Assessment of the effect of copaiba oil (copaifera sp.) on the kidneys of rats with hepatic dysfunction

Copaiba oil reveals therapeutic properties and has actions such as anti-inflammatory, healing, antiseptic, antitumor, antibacterial, germicidal, expectorant, diuretic and analgesic. Given the anti-inflammatory properties of copaiba oil, the present study aims to evaluate the action of the oil on possible nephrotoxic effects of rats with liver dysfunction caused by the drug thioacetamide. Wistar rats were randomly divided into four groups, consisting of the control group (C), copaiba oil group (O), thioacetamide group (TAA) and thioacetamide + copaiba oil group (TAA + O). The animals received treatment via gavage for eight weeks and were fed standard rodent chow. The kidney fragments were histologically prepared using the historresin technique and HE staining and, later, the images were photographed and processed in an image analyzer for histological characterization of the kidney tissue. The histological results showed an intact kidney tissue with a homogeneous pattern between the groups. Typical cortical and medullary layers and very distinct renal tubules with their well stained and healthy cells. The renal corpuscles showed regular contours, well-distributed anastomosed capillaries and typical macula densa. Histopathological analysis of the kidneys did not reveal alterations or any type of lesion in important structures such as renal tubules and renal corpuscles, and no lymphocytic infiltrate was observed in the renal interstitium. In contrast, the liver tissue from the parallel histological study developed cirrhosis in the group treated with thioacetamide, however, this drug did not compromise the renal histological pattern of the present study. It is concluded that the kidney tissue did not show morphological changes when submitted to thioacetamide, supposedly due to the exposure time, the dose used or that the thioacetamide has been metabolized into less toxic compounds to the kidneys to the detriment of hepatic damage. However, the parallel study of liver tissue and the present study ensure the use of copaiba oil for therapeutic purposes

High Levels of Tumor Necrosis Factor-Alpha Reduce Placental Aquaporin 3 Expression and Impair in vitro Trophoblastic Cell Migration

Placentas from preeclamptic women display augmented tumor necrosis factor-alpha (TNF-α) levels with reduced expression of aquaporin 3 (AQP3). However, whether TNF-α modulates AQP3 expression remains to be elucidated. We hypothesize that elevated levels of TNF-α reduce AQP3 expression and negatively impact trophoblastic cell migration. Spontaneously hypertensive rats (SHRs) and Wistar rats (14–16 weeks) were divided into hypertensive and normotensive groups, respectively. Systolic blood pressure (SBP) was measured, and animals mated. In a third group, pregnant SHRs were treated with a TNF-α antagonist, etanercept (0.8 mg/kg, subcutaneously) on days 0, 6, 12, and 18 of pregnancy. Placentas were collected on the 20th day of pregnancy. Human placental explants, from normotensive pregnancies, were incubated with TNF-α (5, 10, and 20 ng/ml) and/or etanercept (1 μg/ml). Swan 71 cells were incubated with TNF-α (10 ng/ml) and/or etanercept (1 μg/ml) and subjected to the wound healing assay. AQP3 expression was assessed by Western blot and TNF-α levels by ELISA. SBP (mmHg) was elevated in the hypertensive group, and etanercept treatment reduced this parameter. Placental TNF-α levels (pg/ml) were higher in the hypertensive group. AQP3 expression was reduced in the hypertensive group, and etanercept treatment reversed this parameter. Explants submitted to TNF-α exposition displayed reduced expression of AQP3, and etanercept incubation reversed it. Trophoblastic cells incubated with TNF-α showed decreased cell migration and reduced AQP3 expression, and etanercept incubation ameliorated it. Altogether, these data demonstrate that high TNF-α levels negatively modulate AQP3 in placental tissue, impairing cell migration, and its relationship in a pregnancy affected by hypertension.Fil: Rodrigues Dos Passos Junior, Rinaldo. Universidade Federal de Goiás; BrasilFil: Alves de Freitas, Raiany. Universidade Federal de Goiás; BrasilFil: Reppetti, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Medina Mora, Yollyseth Astrid. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Dela Justina, Vanessa. Universidade Federal de Goiás; BrasilFil: Werle Bach, Camila. Universidade Federal do Mato Grosso do Sul; BrasilFil: Facholi Bomfim, Gisele. Universidade Federal do Mato Grosso do Sul; BrasilFil: Vitorino Lima, Victor. Universidade Federal do Mato Grosso do Sul; BrasilFil: Damiano, Alicia Ermelinda. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; ArgentinaFil: Gianchini, Fernanda. Universidade Federal de Goiás; Brasi

Óleo-resina de copaíba diminui o índice de adiposidade e melhora o sistema redox, os níveis de IL-10 e função renal de ratos submetidos à dieta rica em sacarose

Alterações nos hábitos alimentares levam a maior risco de desenvolver obesidade e outras doenças crônicas, como disfunção renal. Produtos naturais podem apresentar potencial terapêutico, sendo importante avaliar os efeitos desses compostos. Objetivou-se avaliar o efeito do óleo-resina de copaíba sobre os rins de ratos submetidos à dieta rica em sacarose. Para isso, ratos Wistar machos foram casualmente divididos para receberem dieta padrão e solução de sacarose (30%) (S, n=8) ou dieta padrão e solução de sacarose (30%) e suplementação com óleo-resina de copaíba (200 mg/kg/dia, via gavagem) (S+OC, n=8), por 8 semanas. Ao final do experimento, foram avaliados ganho de peso, índice de adiposidade, peso dos rins, marcadores do estado redox e níveis de citocinas. A análise estatística foi realizada por teste t de Student, com nível de significância de P<0,05. Foi observado que a suplementação com óleo-resina de copaíba foi eficiente em reduzir o ganho de peso e índice de adiposidade. Não houve diferença no peso dos rins entre os grupos. A suplementação com óleo-resina não alterou os níveis de ureia, contudo, diminuiu os níveis de creatinina, foi capaz de aumentar os níveis de IL-10, a atividade das enzimas CAT e SOD, e reduzir os marcadores de dano oxidativo. Assim, conclui-se que o óleo-resina de copaíba reduz o índice de adiposidade, o qual foi associado a melhora do estado redox, ao aumento de IL-10 e à melhor função renal, sugerindo que o óleo-resina de copaíba apresenta efeitos benéficos sobre os rins de animais submetidos à dieta rica em sacarose

Role of Toll-like receptor 4 in vascular reactivity in spontaneously hypertensive rats.

Nosso objetivo foi verificar a participação do TLR4 na pressão arterial e reatividade vascular em ratos SHR. O TLR4 está mais expresso em artérias mesentéricas de resistência de SHR com 15 semanas do que em Wistar com 15 e SHR com 5 semanas. SHR e Wistar com 15 semanas foram tratados com anti-TLR4 (1mg/dia) ou IgG (controle) por 15 dias via i.p. A expressão do TLR4, MyD88, a fosforilação da P38 e NF-kB p65, e a secreção de IL-6 foi menor nos SHR anti-TLR4 do que nos SHR IgG. Os SHR tratados com anti-TLR4 apresentaram redução na pressão arterial versus SHR IgG. A resposta máxima ao KCl e à noradrenalina (NA) foram normalizadas após o uso do anti-TLR4 no SHR, por vias dependentes de COX-1, COX-2 e do NF-kB. O uso do L-NAME diminuiu a resposta contrátil à NA no SHR IgG sendo que o anti-TLR4 melhorou essa resposta em SHR. O anti-TLR4 aumentou a expressão da eNOS e preveniu a geração de espécies reativas de oxigênio em SHR. Sugerimos que o TLR4 está associado com o aumento da pressão arterial e com a disfunção vascular presente na hipertensão arterial.Our objective was to investigate the role of TLR4 in blood pressure and vascular reactivity in SHR. TLR4 is more expressed in mesenteric resistance arteries from SHR 15 week than in Wistar with 15 and SHR 5 weeks. Wistar and SHR with 15 weeks were treated with anti-TLR4 (1mg/dia) or IgG (control) for 15 days via ip. The expression of TLR4, MyD88, phosphorylation of p38 and NF-kB p65, and IL-6 secretion was lower in SHR TLR4 than in SHR IgG. SHR treated with anti-TLR4 had reduced blood pressure versus SHR IgG. The maximal response to KCl and noradrenaline (NA) were normalized after anti-TLR4 treatment in SHR by mechanisms dependent on COX-1, COX-2 and NF-kB. The use of L-NAME decreased the contractile response to NA in SHR IgG and anti-TLR4 improved this response in SHR. The anti-TLR4 increased eNOS expression and prevented the generation of reactive oxygen species in SHR. We suggest that the TLR4 is associated with increased blood pressure and vascular dysfunction in hypertension

Monocyte-derived dendritic cells from patients with dermatophytosis restrict the growth of Trichophyton rubrum and induce CD4-T cell activation.

Dermatophytes are the most common agents of superficial mycoses that are caused by mold fungi. Trichophyton rubrum is the most common pathogen causing dermatophytosis. The immunology of dermatophytosis is currently poorly understood. Recently, our group investigated the interaction of T. rubrum conidia with peritoneal mouse macrophages. We found that macrophages phagocytose T. rubrum conidia resulted in a down-modulation of class II major histocompatibility complex (MHC) antigens and in the expression of co-stimulatory molecules. Furthermore, it induced the production of IL-10, and T. rubrum conidia differentiated into hyphae that grew and killed the macrophages after 8 hrs of culture. This work demonstrated that dendritic cells (DCs) and macrophages, from patients or normal individuals, avidly interact with pathogenic fungus T. rubrum. The dermatophyte has two major receptors on human monocyte-derived DC: DC-SIGN and mannose receptor. In contrast macrophage has only mannose receptor that participates in the phagocytosis or bound process. Another striking aspect of this study is that unlike macrophages that permit rapid growth of T. rubrum, human DC inhibited the growth and induces Th activation. The ability of DC from patients to interact and kill T. rubrum and to present Ags to T cells suggests that DC may play an important role in the host response to T. rubrum infection by coordinating the development of cellular immune response

Outcome of <i>T.rubrum</i> infection in macrophages and dendritic cells.

<p>Cells layered on coverslips inserted in 24-well tissue culture microplates and incubated with conidia for different time intervals. (a) The coverslips were washed and stained with Giemsa for micrographs. (A) Macrophages and dendritic cells cultured without conidia for 10 h; (B), macrophage infected with conidia for 4 h; (C) 8 h and (D) 12 h. (b) Fungal citotoxicity was determined by LDH assay. The arrow indicates the presence of conidea. *p<0.05 as compared with macrophages and dendritic cell cultivated with <i>T.rubrum</i> conidia after 4 h of interaction.</p

Phagocytic index of <i>T. rubrum</i> in presence inhibitors of mannose and DC-SIGN receptors or both.

<p>Macrophages and dendritic cells was incubated with 1, 10, 20 and 50 µg/ml of antibody and co-cultured with <i>T. rubrum</i> conidia at a ratio of 1∶3. After 12 h the cells were fixed and stained with Giemsa. Internalized fungal cells were visualized by light microscopy and the phagocytic index determined (A). As control was used the purified mouse IgG1 kappa isotype to anti-CD206, and purified mouse IgG2b kappa isotype control to anti-CD209. The graph represents the inhibition using 10 µg/ml of antiboby, that was determined previously as the optimal dose (A). Effect of exoantigen (B) on phagocytosis of <i>T. rubrum</i> conidia by macrophages and dendritic cells. Macrophages and dendritic cells were incubated with exoantigen at different concentrations for 30 min before the addition of <i>T. rubrum.</i> *p<0.05 when compared with macrophages or dendritic cells cultivated with isotype antibody or unstimulated (control).</p

Cytokine production by CD4+ T-cell in presence of DCs and <i>T. rubrum</i> conidia (rate conidia/cell = 3∶1).

<p>After 5 days of proliferation the supernatant was harvested and cytokine determined. The results represent the mean ± SD of three independents experiments, where the blood was collected from each study subjects (10 patients and 10 controls). As negative control was used only CD4+ T-cell and the result of cytokine production always <20 pg/ml. Each dot correspond to an individual. *p<0.05 was considerate significant.</p

Influence of <i>T. rubrum</i> conidia on the release of TNF-α, IL-12 and IL-10 by dendritic cells (A) and macrophages (B).

<p>After 12 h the supernatants were harvested, then TNF-α, IL-12 and IL-10 quantified by ELISA. Results are representative of three independent experiments. Each dot correspond to an individual. #p<0.05 when compared with cells unstimulated, *p<0.05 when compared with macrophages.</p

Proliferation of CD4+ T-cell in presence of <i>T. rubrum</i> conidea-pulsed DCs from patients or non-infected individuals.

<p>On day 7, DCs were pulsed with or without conidia of <i>T.rubrum</i> (rate conidia/cell = 3∶1) by 24 hr. After, pulsed DCs were plated at 2×10⁴ in triplicate wells of a 96-well flat-bottom plate. Autologous CD4+ T-cell were added at 2×10⁵ cells/well. Proliferation of CD4 T-cell was evaluated after 5 days by measuring [³H]thymidine uptake and results were expressed as counts per minutes (CPM). The results represent the mean ± SD of three independents experiments, where the blood was collected from each study subjects (10 patients and 10 controls). As negative control was used only CD4+ T-cell and the result of proliferation was <100 cpm. Each dot correspond to an individual. *p<0.05 was considerate significant.</p