergic stimulus in rats

Asthma is an allergic lung disease can be modulated by drugs that modify the activity of central nervous system (CNS) such as amphetamine (AMPH). AMPH is a highly abused drug that exerts potent effects on behavior and immunity. In this study we investigated the mechanism involved in the effects of long-term AMPH treatment on the increased magnitude of allergic lung response. We evaluated mast cells degranulation, cytokines release, airways responsiveness and, expression of adhesion molecules. Male Wistar rats were treated with AMPH or vehicle (PBS) for 21 days and sensitized with ovalbumin (OVA) one week after the first injection of vehicle or AMPH. Fourteen days after the sensitization, the rats were challenged with an OVA aerosol, and 24 h later their parameters were analyzed. In allergic rats, the treatment with AMPH exacerbated the lung cell recruitment due increased expression of ICAM-1, PECAM-1 and Mac-1 in granulocytes and macrophages recovered from bronchoalveolar lavage. Elevated levels of IL-4, but decreased levels of IL-10 were also found in samples of lung explants after AMPH treatment. Conversely, the ex-vivo tracheal hyper-responsiveness to methacholine (MCh) was reduced by AMPH treatment, whereas the force contraction of tracheal segments due to in vitro antigen challenge remained unaltered. Our findings suggest that lung inflammation and airway hyper-responsiveness due to OVA challenge are under the distinct control of AMPH during long-term treatment. Our data strongly indicate that AMPH positively modulates allergic lung inflammation via the increase of ICAM-1, PECAM-1, Mac-1 and IL-4. AMPH also abrogates the release of the anti-inflammatory cytokine IL-10. (c) 2012 Elsevier B.V. All rights reserved.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2007/55631-4, 2009/51886-3, 2009/07208-0, 2008/50766-1]CNPq [300764/2010-3]CAPES [02610/09-4

Annexin 1 genic and proteic analysis in the developmental biology and in the pathophysiology of the inflammation

The annexin 1 (ANXA1) is a 37 kDa protein that has been implicated in the regulation of phagocytosis, cell signaling, apoptosis and leukocyte transmigration. Originally described as a phospholipase A2-inhibitory protein, ANXA1 can regulate several components of the inflammatory reaction, such as cytokines. Recently, a line of animals lacking the AnxA1 gene was generated with dual purpose targeting construct, designed simultaneously to inactivate the AnxA1 gene and to report upon its activity using LacZ reporter gene. In the developmental biology, the epithelial and leukocytes, studied in different organs, display AnxA1 gene expression. In the liver, the hepatocytes synthesize this protein during the embriogenesis, disappearing in adults. In those animals, the ANXA1 is re-synthesized during cancer, regeneration or inflammatory process. In the intramembranous ossification, the densitometric and histological analysis of newborn ANXA1 null skull bones showed that the absence of the ANXA1 induced a delay in this process. In models of experimental acute inflammation, ANXA1 null mice exhibited an exaggerated response and a partial or complete resistance to the anti-inflammatory effects of glucocorticoids. Several other anomalies were noted, including increased spontaneous migratory behavior of leukocytes and mast cell degranulation. Also, the absence of ANXA1 during the acute inflammation induce a disregulation in the leukocyte expression of adhesion molecules, such as selectin CD26L and integrin CD11b, leading to a higher cell adhesion in the ANXA1 null mice. During the endotoxemic response, the ANXA1 null mice exhibited a toxic response characterized by a marked degree of leukocyte adhesion and an aberrant expression of Toll-like receptor 4 in macrophages. These alterations have resulted in organ injury and lethality within 48 hours, a phenotype rescued by exogenous administration of ANXA1. Finally, the pleiotropic and pluripotent role of ANXA1 may be explained by the phosphorylation of its N-terminal domain, which induces specific actions, for example the regulation of cell growth, the vesicles aggregation, and the translocation of ANXA1 to cell membrane. In conclusion, the study of ANXA1 genic and proteic activities in the developmental biology or in the homeostasis of the inflammatory response might design the development of novel anti-inflammatory therapeutics based on this endogenous mediator.A anexina 1 (ANXA1) é uma proteína de 37 kDa, associada com a regulação dos processos de fagocitose, sinalização celular, apoptose e migração leucocitária. Originalmente descrita como uma proteína inibidora da citosólica fosfolipase A2, a ANXA1 pode regular vários componentes da reação inflamatória, tais como as citocinas. Recentemente, uma linhagem de camundongos deficientes para a ANXA1 foi gerada com a finalidade de, simultaneamente, inativar o gene da AnxA1 e analisar sua atividade a partir da expressão do gene da LacZ. Na biologia do desenvolvimento, as células epiteliais e leucócitos, estudados nos diversos órgãos, expressam o gene da AnxA1. No fígado, os hepatócitos sintetizam essa proteína apenas durante a embriogênese, desaparecendo nos animais adultos. Nestes animais, a ANXA1 reaparece nos processos de regeneração, tumorais e inflamatórios. Na ossificação intramembranosa, as análises de densitometria óssea e histológica dos ossos do crânio dos camundongos ANXA1 null recém-nascidos indicaram que a ausência da ANXA1 induz um retardo neste processo. Nos modelos de inflamação aguda, os animais ANXA1 null exibiram uma resposta exacerbada e uma resistência parcial ou completa aos efeitos antiinflamatórios dos glicocorticóides. Além disso, outras anormalidades foram observadas nos animais deficientes, incluindo um aumento da migração leucocitária e do processo de desgranulação dos mastócitos. Ainda, na inflamação aguda, a ausência da ANXA1 desregula a expressão de moléculas de adesão dos leucócitos, como a selectina CD26L e a integrina CD11b, induzindo um aumento na adesão destas células em animais ANXA1 null. Na investigação realizada durante a endotoxemia experimental, os camundongos ANXA1 null desenvolveram uma resposta tóxica caracterizada pelo aumento da adesão dos leucócitos e pela expressão anormal de Toll-like receptor 4 nos macrófagos. A ocorrência destas alterações resultou em injúria tecidual e letalidade em 48 horas, patologia que foi revertida com a administração da ANXA1. Finalmente, o papel pleiotrópico e pluripotente da ANXA1 pode ser explicado pela fosforilação da região N-terminal, induzindo ações específicas, como a regulação no crescimento celular, agregação de vesículas e translocação da ANXA1 para a superfície celular. Concluindo, o estudo da atividade gênica e protéica da ANXA1 na biologia do desenvolvimento ou na homeostase dos processos inflamatórios poderá definir o desenvolvimento de novas terapias antiinflamatórias baseadas neste mediador endógeno.TEDEBV UNIFESP: Teses e dissertaçõe

Cellular aspects and pharmacological mediators envolved in acute and chronic inflamatory reponse

BV UNIFESP: Teses e dissertaçõe

Effect of annexin-A1 peptide treatment during lung inflammation induced by lipopolysaccharide

Lung endotoxemia is characterized by neutrophil accumulation, increased vascular permeability and parenchymal injury. This can also affect the endogenous pathways that operate in the host to keep inflammation under control. Here, we demonstrate differential expression of annexin-A1 (AnxA1) protein in mice after the local or intraperitoneal administration of lipopolysaccharide (LPS; 1 mg/kg) in mice and the regulation of the endotoxemic inflammation after the pre-treatment with the AnxA1 peptidomimetic Ac2-26. The intranasal administration of LPS induced the leukocyte migration and cytokine release to the alveolar space, whereas the peritoneal administration of LPS generated a deregulated cellular and cytokine response, with a marked degree of leukocyte adhesion in the microcirculation. The peptide Ac2-26 pre-treatment inhibited the leukocyte migration and the pro-inflammatory cytokine release. Also, it induced the expression of endogenous AnxA1 and the antiinflammatory cytokine IL-10. In conclusion, our data obtained from endotoxemia induced by local or intraperitoneal LPS administration suggested that the molecular mechanisms induced by AnxAl peptidomimetic Ac2-26 lead to the regulation of leukocyte activation/migration and cytokine production induced by LPS. (C) 2012 Elsevier Ltd. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq