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Propolis has various pharmacological properties of clinical interest, and is also considered a functional food. In particular, hydroalcoholic extracts of red propolis (HERP), together with its isoflavonoid formononetin, have recognized antioxidant and anti-inflammatory properties, with known added value against dyslipidemia. In this study, we report the gastroprotective effects of HERP (50&ndash, 500 mg/kg, p.o.) and formononetin (10 mg/kg, p.o.) in ethanol and non-steroidal anti-inflammatory drug-induced models of rat ulcer. The volume, pH, and total acidity were the evaluated gastric secretion parameters using the pylorus ligature model, together with the assessment of gastric mucus contents. The anti-Helicobacter pylori activities of HERP were evaluated using the agar-well diffusion method. In our experiments, HERP (250 and 500 mg/kg) and formononetin (10 mg/kg) reduced (p &lt, 0.001) total lesion areas in the ethanol-induced rat ulcer model, and reduced (p &lt, 0.05) ulcer indices in the indomethacin-induced rat ulcer model. Administration of HERP and formononetin to pylorus ligature models significantly decreased (p &lt, 0.01) gastric secretion volumes and increased (p &lt, 0.05) mucus production. We have also shown the antioxidant and anti-Helicobacter pylori activities of HERP. The obtained results indicate that HERP and formononetin are gastroprotective in acute ulcer models, suggesting a prominent role of formononetin in the effects of HERP.

Peripheral axon injury and degeneration are often mediated by oxidative stress and inflammation. The hydroalcoholic extract of the red propolis (HERP) has attracted great attention because of its antioxidant and anti-inflammatory activities.The objective of this work is to study the effect of HERP on nerve repair and functional recovery after sciatic nerve injury (SNI) in rats.The chemical markers in HERP were identified using high-resolution mass spectroscopy. After axonotmesis of sciatic nerve, ibuprofen (IBP) and HERP treatments were orally administered for 28 d. Behavioural tests were performed weekly after SNI. The myelinated axon number was counted using morphometric analysis.The compounds found in HERP were pinocembrin, formononetin, vestitol, and biochanin A. The animals that underwent SNI showed a significant decrease in motor function based on the Basso, Beattie and Bresnahan scale and sciatic functional index compared with sham animals until 7 d after the surgery (p 0.05). After 14 and 21 d, the SNI groups treated with either HERP or IBP showed significant improvement (p 0.01), and the SNI group treated with HERP 10 mg/kg showed accelerated motor recovery compared with the other groups (p 0.01). SNI caused also a reduction in the myelinated axon counts, and treatment with HERP 10 mg/kg induced a significant increase in the number of myelinated fibres compared with all other groups.HERP promoted regenerative responses and accelerated functional recovery after sciatic nerve crush. Thus, it can be considered to be a new strategy or complementary therapy for treating nerve injuries.

Propolis has been used as a folk medicine for centuries around the world due to its wide spectrum of biological activities. The red propolis, a new Brazilian variety of this apimaterial, has presented an unusual chemical composition, including isoflavones such as formononetin and biochanin A. Since both the green and red varieties of propolis are traditionally used as medicine and commercialized with no label differentiation, the study of the activities of red propolis extract has become important in order to clarify whether this product has the same activities as commercial ones. In this work, we demonstrated the potential action of the hydroalcoholic extract of red propolis (HERP) and its biomarker, formononetin, as antinociceptive and anti-inflammatory drugs on experimental models.The HERP was chemically characterised by HPLC/DAD analyses. The biological activities of the HERP (3, 10, and 30mg/kg) and formononetin (10mg/kg) were evaluated using the antinociceptive (acetic acid, formalin, and glutamate injections) and anti-inflammatory (carrageenan-induced hindpaw oedema and peritonitis) models in mice after oral administration. The open field test was also performed.Formononetin, one of the main biomarker of red propolis, was identified in the HERP (21.62mg/g). Pretreatment with the HERP (10 and 30mg/kg) and formononetin (10mg/kg) produced reduction (P0.001) in the number of abdominal writhes, but the HERP was more effective (P0.001) than formononetin. In the formalin test, all HERP doses (3, 10, and 30mg/kg, P0.001) inhibited the late phase (inflammatory pain) of formalin-induced licking, but the inhibition of neurogenic pain was observed only when the higher doses (10 and 30mg/kg; P0.05) were used. Formononetin caused inhibition (P0.001) only in the second phase of formalin-induced nociception similarly at all HERP doses in the same phase of the test. The responses in glutamate-induced model presented crescent inhibition (P0.05) with 10 and 30mg/kg of HERP. Also, formononetin inhibited (P0.001) the nociception induced by glutamate similarly to 30mg/kg of HERP. There were no significant differences in the open field test after HERP administration, but formononetin decrease the spontaneous motor behaviour. Regarding the anti-inflammatory assessment, the HERP (10 and 30mg/kg, P0.05) and formononetin (P0.001) treatments caused a significant inhibition of the oedema response. All doses of HERP (3, 10, and 30mg/kg, P0.05) and formononetin (P0.001) also inhibited the carrageenan-induced leukocyte migration. In both cases, the results for the HERP at 30mg/kg and formononetin were similar.The HERP and formononetin presented significant anti-inflammatory activity. Moreover, the HERP presented antinociceptive action on inflammatory and neurogenic pain without motor side effects, possibly due to the action of other constituents present in the extract. These results, together, support the popular usage of this natural product.

The essential oil of Alpinia zerumbet (EOAz) presents myorelaxant and antispasmodic actions on cardiac and smooth muscles. The aim of this study was to investigate the effect of EOAz on the skeletal muscle contraction in post-stroke spasticity. Fifteen adults with unilateral hemiparesis and spasticity resulting from stroke were submitted to surface electromyography readings of the gastrocnemius muscle, before and after 10 daily applications (dermal 0.05 mL per muscle belly) of EOAz. The healthy contralateral muscles without applying the oil were used as controls. The analysis showed that, in both lateral and medial gastrocnemius, the values of all studied variables (root mean square, maximum amplitude and median power frequency) were significantly decreased in pathological legs during muscle contraction (Wilcoxon test, p < 0.05). Moreover, spastic muscles presented different results before and after dermal application of EOAz: The mean values of root mean square and median power frequency were significantly increased in lateral and medial gastrocnemius, and also, the maximum amplitude increased in medial gastrocnemius (Mann-Whitney test, p < 0.05). The results suggest that EOAz acts in the skeletal spastic muscle contraction by promoting relaxation and improvement of the muscular performance. Thus, the EOAz can be useful for the clinical management of secondary effects in patients with cerebral vascular disease.

Purpose: To investigate the cellular response to injury, analyzing histopathologic changes associated with increased cellularity, degeneration and disorganization of collagen fibers. Methods: Thirty wistar rats were divided in two groups after partial Achilles tenotomy: the right hind paw were treated with the essential oil of Alpinia zerumbet (EOAz), diluted to 33% (0.3 mL kg-1), and the left hind paw received sunflower oil for 3, 14, 30 and 90 days. Statistical significance was determined using a Chi-square and Pearson Correlation qualitative variables test. Moreover, Mann-Whitney U-test test for comparison between different groups of the same cell, one-way ANOVA, and Tukey’s test of quantitative measurement. Results: A decrease hyperemia (p < 0.001) was observed in the acute phase of inflammatory cell number (p < 0.001), whereas sub-acute phase was marked by significant correlation with macrophages in fibroblasts (r = 0.17, p = 0.03), with probable induction a dense and modeled tissue. At chronic phase, it was found an increase in the number of fibroblasts and a higher percentage of type I collagen fibers (78%) compared with control collagen fibers (55%). Conclusion: Oil of Alpinia zerumbet stimulated the process of maturation, organization and tissue repair which gave it greater resistance.

Ulcerative colitis (UC) is a common intestinal inflammatory disease with an etiology that is not well understood. Although the anti-inflammatory and anti-oxidant effects of the hydroalcoholic extract of Brazilian red propolis (HERP) have been reported in various experimental models, its protective effect in models of UC have not been evaluated. The purpose of this study was to investigate the chemopreventive effect of hydroalcoholic extract of Brazilian red propolis (HERP) in acetic acid-induced colitis (AAIC) using a rodent model. The HERP was chemically characterised by HPLC/DAD analyses. Male rats were randomly assigned into four groups: sham, vehicle (with AAIC, treated with vehicle), P10 (with AAIC, treated with 10mg/kg HERP), and P100 (with AAIC, treated with 100mg/kg HERP). Treatments were performed for 7days, and colitis was induced on day seven. Animals were euthanized 24h after colitis induction and body weight, colon length, gross and histological scores, malondialdehyde (MDA) and myeloperoxidase (MPO) concentrations in colon tissue, and the immunohistochemical expression of inducible nitric oxide synthase (iNOS) were assessed. The major compounds found in HERP were liquiritigenin (68.8mg/g), formononetin (54.29mg/g), biochanin A (30.97mg/g), and daidzein (19.90mg/g). Rats treated with 10mg/kg HERP demonstrated significant decreases in MPO concentrations, gross and histological scores of tissue damage, and iNOS expression (p

The Portulaca oleracea L. (Portulacaceae) is a cosmopolitan species with a wide range of biological activities, including antioxidant and neuroprotective actions. We investigated the effects of P. oleracea extracts in a 6-hydroxydopamine rat model of Parkinson's disease, a debilitating disorder without effective treatments. Chemical profiles of aqueous and ethanolic extracts of whole plant were analyzed by thin layer chromatography and the antioxidant activity was assessed by 2,2-diphenyl-1-picrilhidrazila method. Male Wistar rats received intrastriatal 6-hydroxydopamine and were treated with vehicle or extracts (oral, 200 and 400 mg/kg) daily for two weeks. The behavioral open field test was conducted at days 1 and 15. Immunohistochemical analysis was performed 4 weeks after surgery to quantify tyrosine-hydroxylase cell counts in the substantia nigra pars compacta. Extracts presented antioxidant activity in concentrations above 300 µg/kg. The chromatographic analysis revealed the presence of Levodopa, alkaloids, flavonoids, saponins, tannins, terpenoids and polysaccharides. Both extracts improved motor recovery 15 days after lesion and protected from tyrosine-hydroxylase cell loss after 4 weeks, but these effects were more evident for the aqueous extract. Because the dopamine precursor is present, in addition to antioxidant compounds and neuroprotective effects, P. oleracea can be considered as potential strategy for treating Parkinson's disease. RESUMO A Portulaca oleracea L. (Portulacaceae) é uma espécie cosmopolita com uma vasta gama de atividades biológicas, incluindo as ações antioxidante e neuroprotetora. No presente trabalho foram investigados os efeitos do tratamento com extratos de P. oleracea em modelo animal (aplicação de 6-hidroxidopamina) para estudo da doença de Parkinson, uma desordem debilitante para a qual não há tratamentos efetivos. Os perfis químicos do extrato aquoso e etanólico obtidos a partir de todas as partes da planta foram analisados por cromatografia em camada delgada e a atividade antioxidante foi avaliada pelo método de 2,2-difenil-1-picrilhidrazil (DPPH). Ratos Wistar machos receberam aplicação intraestrial de 6-hidroxidopamina e foram tratados com veículo ou com os extratos (por via oral, 200 e 400 mg/kg) diariamente durante duas semanas. O teste comportamental de campo aberto foi realizado nos dias 1 e 15. A análise imunohistoquímica foi realizada 4 semanas depois da cirurgia a fim de quantificar as células positivamente marcadas para a enzima tirosina-hidroxilase na substância negra compacta. Os extratos apresentaram atividade antioxidante em concentrações superiores a 300 µg/kg. A análise cromatográfica revelou a presença de levodopa, alcalóides, flavonóides, saponinas, taninos, terpenóides e polissacáridos. Ambos os extratos promoveram recuperação da função motora 15 dias após a lesão e protegeram contra a perda de neurônios positivos para a tirosina-hidroxilase depois de 4 semanas, mas estes efeitos foram mais evidentes para o extrato aquoso. Considerando a presença no precursor dopaminérgico e de compostos com ação antioxidante e neuroprotetora nos extratos, a P. oleracea pode representar uma potencial alternativa para o tratamento da doença de Parkinson.

PURPOSE:To assess the evolution profile of the immunohistochemical expression of stromal constituents over the time-course of wound healing in a murine model.METHODS: Surgical wounds were performed in the back of 24 Wistar rats. After three, seven, 14 and 21 days, six rats were euthanized and the wounded histologically processed to assess the immunohistochemical expression of CD3, CD20, CD31, α-SMA and type-I collagen. Non-injured skin samples (NSS) were used as control. Data were subjected to statistical analysis using ANOVA and Tukey test.RESULTS: The mean of CD3 and CD20 positive cells in the wounds was significantly higher than in NSS at seven and 14 days (p

We investigated the effect of oral administration of hydroalcoholic extract of Brazilian red propolis (HERP) on DMBA-induced oral squamous cell carcinomas (OSCC) in rodents. The chemical components of the HERP were assessed by high-performance liquid chromatography (HPLC). Carcinogenesis was topically induced in the lower lip of 25 rats using 9,10-dimethyl-1,2-benzanthracene (DMBA); the tumour was treated with saline (TUM1) and Tween 80 (TUM2) as well as HERP at 10, 50 and 100 mg/kg (HERP10, HERP50 and HERP100, respectively) for 20 weeks. Topical application of saline and oral administration of 100 mg/kg HERP was used in five rats as a control group (CTR). After 26 weeks, the histological malignancy grading and immunohistochemical expression of Ki-67 and p16(INK4A) were assessed in the tumours/tissue samples. The compounds identified were propyl gallate, daidzein, catechin, epicatechin, formononetin and biochanin A. Formononetin, daidzein and biochanin A showed concentration of 23.29, 0.38 and 0.67 mg/g of HERP, respectively. HERP at doses of 50 and 100 mg/kg inhibited 40% of OSCC growth and promoted a 3-week delay in development of clinically detectable tumours. Epithelial dysplasia was observed in all samples with no clinical tumour, except in CTR. No significant difference in the immunoexpression of Ki-67 and p16(INK4A) was observed between HERP-treated and saline/Tween 80-treated groups (p > 0.05). Our results suggest that HERP exerts chemopreventive activity on the progression of DMBA-induced epithelial dysplasia to OSCC in an experimental model of labial carcinogenesis; however, this effect is not associated with Ki-67 and p16(INK4A) immunoexpression.

PURPOSE: To evaluate the effect of aqueous extract of Hyptis fructicosa on hepatic regeneration after partial hepatectomy in rats. METHODS: Sixteen rats were divided in two groups: C (Control Group) and HF (Whose rats received aqueous extract of Hyptis fructicosa during 4 days using the dose of 100 mg/kg/day). On the consecutive day of this treatment, the animals of both groups underwent hepatectomy of about 67% of liver. Twenty four hours later, they were sacrificed, and the remaining mass of liver was removed and prepared to be studied through the PCNA immunohistochemical technique. RESULTS: The liver regeneration index of HF group was 53.56 ± 18.91%, while in C group was 21.12 ± 8.29% (p=0.0003). CONCLUSION: These results show that the administration of aqueous extract of Hyptis fructicosa using the dose of 100mg/kg/day increased the hepatocyte proliferation in the group HF. OBJETIVO: Avaliar o efeito do extrato aquoso da Hyptis fructicosa sobre a regeneração hepática após hepatectomia parcial em ratos. MÉTODOS: Dezesseis ratos foram divididos em dois grupos: C (grupo controle) e HF (ratos que receberam o extrato aquoso da Hyptis fructicosa durante quatro dias na dose de 100mg/kg/dia). No dia consecutivo deste tratamento, os animais de ambos os grupos foram submetidos a hepatectomia de aproximadamente 67% do fígado. Vinte e quatro horas depois, eles foram sacrificados, e que a massa restante do fígado foi retirado e preparado para ser estudado através da técnica de imuno-histoquímica PCNA. RESULTADOS: O índice de regeneração hepática do grupo HF foi 53,56 ± 18,91%, enquanto no grupo C foi de 21,12 ± 8,29% (p=0,0003). CONCLUSÃO: Estes resultados mostram que a administração do extrato aquoso da Hyptis fructicosa na dose de 100mg/kg/dia aumentou a proliferação de hepatócitos no grupo HF.

Myoepithelioma is a rare benign tumor of the salivary glands and is usually seen in the parotid gland and the minor salivary glands. It was once considered to be a type of pleomorphic adenoma (PA), but myoepitheliomas are today believed to be relatively aggressive tumors. Myoepitheliomas are most common in young adults between the ages of 30 and 50 and there are very few cases reported in individuals less than 18 years of age. We report a case of myoepithelioma located in the hard palate in a 15-year-old Brazilian male. The tumor was composed of plasmacytoid myoepithelial cells. An analysis of the immunohistochemical profile of the tumor cells showed positivity for vimentin, S-100 protein, and glial fibrillary acidic protein (GFAP), but not for smooth muscle actin (α-SMA) and cytokeratin 14 (CK14). We report this case because of the rarity of this tumor, especially in adolescents. We also discuss the histological parameters of the differential diagnosis of this tumor as well as its immunohistochemical profile.

Oxidative stress is involved in many neurodegenerative diseases. Chronic ozone exposure causes a secondary increase of reactive oxygen species, which cause an oxidative stress state in the organism. Ozone is one of the main components of photochemical pollution. Our purpose was to test that oxidative stress caused by chronic low doses of ozone, by itself, alters adult neurogenesis and causes progressive neurodegeneration in the hippocampus, which actions lead to the loss of brain plasticity in the mature central nervous system of rats. Animals were exposed to an ozone-free air stream and for 15, 30, 60, and 90 days to low doses of ozone to cause oxidative stress. Each group was then tested by (1) a spectrophotometer test to quantify lipid peroxidation (LPO) levels; (2) immunohistochemistry testing against doublecortin, Neu-N, p53, microglia, and glial fibrillary acidic protein; (3) Western blot tests for doublecortin and Neu-N; and (4) a one-trial passive avoidance test. Our results indicated that ozone causes an increase of LPO levels, morphological changes in the nucleus and the cytoplasm, and cell swelling in neurons. The Western blot shows a decrease for Neu-N and doublecortin. Activated and later phagocytic microglia and an increased number of astrocytes were found. There was a memory deficiency positively related to the amount of ozone exposure. These alterations suggest that oxidative stress caused by low doses of ozone causes dysregulation of inflammatory processes, progressive neurodegeneration, chronic loss of brain repair in the hippocampus, and brain plasticity changes in the rat analogous to those seen in Alzheimer's disease.

L-DOPA is still the drug of choice to treat Parkinson's disease although adverse side effects appear after several years of treatment. These are thought to be the consequence of plastic re-arrangements of the nigrostriatal connections, such as sprouting of the dopaminergic terminals or post-synaptic changes. Pleiotrophin, a trophic factor that we have shown to be up-regulated in the striatum of parkinsonian rats after long-term L-DOPA treatment may play a role in these plastic changes. To determine whether one of the three known pleiotrophin receptors [N-syndecan, receptor protein tyrosine phosphatase type zeta beta (RPTP-zeta/beta) and anaplastic lymphoma kinase] might be implicated in these putative plastic effects, we quantified their expression levels by real-time RT-PCR in the striatum and mesencephalon of rats with partial lesions of the nigrostriatal pathway undergoing L-DOPA treatment. Both pleiotrophin and RPTP-zeta/beta expression was up-regulated in the striatum but not in the mesencephalon of lesioned rats and RPTP-zeta/beta expression was even further increased by L-DOPA. The levels of the RPTP-zeta/beta protein were also increased in the striatum of L-DOPA-treated lesioned rats. Immunofluorescence labeling showed the protein to be constitutively expressed in striatal medium spiny neurons, which are innervated by both the corticostriatal glutamatergic and nigrostriatal dopaminergic systems. RPTP-zeta/beta might therefore be implicated in the plastic changes triggered by L-DOPA treatment and might merit further study as a potential candidate for Parkinon's disease therapy.

There is evidence that nitric oxide plays a role in the neurotransmitter balance within the basal ganglia and in the pathology of Parkinson's disease. In the present work we investigated in striatal 6-hydroxydopamine (6-OHDA) lesioned rats the effects of a nitric oxide synthase (NOS) inhibitor, NG-nitro-l-arginine (L-NOARG), given systemically on both the dopaminergic (DA) neuronal loss and the neuronal NOS cell density. We analyzed the DA neuronal loss through tyrosine hydroxylase immunohistochemistry (TH). The nitrergic system was evaluated using an antibody against the neuronal NOS (nNOS) isoform. Treatment with the L-NOARG significantly reduced 6-OHDA-induced dopaminergic damage in the dorsal striatum, ventral substantia nigra and lateral globus pallidus, but had no effects in the dorsal substantia nigra and in the cingulate cortex. Furthermore, L-NOARG reduced 6-OHDA-induced striatal increase, and substantia nigra compacta decrease, in the density of neuronal nitric oxide synthase positive cells. These results suggest that nitric oxide synthase inhibition may decrease the toxic effects of 6-OHDA on dopaminergic terminals and on dopamine cell bodies in sub-regions of the SN and on neuronal nitric oxide synthase cell density in the rat brain.

The aim of the present study was to assess degenerative changes in the nitric oxide (NO) system of basal ganglia in animals with experimentally induced Parkinson's disease. In one procedure, rats were stereotaxically injected with 6-hydroxydopamine (6-OHDA) in the right medial forebrain bundle; in a second procedure, electrodes were implanted in the right substantia nigra pars compacta (SNc). After 15 and 30 days animals were tested for rotational asymmetry induced by apomorphine. Apomorphine induced rotation in lesioned animals, towards the ipsilateral side after electrolytic lesion and towards contralateral side in 6-OHDA animals. Structural deficits in basal ganglia were quantified by nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry and by nitric oxide synthase (NOS) immunoreactivity. 6-OHDA and electrolytic lesions induced a significant decrease in the number of NADPH-d/NOS positive cells in the lesion ipsilateral to SNc, in contrast with cell number increase in the ipsilateral dorsal striatum. By contrast, 6-OHDA-treated animals showed a decrease in the number of NOS immunoreactive cells in the contralateral nucleus accumbens. We conclude that populations of NO-synthesizing neurons are differentially regulated in Parkinson's disease induced by different experimental procedures.