Tissue healing changes on wounds in rats after treatment with Hancornia speciosa latex in cream-gel formulation.

PURPOSE: Hancornia speciosa latex has shown pharmacological potential in wound healing processes due to its angiogenic, osteogenic, and anti-inflammatory activities. The aims of this study were to carry out a cream-gel formulation with 5, 10 and 25% of H. speciosa serum latex and to evaluate its potential to stimulate the skin regeneration in rats' wounds. METHODS: One hundred and twenty rats were divided into five groups: neutral control with saline (G1), cream-gel based on H. speciosa latex serum at 5% m/v (G2), cream-gel at 15% m/v (G3), cream-gel at 25% m/v (G4), and cream-gel (G5). The animals were euthanized at three, seven, 14 and 21 days after the injury induction, and some parameters were analyzed: wound contraction, necrosis, fibrin, polymorphonuclear and mononuclear infiltrates, fibroblast, angiogenesis, hemorrhage, and collagen. RESULTS: The therapeutic treatment with cream-gel at 15 and 25% is beneficial in the inflammatory phase of healing processes since it increased the angiogenesis and proliferation of mononuclear infiltrations in wounds. Regarding wound contraction, the treatment with cream-gel (5 and 15%) induced a higher rate of contraction in the proliferative phase. The 15% cream-gel formulation stimulated a greater production of collagen in the injured tissues. CONCLUSIONS: H. speciosa cream-gel is a low-cost herbal medicine which can aid in tissue repair.

Ru(II)/amino acid complexes inhibit the progression of breast cancer cells through multiple mechanism-induced apoptosis.

For some cancer subtypes, such as triple-negative breast cancer, there are no specific therapies, which leads to a poor prognosis associated with invasion and metastases. Ruthenium complexes have been developed to act in all steps of tumor growth and its progression. In this study, we investigated the effects of Ruthenium (II) complexes coupled to the amino acids methionine (RuMet) and tryptophan (RuTrp) on the induction of cell death, clonogenic survival ability, inhibition of angiogenesis, and migration of MDA-MB-231 cells (human triple-negative breast cancer). The study also demonstrated that the RuMet and RuTrp complexes induce cell cycle blockage and apoptosis of MDA-MB-231 cells, as evidenced by an increase in the number of Annexin V-positive cells, p53 phosphorylation, caspase 3 activation, and poly(ADP-ribose) polymerase cleavage. Moreover, morphological changes and loss of mitochondrial membrane potential were detected. The RuMet and RuTrp complexes induced DNA damage probably due to reactive oxygen species production related to mitochondrial membrane depolarization. Therefore, the RuMet and RuTrp complexes acted directly on breast tumor cells, leading to cell death and inhibiting their metastatic potential; this reveals the potential therapeutic action of these drugs.

Hancornia speciosa serum fraction latex stimulates the angiogenesis and extracellular matrix remodeling processes.

The Hancornia speciosa latex reveals angiogenic, osteogenic, and anti-inflammatory properties, which present its potential for developing of wound healing drugs; however, the latex compounds responsible for angiogenesis remain unknown. One strategy to screen these active compounds is evaluation of latex fractions. This study aimed to obtain different fractions of latex and evaluate its angiogenic activity separately using the chick chorioallantoic membrane (CAM) assay. The serum (SE) fraction was responsible for angiogenesis, which was subject to biochemical characterization and computational simulations in order to understand the contribution of H. speciosa latex in wound healing process. Our results revealed weak antioxidant potential and absence of antimicrobial activity in the SE fraction. Phytochemical analysis identified chlorogenic acids (CGA) as the main compound of SE fraction. CGA bioactivity predictions identify different molecules associated with extracellular matrix (ECM) remodeling, such as metalloproteinases, which also are overexpressed in our CAM assay experiment. Docking simulations revealed the interactions between CGA and matrix metalloproteinase 2. In conclusion, SE latex fraction stimulates angiogenesis and may influence ECM remodeling. These properties may contribute to the wound healing process, and also confirm the widespread use of this plant.

Heterobimetallic Ru(ii)/Fe(ii) complexes as potent anticancer agents against breast cancer cells, inducing apoptosis through multiple targets.

Antimetastatic activity, high selectivity and cytotoxicity for human tumor cell lines make ruthenium(ii) complexes attractive for the development of new chemotherapeutic agents for cancer treatment. In this study, cytotoxic activities and the possible mechanism of cell death induced by three ruthenium complexes were evaluated, [Ru(MIm)(bipy)(dppf)]PF6 (1), [RuCl(Im)(bipy)(dppf)]PF6 (2) and [Ru(tzdt)(bipy)(dppf)]PF6 (3). The results showed high cytotoxicity and selectivity indexes for the human triple-negative breast tumor cell line (MDA-MB-231) with IC50 value and selectivity index for complex 1 (IC50 = 0.33 ± 0.03 µM, SI = 4.48), complex 2 (IC50 = 0.80 ± 0.06 µM, SI = 2.31) and complex 3 (IC50 = 0.48 ± 0.02 µM, SI = 3.87). The mechanism of cell death induced in MDA-MB-231 cells, after treatment with complexes 1-3, indicated apoptosis of the cells as a consequence of the increase in the percentage of cells in the Sub-G1 phase in the cell cycle analysis, characteristic morphological changes and the presence of apoptotic cells labeled with Annexin-V. Multiple targets of action were identified for complexes 1 and 3 with an induction of DNA damage in cells treated with complexes 1 and 3, mitochondrial depolarization with a reduction in mitochondrial membrane potential, an increase in reactive oxygen species levels and increased expression levels of caspase 3 and p53. In addition, antimetastatic activities for complexes 1 and 3 were observed by inhibition of cell migration by the wound healing assay and Boyden chamber assay, as well as inhibition of angiogenesis caused by MDA-MB-231 tumor cells in the CAM model.

Presence of antigenotoxic and anticytotoxic effects of the chalcone 1E,4E-1-(4-chlorophenyl)-5-(2,6,6-trimethylcyclohexen-1-yl)penta-1,4-dien-3-one using in vitro and in vivo assays.

Chalcones are chemically defined as α,ß-unsaturated ketones with a 1,3-diphenyl-2-propen-1-one nucleus. These compounds occur naturally in plants and are considered precursors of flavonoids. Given that evaluating genetic toxicology tests is essential in investigating the safe use and chemopreventive potential of different natural and synthetic compounds, this study aimed to assess the genotoxic, cytotoxic, antigenotoxic, and anticytotoxic activity of the chalcone 1E,4E-1-(4-chlorophenyl)-5-(2,6,6-trimethylcyclohexen-1-yl)penta-1,4-dien-3-one (CAB7ß). The CAB7ß was synthesized via Claisen-Schmidt reaction. The Ames test was applied using the co-treatment model as well as a micronucleus assay of mouse bone marrow with co-, pre- and post-treatment models. Our results indicate no genotoxic effect for CAB7ß in any of the tests applied. At all the concentrations used, CAB7ß showed a significant DNA protective effect against the mutagenic action of 4-nitroquinoline-1-oxide and sodium azide according to the Ames test, and against doxorubicin in the co-, pre- and post-treatment models of the micronucleus assay. CAB7ß alone displayed cytotoxic activity in the micronucleus test. At concentrations of 12,5 and 50 µg/plate, CAB7ß showed a moderate cytotoxic profile only in Salmonella typhimurium strain TA98. However, an anticytotoxic effect was observed against S. typhimurium strain TA100 for all the concentrations tested and during co-, pre- and post-treatment in the micronucleus assay. It was concluded that CAB7ß exhibited a slightly cytotoxic effect in S. typhimurium strain TA98 and significant antigenotoxic and anticytotoxic effects in cells of mouse, making it a promising candidate in chemoprevention and possibly in the development of new cancer treatments.

Assessment of the genotoxic and antigenotoxic activities of the aqueous solution of Caesalpinia ferrea (tul. ) Martius (Fabacae) fruit

Introduction: the inner bark of Caesalpinia ferrea (tul.) Martius (Fabacae), C. ferrea), popularly known as jucá, has been used in alternative medicine to treat wounds, bruises, asthma and chronic cough. Furthermore, the fruits of this species are used as antidiarrheals, decongestants and in healing, and their roots as antipyretics. Objective: to assess the possible genotoxic and antigenotoxic activities of the aqueous solution of the C. ferrea fruit. Methods: this study used the Ames test in Salmonella typhimurium strains and the micronucleus test in mouse bone marrow. Results: the Ames test results for the C. ferrea solution were not mutagenic in the Salmonella typhimurium TA100 strain in any of the doses tested. However, a protective effect against the action of sodium azide was shown in the TA100 strain at all the doses used. The micronucleus test indicated that the C. ferrea aqueous solution showed no mutagenic or antimutagenic effect. Conclusions: it was possible to conclude that the aqueous solution of the C. ferrea fruit showed no mutagenic effect in bacteria and mice, but an antimutagenic effect in bacteria.

Introducción: la corteza interna de Caesalpinia ferrea (tul.) Martius (Fabacae), C. ferrea, popularmente conocida como jucá, se ha utilizado en medicina alternativa para tratar heridas, hematomas, asma y tos crónica. Además, los frutos de esta especie se usan como antidiarreicos, descongestivos y en curación, y sus raíces como antipiréticos. Objetivo: evaluar las posibles actividades genotóxicas y antigenotóxicas de la solución acuosa del fruto de C. ferrea. Métodos: se utilizó la prueba de Ames en cepas de Salmonella typhimurium y la prueba de micronúcleo en médula ósea de ratón. Resultados: los resultados de la prueba de Ames para la solución de C. ferrea no fueron mutagénicos en la cepa TA100 de Salmonella typhimurium en ninguna de las dosis probadas. Sin embargo, se demostró un efecto protector contra la acción de la azida sódica en la cepa TA100 en todas las dosis utilizadas. La prueba de micronúcleos indicó que la solución acuosa de C. ferrea no mostró efecto mutagénico o antimutagénico. Conclusiones: la solución acuosa del fruto de C. ferrea no mostró efecto mutagénico en bacterias y ratones, sino un efecto antimutagénico en bacterias.

Antiangiogenic potential of Jatropha curcas latex in the chick chorioallantoic membrane model / Potencial antiangiogênico do látex de Jatropha curcas em modelo de membrana corioalantoica de embrião de galinha

AIMS: To perform a physicochemical and phytochemical characterization of Jatropha curcas latex and to investigate its antiangiogenic potential. METHODS: We performed an initial physicochemical characterization of J. curcas latex using thermal gravimetric analyses and Fourier Transform Infrared spectroscopy. After that, phenols, tannins and flavonoids were quantified. Finally, the potential of J. curcas latex to inhibit angiogenesis was evaluated using the chick chorioallantoic membrane model. Five groups of 20 fertilized chicken eggs each had the chorioallantoic membrane exposed to the following solutions: (1) water, negative control; (2) dexamethasone, angiogenesis inhibitor; (3) Regederm®, positive control; (4) 25% J. curcas latex diluted in water; (5) 50% J. curcas latex diluted in water; and (6) J. curcas crude latex. Analysis of the newly-formed vascular net was made through captured images and quantification of the number of pixels. Histological analyses were performed to evaluate the inflammation, neovascularization, and hyperemia parameters. The results were statically analyzed with a significance level set at p<0.05. RESULTS: Physicochemical characterization showed that J. curcas latex presented a low amount of cis-1.4-polyisoprene, which reduced its elasticity and thermal stability. Phytochemical analyses of J. curcas latex identified a substantial amount of phenols, tannins, and flavonoids (51.9%, 11.8%, and 0.07% respectively). Using a chick chorioallantoic membrane assay, we demonstrated the antiangiogenic potential of J. curcas latex. The latex induced a decrease in the vascularization of the membranes when compared with neutral and positive controls (water and Regederm®). However, when compared with the negative control (dexamethasone), higher J. curcas latex concentrations showed no significant differences. CONCLUSIONS: J. curcas latex showed low thermal stability, and consisted of phenols, tannins, and flavonoids, but little or no rubber. Moreover, this latex demonstrated a significant antiangiogenic activity on a chick chorioallantoic membrane model. The combination of antimutagenic, cytotoxic, antioxidant and antiangiogenic properties makes J. curcas latex a potential target for the development of new drugs.

OBJETIVOS: Realizar uma caracterização físico-química e fitoquímica do látex de Jatropha curcas e investigar o seu potencial antiangiogênico. MÉTODOS: foi realizada uma caracterização físico-química inicial do látex de J. curcas utilizando as análises termogravimétricas e a espectroscopia com a Transformada de Fourier. Depois disso, fenóis, taninos e flavonoides foram quantificados. Finalmente, o potencial do látex de J. curcas em inibir a angiogênese foi avaliado através do uso de modelo de membrana corioalantoica de embrião de galinha. Cinco grupos, cada um com 20 ovos de galinha fertilizados, tiveram a membrana corioalantoica exposta às seguintes soluções: (1) água, controle negativo; (2) dexametasona, inibidor da angiogênese; (3) Regederm®, controle positivo; (4) 25% de látex de J. curcas diluído em água; (5) 50% de látex de J. curcas diluído em água; e (6) látex bruto de J. curcas. A análise da rede vascular recém-formada foi feita por meio de imagens capturadas e quantificação do número de pixels. Análises histológicas foram realizadas para avaliar os parâmetros de inflamação, neovascularização e hiperemia. Os resultados foram analisados estaticamente com nível de significância estabelecido em p<0,05. RESULTADOS: A caracterização físico-química mostrou que o látex de J. curcas apresenta uma baixa quantidade de cis-1,4-poliisopreno, o que reduz sua elasticidade e estabilidade térmica. Análises fitoquímicas do látex de J. curcas identificaram uma quantidade significativa de fenóis, taninos e flavonoides (51,9%, 11,8% e 0,07% respectivamente). Usando o modelo de membrana corioalantoica de ovo de galinha embrionado, demonstrou-se o potencial antiangiogênico do látex de J. curcas. O látex induziu a diminuição da vascularização das membranas, em comparação aos grupos controle neutro e positivo (água e Regederm®). CONCLUSÕES: O látex de J. curcas apresentou baixa estabilidade térmica, ausência ou pouca quantidade de borracha e presença de fenóis, taninos e flavonoides em sua composição. Além disso, apresentou alta atividade antiangiogênica no modelo de membrana corioalantoica de embrião de galinha. A combinação de propriedades antimutagênicas, citotóxicas, anti-inflamatórias, antioxidantes e antiangiogênicas faz com que o látex de J. curcas seja um alvo potencial para o desenvolvimento de novos medicamentos.

Acute toxic effects of ruthenium (II)/amino acid/diphosphine complexes on Swiss mice and zebrafish embryos.

Anticancer potential of ruthenium complexes has been widely investigated, but safety evaluation studies are still scarce. Despite of ruthenium-based anticancer agents are known to cause fewer side effects compared to other metal-based drugs, these compounds are not fully free of toxicity, causing mainly nephrotoxicity. Based on the promising results from antitumor activity of the complexes [Ru(L-Met)(bipy)(dppb)]PF6 (RuMet) and [Ru(L-Trp)(bipy)(dppb)]PF6 (RuTrp), for the first time we investigated the toxicity profile of these complexes in rodent and zebrafish models. The acute oral toxicity was evaluated in Swiss mice. The mutagenic and genotoxic potential was determined by a combination of Micronucleus (MN) and Comet assay protocols, after exposure of Swiss mice to RuMet and RuTrp in therapeutic doses. Zebrafish embryos were exposed to these complexes, and their development observed up to 96 h post-fertilization. RuMet and RuTrp complexes showed low acute oral toxicity. Recorded behavioral changes were not recorded, nor were macroscopic morphological changes or structural modifications in the liver and kidneys. These complexes did not cause genetic toxicity, presenting a lack of micronuclei formation and low DNA damage induction in the cells from Swiss mice. In contradiction, cisplatin treatment exhibited high mutagenicity and genotoxicity. RuMet and RuTrp showed low toxicity in the embryo development of zebrafish. The RuMet and RuTrp complexes demonstrated low toxicity in the two study models, an interesting property in preclinical studies for novel anticancer agents.

Optical properties and antiangiogenic activity of a chalcone derivate.

Chalcones and their derivatives exhibit numerous pharmacological activities such as antibacterial, antifungal, cytotoxic, antinociceptive and anti-inflammatory. Recently, they have been assessed aiming for novel application in nonlinear optics and in the treatment of immune diseases and cancers. In this study, we investigate the optical properties of synthetic chalcona 1E,4E-1-(4-chlorophenyl)-5-(2,6,6-trimethylcyclohexen-1-yl)penta-1,4-dien-3-one (CAB7ß) and its antiangiogenic potential using the chorioallantoic membrane (CAM) with the S180 sarcoma cell line. Experimental and theoretical results show intense absorption in the UVA-UVC region, which is associated with a π → π* transition with intramolecular charge transfer from the trimethyl-cyclohexen-1-yl ring to the chlorophenyl ring. Quantum chemical calculations of the first hyperpolarizability, accounting for both solvent and frequency dispersion effects, are in very good concordance with hyper-Rayleigh scattering measurements. In addition, two-photon absorption allowed band centered at 650 nm was observed. Concerning antiangiogenic activity, CAB7ß causes a significant reduction in the total number, junctions, length and caliber of blood vessels stimulated by S180 cells reducing the presence of blood vessels, inflammatory cells and others elements related to angiogenic process. It is found that CAB7ß is a versatile compound and a promising candidate for linear and nonlinear optical applications, in therapy against sarcoma and phototherapy.

IMMUNOMODULATING EFFECTS OF THE PURIFIED HEV B 13 FRACTION ON SEPTIC RATS.

BACKGROUND: Sepsis is a potentially life-threatening complication of an infection that occurs when chemicals released into the bloodstream to fight the infection trigger inflammatory responses throughout the body, especially in the acute phase of the disease, producing excessive pro-inflammatory cytokines, leading to multiple organ injury and death. The Hev b 13 fraction has demonstrated biological activity capable of inducing IL-10 production and shrinking inflammatory disease lesions. AIM: To investigate the immunomodulating effects of the Hev b 13 fraction on septic rats. METHODS: Acinetobacter baumannii was injected into the peritoneal cavity of the animals after sustaining a lesion in the pancreas, with the stomach as an entry point. After 10 h of infection, they were euthanized for blood and lung collection, followed by total and differential leukocyte count, determination of cytokine level and histopathological analysis. RESULTS: Administering a single dose of the Hev b 13 fraction 2 h after sepsis induction significantly decreased total leukocyte count. Higher IL-10 and IL-4 and lower IL-6 production shrank the lung tissue lesions compared to the control groups. CONCLUSION: The Hev b 13 fraction exhibits an anti-inflammatory tendency, with potential for sepsis treatment.

Antitumor effectiveness and mechanism of action of Ru(II)/amino acid/diphosphine complexes in the peritoneal carcinomatosis progression.

Peritoneal carcinomatosis is considered as a potentially lethal clinical condition, and the therapeutic options are limited. The antitumor effectiveness of the [Ru(l-Met)(bipy)(dppb)]PF6(1) and the [Ru(l-Trp)(bipy)(dppb)]PF6(2) complexes were evaluated in the peritoneal carcinomatosis model, Ehrlich ascites carcinoma-bearing Swiss mice. This is the first study that evaluated the effect of Ru(II)/amino acid complexes for antitumor activity in vivo. Complexes 1 and 2 (2 and 6 mg kg-1) showed tumor growth inhibition ranging from moderate to high. The mean survival time of animal groups treated with complexes 1 and 2 was higher than in the negative and vehicle control groups. The induction of Ehrlich ascites carcinoma in mice led to alterations in hematological and biochemical parameters, and not the treatment with complexes 1 and 2. The treatment of Ehrlich ascites carcinoma-bearing mice with complexes 1 and 2 increased the number of Annexin V positive cells and cleaved caspase-3 levels and induced changes in the cell morphology and in the cell cycle phases by induction of sub-G1 and G0/G1 cell cycle arrest. In addition, these complexes reduce angiogenesis induced by Ehrlich ascites carcinoma cells in chick embryo chorioallantoic membrane model. The treatment with the LAT1 inhibitor decreased the sensitivity of the Ehrlich ascites carcinoma cells to complexes 1 and 2 in vitro-which suggests that the LAT1 could be related to the mechanism of action of amino acid/ruthenium(II) complexes, consequently decreasing the glucose uptake. Therefore, these complexes could be used to reduce tumor growth and increase mean survival time with less toxicity than cisplatin. Besides, these complexes induce apoptosis by combination of different mechanism of action.

cis-[RuCl(BzCN)(bipy)(dppe)]PF6 induces anti-angiogenesis and apoptosis by a mechanism of caspase-dependent involving DNA damage, PARP activation, and Tp53 induction in Ehrlich tumor cells.

Antimetastatic activities, low toxicity to normal cells and high selectivity for tumor cells make of the ruthenium complexes promising candidates in the search for develop new chemotherapeutic agents for the treatment of cancer. This study aimed to determine the cytotoxic, genotoxic and to elucidate the signaling pathway involved in the death cell process induced by cis-[RuCl(BzCN)(bipy)(dppb)]PF6(1) and cis-[RuCl(BzCN)(bipy)(dppe)]PF6(2) in Ehrlich ascites carcinoma (EAC) in vitro. Moreover, we report for the first time the anti-angiogenic potential on chick embryo chorioallantoic membrane (CAM) model. Peripheral blood mononuclear cells (PBMC) were isolated from healthy controls with an age range of 20-30 years and used to calculate the selectivity index (SI). The complex 2 (IC50 = 8.5 ± 0.4/SI = 6.3) showed high cytotoxic and selectivity index against EAC cells than complex 1 (IC50 = 14.9 ± 0.2/SI = 0.2) using the MTT assay. Complex 2 induced DNA damage on Ehrlich tumor cells at concentrations and time periods evalueted. In consequence, it was observed an increase of Tp53 gene expression, G0/G1-arrest cells, and increased levels of cleaved PARP protein. Beside that, the treatment of EAC with complex 2 led to an increase in Annexin V-positive cells and apoptosis induction by Caspase-7. Additionally, the complex 2 inhibited the angiogenesis caused by Ehrlich tumor cells in CAM model. This complex is active and selective for Ehrlich tumor cells, inducing DNA damage, cell cycle arrest and cell death by caspase-dependent apoptosis involving PARP activation (PARP1), and Tp53 induction.

Antigenotoxicity protection of Carapa guianensis oil against mitomycin C and cyclophosphamide in mouse bone marrow.

The aim of this study was to evaluate the possible protective of C. guianensis oil against MMC and CP, which are direct- and indirect-acting chemical mutagens, using the micronucleus test. Three experiments were performed. First the C. guianensis oil was co-administered to mice at doses of 250, 500 and 1000 mg/kg bw with 4 mg/kg bw MMC or 50 mg/kg bw CP. Second, the mutagenic drug (CP) was administered ip 50 mg/kg bw and after 6 and 12 hours 250 and 500 mg/kg bw of C. guianensis oil were administered. In the last, C. guianensis oil was administrated (250 and 500 mg/kg bw) during five days and after it was administered ip 50 mg/kg bw CP. The results obtained showed that the C. guianensis oil is not cytotoxic neither genotoxic to mouse bone marrow. Regarding the antimutagenic effect, all doses of C. guianensis oil were significantly (p < 0.05) effective in reducing the frequency of micronucleated polychromatic erythrocytes, when compared with MMC or CP alone. Based on these results, our results suggest that the C. guianensis oil shows medicinal potential as an antimutagenic agent, modulating the mutagenicity caused by both direct- and indirect-acting chemical mutagens, in a mammalian model.

Immunomodulating effects of the purified hev b 13 fraction on septic rats / Efeitos imunomoduladores da fração purificada hev b 13 em ratos com sepse

ABSTRACT Background: Sepsis is a potentially life-threatening complication of an infection that occurs when chemicals released into the bloodstream to fight the infection trigger inflammatory responses throughout the body, especially in the acute phase of the disease, producing excessive pro-inflammatory cytokines, leading to multiple organ injury and death. The Hev b 13 fraction has demonstrated biological activity capable of inducing IL-10 production and shrinking inflammatory disease lesions. Aim: To investigate the immunomodulating effects of the Hev b 13 fraction on septic rats. Methods: Acinetobacter baumannii was injected into the peritoneal cavity of the animals after sustaining a lesion in the pancreas, with the stomach as an entry point. After 10 h of infection, they were euthanized for blood and lung collection, followed by total and differential leukocyte count, determination of cytokine level and histopathological analysis. Results: Administering a single dose of the Hev b 13 fraction 2 h after sepsis induction significantly decreased total leukocyte count. Higher IL-10 and IL-4 and lower IL-6 production shrank the lung tissue lesions compared to the control groups. Conclusion: The Hev b 13 fraction exhibits an anti-inflammatory tendency, with potential for sepsis treatment.

RESUMO Racional: Sepse se correlaciona com a ruptura do complexo equilíbrio entre os mediadores inflamatórios, que principalmente na fase aguda da doença, produz exacerbadamente citocinas pró-inflamatórias levando a lesão de múltiplos órgãos e morte. A fração Hev b 13 tem demonstrado atividade biológica capaz de induzir a produção de IL-10 e regredir lesões de doenças inflamatórias. Objetivo: Investigar os efeitos imunomoduladores da fração Hev b 13 em ratos com sepse. Métodos: Foi injetado Acinetobacter baumannii na cavidade peritoneal dos animais após lesão no pâncreas e estômago como porta de entrada. Após 10 h de infecção, foi realizada eutanásia para coleta de sangue e pulmões, em seguida, contagem total e diferencial de leucócitos, dosagem de citocinas e histopatologia para análise. Resultados: A administração de dose única da fração Hev b 13, 2 h após a indução de sepse, diminuiu significativamente a contagem total de leucócitos. Associado a maior produção de IL-10 e IL-4, e menor de IL-6, atenuou as lesões nos tecidos pulmonares em comparação com os grupos controles. Conclusão: A fração Hev b 13 apresenta tendência anti-inflamatória, com potencialidades no tratamento da sepse.

Antigenotoxicity protection of Carapa guianensis oil against mitomycin C and cyclophosphamide in mouse bone marrow

ABSTRACT The aim of this study was to evaluate the possible protective of C. guianensis oil against MMC and CP, which are direct- and indirect-acting chemical mutagens, using the micronucleus test. Three experiments were performed. First the C. guianensis oil was co-administered to mice at doses of 250, 500 and 1000 mg/kg bw with 4 mg/kg bw MMC or 50 mg/kg bw CP. Second, the mutagenic drug (CP) was administered ip 50 mg/kg bw and after 6 and 12 hours 250 and 500 mg/kg bw of C. guianensis oil were administered. In the last, C. guianensis oil was administrated (250 and 500 mg/kg bw) during five days and after it was administered ip 50 mg/kg bw CP. The results obtained showed that the C. guianensis oil is not cytotoxic neither genotoxic to mouse bone marrow. Regarding the antimutagenic effect, all doses of C. guianensis oil were significantly (p < 0.05) effective in reducing the frequency of micronucleated polychromatic erythrocytes, when compared with MMC or CP alone. Based on these results, our results suggest that the C. guianensis oil shows medicinal potential as an antimutagenic agent, modulating the mutagenicity caused by both direct- and indirect-acting chemical mutagens, in a mammalian model.

Physicochemical/photophysical characterization and angiogenic properties of Curcuma longa essential oil.

This study analyzed the physicochemical and photophysical properties of essential oil of Curcuma longa and its angiogenic potential. The results showed that curcumin is the main fluorescent component present in the oil, although the amount is relatively small. The experimental chorioallantoic membrane model was used to evaluate angiogenic activity, showing a significant increase in the vascular network of Curcuma longa and positive control groups when compared to the neutral and inhibitor controls (P <0.05), but no significant difference was found between Curcuma longa essential oil and the positive control (P >0.05). Histological analysis showed extensive neovascularization, hyperemia and inflammation in the positive control group and Curcuma longa when compared to other controls (P <0.05), characteristic factors of the angiogenesis process. In conclusion, Curcuma longa oil showed considerable proangiogenic activity and could be a potential compound in medical applications.

Chemopreventive effect and angiogenic activity of punicalagin isolated from leaves of Lafoensia pacari A. St.-Hil.

Punicalagin is the major ellagitannin constituent from leaves of Lafoensia pacari, a Brazilian medicinal plant widely used for the treatment of peptic ulcer and wound healing. Genotoxic, cytotoxic, antigenotoxic, and anticytotoxic effects of punicalagin were assessed using micronucleus (MN) test and comet assay in mice. Due to the extensive use of L. pacari in the wound healing process, we also assessed the angiogenic activity of punicalagin using the chick chorioallantoic membrane (CAM) angiogenic assay. The highest dose of punicalagin (50mg/kg) showed significant cytotoxic effect by MN test and in the co-treatment with cyclophosphamide (CPA), this cytotoxicity was enhanced. Co-treatment, pre-treatment and post-treatment of punicalagin with CPA led to a significant reduction in the number of DNA breaks and in the frequency of CPA-induced MN, indicating antigenotoxic effect. Using the CAM model, punicalagin exhibited angiogenic activity in all doses mainly at the lowest concentration (12.5µg/µL). Therefore, these findings indicate an effective chemopreventive role of punicalagin and a high capacity to induce DNA repair. Also, the angiogenic activity presented by punicalagin in this study could contribute for the processes of tissue repairing and wound healing.

EFFECTS OF TOPICAL TREATMENT WITH EUPHORBIA TIRUCALLI LATEX ON THE SURVIVAL AND INTESTINAL ADHESIONS IN RATS WITH EXPERIMENTAL PERITONITIS / EFEITOS DO TRATAMENTO TÓPICO COM O LÁTEX DA EUPHORBIA TIRUCALLI NA SOBREVIDA E NAS ADERÊNCIAS INTESTINAIS DE RATOS COM PERITONITE EXPERIMENTAL

Background: The use of plants of the family Euphorbiaceae, particularly Euphorbia tirucalli (avelós) has been popularly widespread for treating a variety of diseases of infectious, tumoral, and inflammatory. Aim: To demonstrated antimicrobial and immunomodulatory effects of these extracts, evaluating the effect of a topical treatment with an aqueous solution of avelós latex on the survival and on intestinal adhesions in rats with experimental peritonitis. Methods: Peritonitis was induced in 24 Wistar rats, that were randomized into four groups of six as follows: (1) Control group (n=6), no treatment; (2) Antibiotic group (n=6), treatment with a single intramuscular dose of antibiotic Unasyn; (3) Saline group (n=6), the abdominal cavity was washed with 0.9% saline; and (4) E.tirucalli group (n=6), the abdominal cavity was washed with E. tirucalli at a concentration of 12 mg/ml. The animals that died were necropsied, and the time of death was recorded. The survivors were killed on postoperative day 11, and necropsy was subsequently performed for evaluation of the intestinal adhesions. Results: Significant differences were observed in the control and antibiotic groups (p<0.01) with respect to the survival hours when compared with the saline and E. tirucalli groups. There was no significant difference (p>0.05) in the survival of animals in the saline andE. tirucalli groups; however, one animal died in the saline group. Necropsy of the animals in the saline and E. tirucalligroups showed strong adhesions resistant to manipulation, between the intestinal loops and abdominal wall. The remaining groups did not show any adhesions. Conclusions: Topical treatment with E. tirucalli latex stimulated an increased formation of intestinal adhesions and prevented the death of all animals with peritonitis.

Racional: O uso de plantas da família Euphorbiaceae, principalmente a Euphorbia tirucalli (avelós), tem sido popularmente difundido para o tratamento de uma variedade de doenças de natureza infecciosa, tumoral e inflamatória. Objetivo: Avaliar o efeito do tratamento tópico com a solução aquosa do látex do avelós na sobrevida e nas aderências intestinas de ratos com peritonite experimental. Métodos: Foi induzido peritonite em 24 ratos Wistar e randomizados em quatro grupos de seis, assim distribuídos: 1) Controle - (n=6), nenhum tratamento; 2) Antibiótico - (n=6), tratamento com dose única intramuscular de antibiótico Unasyn (Pfizer - São Paulo); 3) Salina - (n=6), lavagem da cavidade abdominal com solução fisiológica 0,9%; 4) E.Tirucalli - (n=6), lavagem da cavidade abdominal com E. tirucalli na concentração de 12 mg/ml. Os animais que morreram foram submetidos à necropsia e o horário do óbito anotado. Os sobreviventes foram submetidos à eutanásia no 11odia de pós-operatório e, posteriormente, realizou-se a necropsia para avaliação da formação de aderências. Resultados: Os grupos controle e antibiótico obtiveram diferença significativa (p<0,01) com relação às horas de vida entre os grupos salina e E. tirucalli. Não houve diferença significativa (p>0,05) na sobrevida dos animais dos grupos salina e E. tirucalli, no entanto, houve um óbito no grupo salina. A necropsia dos animais dos grupos salina e E. tirucalli mostrou aderências firmes e resistentes à manipulação entre alças intestinais e parede abdominal. Os demais grupos não tiveram formação de aderências. Conclusão: O tratamento tópico com o látex da E. tirucalli estimulou maior formação de aderências intestinais e evitou o óbito de todos animais com peritonite até o período avaliado.

Angiogenic activity of sucupira (Pterodon emarginatus) oil / Atividade angiogênica do óleo de sucupira (Pterodon emarginatus)

Aims: To assess the angiogenic activity of sucupira (Pterodon emarginatus) oil, popularly used as an antirheumatic, analgesic, antimicrobial, and anti-inflammatory agent. Methods: The chick (Gallus domesticus) embryo chorioallantoic membrane (CAM) assay was used as an experimental model, incubated in a temperature- and humidity- controlled automatic incubator during the first five days. A circular hole was made at the wider base of the eggshell on the fifth day. After 13 days of incubation, filter paper discs containing sucupira oil and control substances were inserted into the CAM. Subsequently, the CAMs were removed and submitted to analysis and quantification of the vascular network. The nonparametric KruskalWallis test, followed by Dunn's test, was used for the statistical analysis, and differences between the samples were set at a 5% level (p<0.05). Results: Sucupira oil at a concentration of 1g/mL induced a significant increase in the percentage area of the CAM vascular network of the embryonated chicken eggs, when compared with the neutral control (water) and inhibitory control (dexamethasone) groups (p<0.001). However, no significant difference in the induction of the CAM vascular network (p>0.05) was demonstrated between the oil and the positive control (Biocure) groups. Conclusions: The sucupira (Pterodon emarginatus) bean oil induced the formation of new blood vessels under the experimental conditions of the present study.

Objetivos: Avaliar a atividade angiogênica do óleo de sucupira (Pterodon emarginatus), popularmente utilizado como antirreumático, analgésico, antimicrobiano e anti-inflamatório. Métodos: Foi aplicado como modelo experimental o ensaio na membrana corioalantóidea (MCA) de ovo embrionado de galinha (Gallus domesticus). Os ovos foram incubados em estufa automática com temperatura e umidade controladas durante os primeiros cinco dias. No quinto dia foi realizada uma abertura circular na base maior da casca do ovo. No 13º dia de incubação, discos de papel de filtro contendo óleo de sucupira e as substâncias controle foram inseridos sobre vasos da MCA. Posteriormente, as MCAs foram removidas e passaram por análise e quantificação da rede vascular. Para análise estatística foi utilizado o teste não paramétrico de Kruskal-Wallis, seguido de Dunn, sendo os valores aceitáveis para diferenças significativas entre as amostras de p<0.05. Resultados: O óleo da fava de sucupira na concentração de 1g/mL induziu um aumento significativo na área de porcentagem da rede vascular na MCA do ovo embrionado de galinha, quando comparados aos grupos controles neutro (água) e inibidor (dexametasona) (p<0.001). Porém, não foi demonstrada diferença significativa entre o óleo e o controle positivo (Biocure) na indução da rede vascular na MCA (p>0.05). Conclusões: O óleo da fava de sucupira (Pterodon emarginatus), nas condições deste experimento, foi responsável pela formação de novos vasos sanguíneos.

EFFECTS OF TOPICAL TREATMENT WITH EUPHORBIA TIRUCALLI LATEX ON THE SURVIVAL AND INTESTINAL ADHESIONS IN RATS WITH EXPERIMENTAL PERITONITIS.

BACKGROUND: The use of plants of the family Euphorbiaceae, particularly Euphorbia tirucalli (avelós) has been popularly widespread for treating a variety of diseases of infectious, tumoral, and inflammatory. AIM: To demonstrated antimicrobial and immunomodulatory effects of these extracts, evaluating the effect of a topical treatment with an aqueous solution of avelós latex on the survival and on intestinal adhesions in rats with experimental peritonitis. METHODS: Peritonitis was induced in 24 Wistar rats, that were randomized into four groups of six as follows: (1) Control group (n=6), no treatment; (2) Antibiotic group (n=6), treatment with a single intramuscular dose of antibiotic Unasyn; (3) Saline group (n=6), the abdominal cavity was washed with 0.9% saline; and (4) E.tirucalli group (n=6), the abdominal cavity was washed with E. tirucalli at a concentration of 12 mg/ml. The animals that died were necropsied, and the time of death was recorded. The survivors were killed on postoperative day 11, and necropsy was subsequently performed for evaluation of the intestinal adhesions. RESULTS: Significant differences were observed in the control and antibiotic groups (p<0.01) with respect to the survival hours when compared with the saline and E. tirucalli groups. There was no significant difference (p>0.05) in the survival of animals in the saline andE. tirucalli groups; however, one animal died in the saline group. Necropsy of the animals in the saline and E. tirucalligroups showed strong adhesions resistant to manipulation, between the intestinal loops and abdominal wall. The remaining groups did not show any adhesions. CONCLUSIONS: Topical treatment with E. tirucalli latex stimulated an increased formation of intestinal adhesions and prevented the death of all animals with peritonitis.