Hepatoprotective effect of Phytosome Curcumin against paracetamol-induced liver toxicity in mice

Abstract Curcuma longa, which contains curcumin as a major constituent, has been shown many pharmacological effects, but it is limited using in clinical due to low bioavailability. In this study, we developed a phytosome curcumin formulation and evaluated the hepatoprotective effect of phytosome curcumin on paracetamol induced liver damage in mice. Phytosome curcumin (equivalent to curcumin 100 and 200 mg/kg body weight) and curcumin (200 mg/kg body weight) were given by gastrically and toxicity was induced by paracetamol (500 mg/kg) during 7 days. On the final day animals were sacrificed and liver function markers (ALT, AST), hepatic antioxidants (SOD, CAT and GPx) and lipid peroxidation in liver homogenate were estimated. Our data showed that phytosome has stronger hepatoprotective effect compared to curcumin-free. Administration of phytosome curcumin effectively suppressed paracetamol-induced liver injury evidenced by a reduction of lipid peroxidation level, and elevated enzymatic antioxidant activities of superoxide dismutase, catalase, glutathione peroxidase in mice liver tissue. Our study suggests that phytosome curcumin has strong antioxidant activity and potential hepatoprotective effects.

Hydroethanolic Allium sativum extract accelerates excision wound healing: evidence for roles of mast-cell infiltration and intracytoplasmic carbohydrate ratio

ABSTRACT The present study was designed to evaluate the in vivo effect of Allium sativum (garlic) hydroalcoholic extract on wound healing in rats. For this purpose, 72 mature Wistar rats were divided into four groups (n=18/each) to receive no treatment, placebo, Cicalfate(r), or 2% Allium sativum (AS) extract, administered topically to the wound area, for 21 days. Following the experimental period, tissue samples were dissected out and underwent to histopathological analyses. Fibroblasts, fibrocytes, mast cells, intra-cytoplasmic carbohydrate ratio, neovascularization, collagen deposition, and re-epithelialization were analyzed in all groups. Animals in the treated groups showed significant enhancement in fibroblast, fibrocyte, and mast-cell distribution. Significantly higher neovascularization was observed on day 3 after wound induction in AS-treated animals versus those in the placebo, Cicalfate, and untreated groups (P<0.05). A dose-dependent, significantly higher intra-cytoplasmic carbohydrate storage was observed in treated animals. Our data show that AS promotes wound healing due to its preliminary impact on mast-cell distribution, which enhanced collagen synthesis and upregulated angiogenesis, and shortened the healing process by enhancing the intra-cytoplasmic carbohydrate ratio.

Changes in weight and body fat after use of tetracycline and Lactobacillus gasseri in rats

ABSTRACT Recent studies have shown a role of intestinal microbiota in obesity. The consumption of antibiotics in the last 70 years has led to changes in intestinal microbiota, which has led to weight gain and body fat accumulation. To evaluate the possibility of weight gain induced by antibiotics and the possible protective effect of probiotics, we divided 45 animals (Rattus norvegicus) into groups and administered the following treatments over two weeks: tetracycline, tetracycline + Lactobacillus gasseri, and NaCl. The animals were weighed over the course of 8 weeks, and at the end of the treatment period, they were measured and subjected to bioelectrical impedance analysis. The results show that the group receiving tetracycline alone had a higher body mass index (p=0.030), a greater Lee index (p=0.008), and a lower body water percentage than the control group, indicating a greater accumulation of body fat. The group receiving the probiotics with tetracycline presented similar results to the control group, indicating a possible protective effect of body fat accumulation in the group receiving tetracycline alone. The results show that tetracycline increased the concentration of body fat, and the use of probiotics was associated with an ability to protect the animals from the pro-obesity effect.

Compostos fenólicos da casca de Handroanthus heptaphyllus (Mart. ) Mattos e efeitos do extrato aquoso no perfil lipídico, glicêmico e na lipoperoxidação em ratos diabéticos / Phenolic compounds from Handroanthus heptaphyllus (Mart. ) Mattos bark and effects of aqueous extract on lipid profile, glucose, and lipid peroxidation in diabetic rats

RESUMO A preocupação com o tratamento do Diabetes mellitus (DM) leva a uma crescente busca por terapias alternativas, como o uso de plantas medicinais, entre as quais, destaca-se o uso de Handroanthus heptaphyllus (Mart.) Mattos (popular Ipê roxo). Neste estudo realizamos a investigação química da presença de compostos fenólicos em H. heptaphyllus e o efeito do tratamento com o extrato aquoso da casca desta planta em parâmetros bioquímicos e nos níveis de lipoperoxidação tecidual e plasmática em animais diabéticos. Metodologia: Ratos Wistar machos foram submetidos ao desenvolvimento do quadro de DM por meio da administração intraperitoneal (IP) de Aloxano monohidrato (150 mg/Kg IP). Após a confirmação de hiperglicemia (>200 mg dL-1), os animais foram distribuídos nos grupos Diabético (D; n=6) e Diabético Tratado (DT; n=6). O tratamento consistiu na administração diária do extrato aquoso da casca de H. heptaphyllus via oral (v.o.) (150mg/Kg v.o.) por quatro semanas. O extrato aquoso foi analisado qualitativamente por cromatografia de camada delgada. Resultados: A análise qualitativa do extrato aquoso da casca indicou a presença de compostos fenólicos da subclasse flavonoides. O tratamento com o extrato aquoso reduziu a glicemia de jejum a partir da 3ª semana de tratamento, melhorou a resposta glicêmica à sobrecarga de glicose, diminuiu os níveis de triglicerídeos e índice LDL (Triglicerídeos/HDL). Estes resultados sugerem o uso terapêutico do extrato aquoso das cascas de H. heptaphyllus no tratamento do DM.

ABSTRACT Alternative medicine for diabetes mellitus (DM) treatment represents a growing research area on the use of medicinal plants, of which Handroanthus heptaphyllus (mart.) Mattos (popularly known as purple ipe) is most prominent. In this study, we investigated the presence of phenolic compounds and the effects of treatment with aqueous extract of in H. heptaphyllus in biochemical profile in plasma and the levels of lipid peroxidation in tissues and plasma in diabetic animals. Male Wistar rats were induced to develop DM through intraperitoneal (IP) administration of alloxan monohydrate (150 mg/kg IP). Once hyperglycemia (>200 mg dL-1) was confirmed, the animals were divided into the Diabetic (D; n=6) and Treated Diabetic (TD; n=6) groups. The TD group received daily administration (150 mg/kg v.o.) of aqueous extract of H. heptaphyllus for four weeks. The aqueous extract was also analyzed qualitatively by layer chromatography. Qualitative analysis of the aqueous extract of the bark indicated the presence of phenolic compounds from the flavonoid subclass. The treatment with the aqueous extract reduced fasting blood glucose levels from the third week of treatment on, improved the glycemic response to the glucose tolerance test, and lowered the levels of triglycerides and the LDL index (triglycerides/HDL). These findings suggest therapeutic use of the aqueous extract of H. heptaphyllus bark in treating DM.

Efeitos citotóxicos, genotóxicos e epigenéticos do Bisfenol A em células HL-60, MCF-7 e em ratos / Cytotoxic, genotoxic and epigenetics effects of Bisphenol A on HL-60, MCF-7 cells and rats

Bisfenol A (BPA) é um insumo largamente utilizado na produção de plástico policarbonato e amplamente difundido no meio ambiente, levando o ser humano à exposição crônica desde o período intrauterino. A literatura aponta a possibilidade de BPA aumentar o risco de diversos tipos de câncer, mas são necessários estudos que possibilitem o entendimento de mecanismos pelos quais isso pode ocorrer. Neste trabalho foram investigados os efeitos do BPA ou nitro-BPA em células HL-60, MCF-7 e tecidos de ratos. Células HL-60 foram expostas ao BPA ou nitro-BPA nas concentrações de 25, 100 e 250 µM (0,1 % DMSO v/v) por 2, 24 ou 48 horas na presença ou ausência de H2O2 (40 nmol/5 x 104 células). Células MCF-7 foram expostas da mesma forma, sem o uso de H2O2, mas na presença e ausência de agonista (PCB) de receptor Ah. Ratos Sprague-Dawley machos receberam BPA diariamente ao longo de 4 semanas (50 mg/kg de peso corpóreo) por gavagem, na vigência e ausência de diabetes, com subsequente coleta de urina, fígado, rins, medula óssea e sangue. Nos experimentos com as células, a viabilidade, ciclo celular, fragmentação do DNA e a produção intracelular de espécies reativas de oxigênio (ROs) foram avaliadas por citometria de fluxo, a atividade da cadeia respiratória mitocondrial pelo ensaio do XTT, e a atividade de MPO de células HL-60 por ensaio de fluorescência, bem como a produção de •NO. A metilação e hidroximetilação global do DNA e os adutos 8-oxodG, CEdG, 1,N6-εdA, 1,N2-εdG e BPA-Gua no DNA das células, tecidos, meio de cultura e urina foram analisados por HPLC-ESI-MS/MS. O hemograma e mielograma dos animais foram obtidos no Laboratório de Hematologia Experimental da FCF USP. Observou-se que tanto BPA quanto nitro-BPA induziram a geração de ROS em células HL-60 logo após 2h de incubação. BPA levou subsequentemente à perda de atividade da cadeia respiratória mitocondrial, aumento da permeabilidade da membrana plasmática, fragmentação do DNA, parada na fase G2/M do ciclo celular e hipermetilação acompanhada de hipohidroximetilação global do DNA. A citotoxicidade induzida pelas mesmas concentrações de nitro-BPA em células HL-60 foi menos pronunciada, sem perda de atividade da cadeia respiratória mitocondrial, com pouca fragmentação do DNA, mas com parada na fase G0/G1 do ciclo celular e indução de hipohidroximetilação global do DNA na presença de H2O2. Não foi observada a indução de adutos de DNA nas células HL-60 incubadas com BPA, mas sim de CEdG nas células incubadas com nitro-BPA. Os dados obtidos a partir da exposição das células HL-60 a BPA e nitro-BPA nos indicam que as duas moléculas provocam alterações metabólicas distintas nesse tipo celular, independentes da via estrogênica, que levam a alterações predominantemente epigenéticas (BPA) ou genéticas e epigenéticas (nitro-BPA), que podem ter consequências fenotípicas, como progressão maligna, que precisam ser investigadas. Foi observado que as células MCF-7 são mais resistentes que as células HL-60 à citotoxicidade induzida por BPA e nitro-BPA. Como resultado da exposição das células MCF-7 a BPA, houve pequeno aumento da permeabilidade da membrana plasmática (250 µM), indução dos níveis de ROS após 24 h (25 µM) e aumento da população de células em sub G1, ou seja, com DNA fragmentado (100 µM e 250 µM), mas sem alteração do ciclo celular. No caso de nitro-BPA, foi observada parada do ciclo celular em G2/M (25 µM, 100 µM e 250 µM), assim como aumento de permeabilidade da membrana plasmática após 24 h de incubação (25 µM, 250 µM), sem indução de ROS ou aumento de células em sub G1. Entretanto, observou-se aumento dos níveis de CEdG e 8-oxodG no DNA das células incubadas com BPA (100 µM, 250 µM) sem a ativação prévia de receptores Ah. A ativação dos receptores Ah com PCB levou a menor aumento do nível das lesões após as incubações com BPA. A maior resistência das células MCF-7 aos efeitos citotóxicos do BPA está provavelmente relacionada à ação estrogênica desse xenobiótico. A sinalização estrogênica juntamente com o aumento dos níveis de lesões no DNA aumenta a chance de mutações e de transformação maligna. Nas células com ativação do receptor Ah, BPA levou ainda ao aumento da hidroximetilação global, sem alteração da metilação global do DNA. Os animais não diabéticos expostos ao BPA apresentaram quantidades diminuídas de promielócitos, blastos e bastonetes na medula óssea (aplasia medular), sem alteração no hemograma. Houve aumento dos níveis de CEdG no fígado, da metilação e hidroximetilação global do DNA hepático, e não foi observada alteração das marcas epigenéticas e adutos de DNA no rim ou na urina. Os animais diabéticos expostos ao BPA apresentaram aumento do número de eosinófilos e linfócitos na medula óssea, podendo-se sugerir a indução de um estado inflamatório alérgico, e aumento do número total de hemácias circulantes e do hematócrito. Houve aumento dos níveis de CEdG, da metilação e hidroximetilação global do DNA hepático, aumento dos níveis de 8-oxodG no DNA renal, sem alteração das marcas epigenéticas no rim, e não foi observada alteração dos adutos de DNA na urina. Os dados obtidos apontam para a geração de ROS como uma importante via de cito- e genotoxicidade induzidas por BPA. Sua biotransformação para BPA-3,4- quinona nos modelos utilizados parece ter menor importância para os efeitos, uma vez que não foi detectada a lesão BPA-Gua em nenhuma amostra de DNA, meio de cultura das células ou urina dos animais. Alterações metabólicas induzidas por BPA e ROS podem favorecer as alterações das marcas epigenéticas observadas no DNA das células HL-60, MCF-7 e fígado dos animais. Todas essas alterações podem contribuir para a transformação maligna de células expostas ao BPA

Bisphenol A (BPA) is a compound widely used in polycarbonate plastic production and widespread in the environment, humans are chronic exposed to BPA in intrauterine period and entire life. The literature suggests the possibility of BPA increase the risk of developing cancers, but studies are required to enable the understanding of mechanisms by which this can occur. HL -60 cells were exposed to BPA or nitro-BPA at concentrations of 25, 100 and 250 uM (0.1% DMSO v/v) for 2, 24 or 48 hours in presence or absence of H2O2 (40 nmol/5x104 cells), MCF-7 cells followed a similar profile of exposure without the use of H2O2, but in presence or absence of Ah agonist receptor (PCB126). Male Sprague-Dawley rats received BPA daily over 4 weeks (50 mg/kg body weight) by gavage in presence and absence of diabetes, with subsequent collection of urine, liver, kidney, bone marrow and circulating blood. The viability, cell cycle, DNA fragmentation and the intracellular production of reactive oxygen species (ROS) was evaluated by flow cytometry, MPO activity and NO production was evaluated by fluorescence assay for HL- 60 cells, mitochondrial activity by XTT assay, and the global DNA methylation was checked by HPLC-PDA. DNA adducts 8-oxodG, CEdG, 1,N6-εdA, 1,N6-εdG and BPA-Gua were quantified by HPLC- ESI-MS/MS in DNA of cells, culture medium, urine and tissue collected from Sprague-Dawley rats. Blood count and bone marrow examination were obtained in collaboration with Experimental Hematology Laboratory of University of Sao Paulo We observed that both BPA and BPANO2 induced ROS generation in HL- 60 cells after 2 hours of incubation. BPA subsequently led to failure of mitochondrial respiratory chain activity, increased permeability of the plasma membrane, DNA fragmentation, arrest in G2/M phase of cell cycle, DNA hypermethylation with global hipohydroxymethylation. We saw low cytotoxicity in HL-60 cells induced by nitro-bpa n the same concentration, without loss of mitochondrial respiratory chain activity, discrete DNA fragmentation, but leading cell cycle to stopping at G0/G1 phase, and induction of DNA global hypermethylation. No lesions were observed in the DNA of HL-60 cells.The results obtained from the exposure of HL-60 cells to BPA and nitro-BPA indicate that these two molecules induce different metabolic abnormalities in this cell line, independent of estrogen pathway, leading to changes in epigenetic (BPA) or genetic and epigenetic (nitro-BPA) profile, that can induce phenotypic consequences such as malignant progression. It was observed along the study that MCF-7 cells are more resistant than HL-60 cells to cell damage induced by BPA and nitro-BPA. As a result of MCF-7 cells exposure to BPA, we saw a slight increase in membrane permeability (250 mM), ROS generation after 24h (25 mM) and increase in cell population in sub G1, so we had DNA fragmentation (100 uM and 250 uM), but with no effect on cell cycle. However, we observed increased levels of CEdG and 8-oxodG on DNA of cells incubated with BPA (100 uM, 250 mM) without prior activation of Ah receptors. The activation of Ah receptors with PCB took a small increase in the level of DNA lesions after incubations with BPA. MCF-7 cells resistance to the cytotoxic effects of BPA is probably related to estrogen action of this compound. Estrogen signaling in addition with the increased levels of DNA damage increases the chance of mutations and malignant transformation. In cells with Ah receptor activation, BPA also led to increased DNA global hydroxymethylation, without changing the global DNA methylation. Nondiabetic animals exposed to BPA had decreased amounts of promyelocytes, blasts and rods in the bone marrow, with any change in blood count. There were an increase of CEdG levels in liver, methylation and global hydroxymethylation on hepatic DNA, and was observed any alteration on epigenetic markers and DNA adducts in kidney or urine. On the other hand diabetic animals exposed to BPA showed increased numbers of eosinophils and lymphocytes in bone marrow, suggesting the induction of an allergic inflammatory state. There were increased levels of CEdG, methylation and global hydroxymethylation on hepatic DNA, increased 8-oxodG levels on kidney DNA without changing epigenetic markers, was not observed DNA adducts in urine. The data obtained indicate that the generation of ROS could be the major route of cytotoxic and genotoxic induced by BPA exposure. BPA biotransformation to BPA-3,4-quinone used in the models seem to have poor effects, since we was not detected BPA-Gua lesion in any DNA sample, culture medium of cells or urine of the animals. Metabolic changes induced by BPA and ROS can enable changes in epigenetic markers observed in the DNA of HL-60 cells, MCF-7 and liver tissue. All these changes may contribute to malignant transformation of cells that were exposed to BPA

Preconditioning with mono and polyunsaturated fatty acids and low-intensity electrical stimulation. Effects on skin repair in rats

PURPOSE: To evaluate the effects of preconditioning with oils mixes containing ω3/ω6/ω9 associated with micro-currents on skin repair in rats. METHODS: One-hundred and eight Wistar rats randomized into G-1, G-2 and G-3 groups were treated with saline (0.9%), mix 1 (corn+soybean oils) and mix 2 (olive+canola+flaxseed oils), respectively, in a single dose (0.01ml/g) by gavage. Next, each group was subdivided into sham and stimulated subgroups. Pulsed-wave microcurrents (0.5 µA, 0.5 Hz) were applied to stimulated subgroups for 20 min. One hour later anesthetized rats were subjected to surgery. A dorsal incision (6 cm long) was carried out and closed with interrupted nylon sutures. Samples (1cm2 ) were harvested from the mid-portion of the incision on the 7, 14, 21 post-operative (P.O.) days. Variables were analyzed using Mann-Whitney/Dunn tests Significance level was set to 5 % (p<0.05). RESULTS: Micro-currents promoted increase of exudate and reduction of epithelialization on day 7 in G1 rats. Mixes 1/2 reduced vascularization on 7/14th days P.O. Both 1/2 mixes reduced fibrosis on day 14. Preconditioning with mix 1 led to increased expression of NF-kB on the 7th day. CONCLUSION: Preconditioning with microcurrents has pro-inflammatory effects while oil mixes 1 and 2 decrease fibrosis and vascularization in the proliferative phase of cicatrization.

A preliminary study of the anti-inflammatory and anti-apoptotic effects of crocin against gastric ischemia-reperfusion injury in rats

The aim of the present study was to investigate the protective effect of crocin on gastric mucosal lesions caused by ischemia-reperfusion (I/R) injury in rats. Thirty-two male rats were randomly divided into sham, I/R, I/R + crocin pretreatment and crocin alone groups. To induce I/R lesions, the celiac artery was clamped for 30 min, and the clamp was then removed to allow reperfusion for 3 h. Crocin-pretreated rats received crocin (15 mg/kg, i.p.) 30 min prior to the induction of I/R injury. Samples of gastric mucosa were collected to quantify the protein expression of caspase-3, an apoptotic factor, and inducible nitric oxide synthase (iNOS), a pro-inflammatory protein, by Western blot. Pretreatment with crocin decreased the total area of gastric lesions and decreased the protein expression levels of caspase-3 and iNOS induced by I/R injury. Our findings showed a protective effect of crocin in gastric mucosa against I/R injury. This effect of crocin was mainly mediated by reducing the protein expression of iNOS and caspase-3.

O objetivo do presente estudo foi investigar o efeito protetor da crocina em lesões da mucosa gástrica causadas por isquemia-reperfusão (I/R) em ratos. Trinta e dois ratos machos aleatoriamente divididos em grupos de ratos normais, operados como controle, I/R. I/R + pré-tratamento com crocina e crocina sozinha. Para induzir lesões I/R, a artéria celíaca foi grampeada durante 30 minutos e, em seguida, o grampo foi removido para permitir a reperfusão por 3 h. Ratos com pré-tratamento com crocina receberam crocina (15 mg/kg, ip) 30 minutos antes da indução do dano I/R. Amostras de mucosa gástrica foram coletadas para qiuantificar a expressão da proteína da caspase-3, o fator apoptótico, e óxido nítrico sintase induzível (iNOS), uma proteína anti-inflamatória, pela técnica de Western Blot. O pré-tratamento com crocina diminuiu a área total de lesões gástricas e a expressão de níveis de caspase-3 e iNOS induzidas pelo dano I/R. Nossos resultados mostraram o efeito protetor da crocina na mucosa gástrica contra o dano I/R. Este efeito foi mediado, principalmente, por diminuição da expressão das proteínas iNOS e caspase-3.

A ação do óleo de pequi (Caryocar brasiliense) no processo cicatricial de lesões cutâneas em ratos / The effect of the Pequi oil (Caryocar brasiliense) in the healing of skin lesions in mice

RESUMO O objetivo deste estudo foi analisar o efeito do óleo de pequi no processo cicatricial de lesões cutâneas em ratos. A pesquisa foi iniciada após a provação da CEUA- FACID sob o nº de protocolo 005/12 e obedeceu aos princípios éticos da experimentação animal de acordo com a Lei Federal nº 11.794/2008. Foram utilizados 20 ratos machos, Wistar (Rattusnorvegicus), peso corpóreo de 300-350g, divididos aleatoriamente em dois grupos iguais: GI- controle (C); GII- tratado com óleo de pequi (T). Cada grupo foi dividido em dois subgrupos, de cinco animais cada, conforme os tempos experimentais estudados de 7(A) e 14(B) dias. Após anestesia e antissepsia foi produzida cirurgicamente ferida circular de 2,5 cm de diâmetro na região dorso lombar do animal. Os animais do Grupo II foram tratados com aplicação tópica diária de 1 ml do óleo de pequi, respeitando os tempos experimentais descritos. Concomitante à mensuração da área da lesão, os ratos foram eutanasiados para realização do processamento histológico e análise do percentual de regressão das lesões. No grupo GII nos diferentes tempos experimentais de 7 e 14 dias foi observado maior percentual de regressão das lesões em relação ao GI (p < 0,05). A partir da análise histológica foi possível detectar que no GII houve menor número de células inflamatórias e maior número de fibroblastos em relação ao GI nos diferentes tempos experimentais (p < 0,001). Conclui-se que o uso do óleo de pequi apresentou influência positiva no processo de reparo de lesões cutâneas em ratos, por promover maior velocidade do reparo tecidual, fato evidenciado pelo fechamento mais rápido das feridas e observação de características inflamatórias reduzidas no grupo tratado em relação ao grupo controle, sugerindo que a inflamação pode já ter regredido no grupo tratado.

ABSTRACT The aim of this study was to analyze the effect of pequi oil in the healing process of skin lesions in rats. The research was initiated after the ordeal of CEUA- FACID with protocol No. 005/12 and following the ethical principles of animal experimentation according to the Federal Law No. 11,794 / 2008. 20 Wistar rats were used (Rattus norvegicus), with body weight ranging from 300-350g. They were randomly divided into two groups: GI control (C) and GII treated with pequi oil (T). Each group was divided into two subgroups of five animals each, according to the experimental study of 7 days (A) and 14 (B) days. After anesthesia and antisepsis, a circular wound of 2.5 cm of diameter was surgically inflicted in the lumbar dorsal region of the animal. The animals in Group II were treated with daily topical application of 1 ml of pequi oil, respecting the described experimental times. Concomitantly with the measurement of the injury area, the rats were euthanized so that a histological processing and a regression analysis of the percentage of injuries could be performed. In GII, at different experimental times of 7 and 14 days, there were a higher percentage of lesions regression compared to GI (p <0.05). From the histological examination it was possible to detect that, at GII, there were a lower number of inflammatory cells and increased number of fibroblasts compared to IM at different time of the trial (P <0.001). It is concluded that the use of pequi oil presented positive influence on the healing process of skin lesions in rats by promoting quicker tissue repair, as indicated by the faster closure of the wounds and the observation of reduced inflammatory characteristics in the group treated compared to the control group, suggesting that the inflammation could have already receded in the treated group.

Reconstruction of abdominal wall defects using small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor

To construct a new biomaterial-small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor, and to evaluate the new biomaterials for the reconstruction of abdominal wall defects. Thirty six Sprague-Dawley rats were used in the animal experiments and randomly divided into three groups. The new biomaterial was constructed by combining small intestinal submucosa with gelatin hydrogel for basic fibroblast growth factor release. Abdominal wall defects were created in rats, and repaired using the new biomaterials (group B), compared with small intestinal submucosa (group S) and ULTRAPROTM mesh (group P). Six rats in each group were sacrificed at three and eight weeks postoperatively to examine the gross effects, inflammatory responses, collagen deposition and neovascularization. After implantation, mild adhesion was caused in groups B and S. Group B promoted more neovascularization than group S at three weeks after implantation, and induced significantly more amount of collagen deposition and better collagen organization than groups S and P at eight weeks after implantation. Small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor could promote better regeneration and remodeling of host tissues for the reconstruction of abdominal wall defects.

Urethral dysfunction due to alloxan-induced diabetes. Urodynamic evaluation and action of sildenafil citrate

To evaluate the effect of diabetes mellitus and of sildenafil citrate on female urethral function. Twenty nine female rats were divided into four groups: G1 - (n=9), normal rats; G2 - (n=6), normal rats treated with sildenafil citrate; G3 - (n=9) rats with alloxan-induced diabetes; G4 - (n=5) rats with alloxan-induced diabetes treated with sildenafil citrate. Under anesthesia, urodynamic evaluation was performed by cystometry and urethral pressure simultaneously. A significant increase in urethral pressure was observed during micturition. Sildenafil citrate can partially reduced urethral pressure in diabetic female rats.

The neuroprotective effect of Sulindac after ischemia-reperfusion injury in rats

To investigate the neuroprotective effects of Sulindac on the hippocampal complex after global cerebral ischemia/reperfusion (I/R) injury in rats. Thirty one Sprague-Dawley rats were used, distributed into group I (sham) n:7 were used as control. For group II (n:8), III (n:8) and IV (n:8) rats, cerebral ischemia was performed via the occlusion of bilateral internal carotid artery for 45 minutes and continued with reperfusion process. 0.3 mL/kg/h 0.9 % sodium chloride was infused intraperitoneally to the Group II rats before ischemia, 5μg/kg/h/0.3 ml sulindac was infused intraperitoneally to the Group III rats before ischemia and 5μg/kg/h/0.3 ml sulindac was infused intraperitoneally to the Group IV rats after ischemia and before reperfusion process. The levels of MDA, GSH and MPO activity were measured in the left hippocampus tissue. The hippocampal tissue of all group members were taken for histopathological study. The MDA and MPO levels increased from group I (control) to group II (I/R) (P<0.05) and decreased from group II (I/R) to group III (presulindac + I/R) and IV (postsulindac + I/R) (P<0.05). Beside these, the GSH levels decreased from group I (control) to group II (I/R) (P<0.05) and increased from group II (I/R) to group III (presulindac + I/R) and IV (postsulindac + I/R) (P<0.05).The number of apoptotic neurons increased from group I (control) to group II (I/R) (P<0.05) and decreased from group II (I/R) to group III (presulindac + I/R) and IV (postsulindac + I/R) (P<0.05). The Sulindac may have neuroprotective effects on ischemic neural tissue to prevent the reperfusion injury after ischemia.

Effects of subcutaneous carbon dioxide on calcitonin gene related peptide and substance P secretion in rat skin

To investigate the subcutaneous injection of carbon dioxide (CO2) on neuropeptides Calcitonin Gene-Related Peptide (CGRP) and Substance P (SP) secretion in rat skin. Fifty-six Wistar-EPM rats were distributed in two groups: one for CGRP analysis, the other for SP analysis. Each group was subdivided into four subgroups: control (Cont), control with needle (ContNd), CO2 injection (CO2Inj) and atmospheric air injection (AirInj) - with seven animals each. Sample analyses of partial skin were conducted by Western Blotting (WB). RESULTS: In SP group, there was a decrease in the amount of neuropeptides in subgroups CO2Inj and AirInj. Similarly, in CGRP group, there was a decrease in the amount of pro-CGRP neuropeptides (15 kDa) in subgroups CO2Inj and AirInj; Nevertheless, there was no decrease in the amount of CGRP (5 kDa) in any subgroups. Subcutaneous injection of CO2 and atmospheric air decreased the amount of Substance P and pro-Calcitonin Gene-Related Peptide (15 kDa) neuropeptides in rat skin.

Ovarian blood vessel occlusion as a surgical sterilization method in rats

To evaluate the female sterilization by occlusion of the ovarian blood flow, using the rat as experimental model. Fifty-five females rats were divided into four groups: I (n=10), bilateral ovariectomy, euthanized at 60 or 90 days; II (n=5), opening the abdominal cavity, euthanized at 90 days; III (n=20), bilateral occlusion of the ovarian blood supply using titanium clips, euthanized at 60 or 90 days; and IV (n=20), bilateral occlusion of the ovarian blood supply using nylon thread, euthanized at 60 or 90 days. The estrous cycle was monitored by vaginal cytology. After euthanasia, the reproductive tissues were evaluated histologically. RESULTS: Ovarian atresia was identified macroscopically at 60 days after surgery in the rats in groups III and IV; however, most of the rats in group III maintained cyclicity. Histology of the tissues from group IV revealed that the ovarian tissue was replaced by dense fibrous connective tissue that was slightly vascularized and that intact follicles were absent by 90 days. OOvarian blood vessels occluded caused ischemia, leading to progressive tissue necrosis, and bilateral occlusion using a nylon ligature is a viable method for surgical sterilization.

Comparative study of tissue reactivity to n-butyl-2-cyanoacrylate and nylon monofilament thread on pericranium-cutaneous flaps in rats

To study the repair of pericranium-cutaneous flaps fixed with suture anchored in a skull bone tunnel or N-butyl-2-cyanoacrylate adhesive in Wistar rats with emphasis on the cellular inflammatory response and the production of types I and III collagen. The operated region in the cephalic region of Wistar rats was removed minutes before euthanasia, fixed in formalin, and subjected to histological preparation. Slides were stained with hematoxylin-eosin and Picrosirius. Standardized counts of polymorphonuclear and mononuclear cells, fibroblasts, and macrophages were performed, and the percentages of types I and III collagen were determined. Data collection occurred on days 3, 7, 14, 21, and 45 postoperatively. A value of p<0.05 was considered statistically significant. Quantitative analysis of the data showed more fibroblasts in the surgical adhesive group than in the nylon monofilament thread groups (p=0.0211). Qualitative analysis showed higher reactivity in the adhesive group, with a predominance of polymorphonuclear cells from days 3-45 and macrophages from days 3-7. The amount of type I collagen exceeded 80% in the treated and control groups at the end of the experiment. CONCLUSIONS: Subperiosteal detachment triggers a cellular inflammatory response that is amplified using soft tissue fixation methods. The adhesive n-butyl-2-cyanoacrylate was more reactive than the nylon monofilament thread anchored in the skull bone tunnel.

Comparative efficacy of Belzer or Euro-Collins solutions for pancreatic preservation during cold ischemic storage in rats

To compare the efficacy of different types of solutions (Belzer or Euro-Collins) for the preservation of rat pancreas during cold ischemia. METHODS: Thirty Wistar rats were divided into three groups according to the perfusion or storage solution: Group E (perfusion and storage in Euro-Collins solution); Group B (perfusion and storage in Belzer solution) and Group BE (Perfusion in Belzer solution and storage in Euro-Collins solution). After perfusion, the pancreas was excised and stored at 4˚C for 18 hours. Amylase was measured at 6, 12 and 18h, and histological analysis of the pancreas was performed after 18h of cold storage. RESULTS: Amylase was elevated and comparable in Groups E and BE after 12 and 18 hours of ischemia (p<0.05). In the exocrine pancreas, histological differences in the amount of necrosis (p=0.049), lymphocytic infiltrate (p<0.001) and neutrophilic infiltrate (p=0.004) were observed, with more favorable features present in Group B. In the endocrine pancreas, Group B showed less edema (p<0.001), but other parameters were similar among all groups. CONCLUSION: The Euro-Collins solution is inferior to the Belzer solution for the preservation of rat pancreas during cold ischemia.

Influence of Annona muricata (soursop) on biodistribution of radiopharmaceuticals in rats

To evaluate the effect of hydroalcoholic extract of A. muricata on biodistribution of two radiopharmaceuticals: sodium phytate and dimercaptosuccinic acid (DMSA), both labeled with 99mtechnetium. METHODS: Twenty four Wistar rats were divided into two treated groups and two controls groups. The controls received water and the treated received 25mg/kg/day of A. muricata by gavage for ten days. One hour after the last dose, the first treated group received 99mTc-DMSA and the second sodium 99mTc-phytate (0.66MBq each group), both via orbital plexus. Controls followed the same protocol. Forty min later, all groups were sacrificed and the blood, kidney and bladder were isolated from the first treated group and the blood, spleen and liver isolated from the second treated group. The percentage of radioactivity per gram of tissue (%ATI/g) was calculated using a gamma counter. RESULTS: The statistical analysis showed that there was a statistically significant decrease (p<0.05) in the uptake of %ATI/g in bladder (0.11±0.01and1.60±0.08), kidney (3.52±0.51and11.84±1.57) and blood (0.15±0.01and 0.54±0.05) between the treated group and control group, respectively. CONCLUSION: The A. muricata hydroalcoholic extract negatively influenced the uptake of 99mTc-DMSA in bladder, kidney and blood of rats.

Validation of the gastroschisis experimental model and the influence of the mother's diet enriched with glutamine in the fetal morphology

To validate the gastroschisis experimental model in female rats and the effects on the glutamine fetal morphology during pregnancy. METHODS: Twelve pregnant rats Wistar were separated in two groups: Group I (n = 6 rats, 71 fetuses) took glutamine and Group II (n = 6 rats, 75 fetuses) took isocaloric supplementation. At the 18th day of pregnancy, female rats were taken to hysterotomy and the fetuses which were selected for the act of gastroschisis were partially removed from the womb and by the laparotomy technique, the exclusion of the intestine was done. After that, fetuses were put in the womb cavity again and the rats' abdomen sutured. At the 21st day of pregnancy, date before delivery, by C-section ordinary animals and the ones with gastroschisis were removed and studied separately. The morphometrical parameters studied were the body weight (PC); the intestine weight (PI); the intestine length (CI) and its relations (PI/PC, PI/CI e PC-PI). RESULTS: The intestine weight (PI) and the intestine length (CI) were different in fetuses with gastroschisis (p<0.05), however no difference between the groups regarding supplementation with glutamine. CONCLUSIONS: The gastroschisis experimental model is valid and reproducible. The nutritional therapy with glutamine did not change the morphometrical parameters.

Oxidative stress and fatty acid profile in Wistar rats subjected to acute food restriction and refeeding with high-fat diets

To assess oxidative stress and the profile of fatty acids incorporated into the hepatic tissue of animals refed with high-fat (HF) diets after acute food restriction. METHODS: Fifty male Wistar rats were divided into five groups and fasting for 48 hours. One group was sacrificed without refeeding (NR), a control group (C) was refed with the standard AIN-93 diet and the remaining groups with HF diets respectively consisting of hydrogenated vegetable oil (PHVO), trans-free (TF) margarine and trans-free margarine enriched with ω-3 and ω-6 (O). After this period the animals were sacrificed for malondialdehyde (MDA), catalase and hepatic fatty acid determination. RESULTS: The groups refed with HF diets showed elevation of MDA levels compared to the C group (p<0.001 for GVH and p<0.01 for TF and O). Hepatic catalase activity was higher in the TF and O groups compared to group C (p<0.05 for both). The amount of saturated fatty acids was lower in the PHVO and O groups compared to the remaining ones (p<0.001). CONCLUSION: The consumption of high-fat diets after prolonged fasting favors oxidative imbalance in hepatic tissue.

Description of a new experimental model skin flap for studying skin viability in rats

To describe a new experimental flap for studying skin viability in rats. METHODS: Twenty male Wistar rats weighing between 250-300g were divided into two groups: group A - McFarlane technique, a 4 x 10cm flap was used (McFarlane); and in group B modified McFarlane technique, a 3 x 10cm flap was used. Seven days later, the animals were sacrificed and the area of necrosis was evaluated in both groups. RESULTS: Group A presented necrosis in 3% of the total area of the flap (CI: 0.01-0.05), Group B presented necrosis in 37% of the total area of the flap (CI: 0.29-0.46), (p<0.001). CONCLUSION: The modified McFarlane flap presented a larger area of necrosis and could be an adequate experimental model of skin flap viability.

Hepatotoxicity and oxidative stress induced by Naja haje crude venom

Snake venoms are synthesized and stored in venom glands. Most venoms are complex mixtures of several proteins, peptides, enzymes, toxins and non-protein components. In the present study, we investigated the oxidative stress and apoptosis in rat liver cells provoked by Naja haje crude injection (LD50) after four hours. Wistar rats were randomly divided into two groups, the control group was intraperitoneally injected with saline solution while LD50-dose envenomed group was intraperitoneally injected with venom at a dose of 0.025 μg/kg of body weight. Animals were killed four hours after the injection. Lipid peroxidation, nitric oxide and glutathione levels were measured as oxidative markers in serum and liver homogenate. In addition, liver function parameters and activities of antioxidant enzymes were determined.