Development and characterization of trans-anethole-containing solid lipid microparticles: antiinflammatory and gastroprotective effects in experimental inflammation.

The present study prepared, optimized, and characterized solid lipid microparticles that contained trans-anethole (SLMAN), evaluated their antiinflammatory activity in acute and chronic inflammation models, and investigated their effects on the gastric mucosa in arthritic rats. The microparticles were obtained by a hot homogenization process and characterized by physicochemical analyses. The acute inflammatory response was induced by an intradermal injection of 0.1 ml of carrageenan solution (200 µg) in the hind paw. The rats were treated orally with a single dose of SLMAN 1 h before induction of the inflammatory response. The chronic inflammatory response was induced by the subcutaneous application of 0.1 ml of complete Freund's adjuvant suspension (500 µg) in the hind paw. SLMAN was orally administered, starting on the day of arthritis induction, and continued for 21 days. The results showed that SLMAN was obtained with good encapsulation efficiency. Treatment with SLMAN at doses of 25 and 50 mg/kg was as effective as trans-anethole (AN) at a dose of 250 mg/kg on acute and chronic inflammatory responses. Histological analyses showed that treatment with SLMAN did not aggravate lesions in the gastric mucosa in arthritic rats. These results indicated that treatment with SLMAN at a dose that was 5-10 times lower than non-encapsulated AN exerted an inhibitory effect on acute and chronic inflammatory responses, suggesting the better bioavailability and efficacy of microencapsulated AN without aggravating lesions in the gastric mucosa in arthritic rats.

Prostaglandin E1 prevents histopathological changes improving renal function in experimental nephropathy induced by renal microembolism.

This study investigated the effect of prostaglandin E1 (PGE-1) treatment on the biochemical and histopathological changes in a model of nephropathy that was induced using renal microembolism in rats. Wistar rats were assigned to three groups: a control group (C, normal), a renal microembolism (RM) group, and a renal microembolism treated with PGE-1 (RM + PGE-1) group. The renal microembolism was induced by an arterial injection of polymethylmethacrylate microbeads into the remaining kidney of nephrectomized rats. Intramuscular treatment with PGE-1 was initiated on the day of the induction of the renal microembolism and continued once weekly for up to 60 days. At the end of the treatment period, blood samples were taken to assess the serum creatinine and urea concentrations, and 24-h urine samples were collected to determine the total protein levels. The rats' kidneys were removed and processed for histopathological analysis using the hematoxylin and eosin, periodic acid-Schiff, Mallory-Azan, and Picro-Sirius techniques. An immunohistochemical assay with vascular endothelial growth factor receptor-2 (anti-VEGFR-2) was also performed. The results showed that the PGE-1 treatment prevented vascular, glomerular, tubular, and interstitial alterations and reduced the biochemical changes, thus improving the renal function in rats that were subjected to renal microembolism. These effects could be partially attributable to an increase in the PGE-1-induced angiogenesis, because we observed an increase in the tissue expression of VEGFR-2, a specific marker of angiogenesis.

Anti-inflammatory Effect of Low-Dose Anethole and Ibuprofen Combination Is Accompanied by Partial Prevention of Hepatic Metabolic Changes in Arthritic Rats.

Anethole (AN) is a natural compound that has attracted great scientific interest because of its numerous biological activities, including anti-inflammatory effects. However, these effects were obtained with high doses of AN, which may be one limitation of its therapeutic use. This study evaluated the effects of a low-dose AN and ibuprofen (IB) combination on inflammatory parameters in Freund's complete adjuvant-induced arthritis (AIA) and arthritis-induced hepatic metabolic changes. Holtzman rats were used and divided into groups: normal, AIA (control), arthritics treated with IB, arthritics treated with AN, and arthritics treated with AN + IB. The volume of the paws, the appearance of secondary lesions, and the number of synovial leukocytes were evaluated. Gluconeogenesis and ureagenesis from alanine were determined in the rat liver in isolated perfusion. The AN + IB (62.5 + 8.75 mg/kg) treatment exerted an inhibitory effect on inflammatory parameters and partially prevented hepatic metabolic changes that was similar to the effect of high-dose IB (35 mg/kg) and AN (250 mg/kg) treatment. This effect of the treatments on hepatic metabolism can be, partly at least, explained by the preservation of both the alanine aminotransferase (ALT) activity and the cytosolic NADH/NAD+ redox potential in the liver. Taken together, the data obtained provided evidence that the AN + IB combination at lower doses than AN and IB treatment alone had beneficial inhibitory potential for the treatment of AIA and attenuated metabolic changes in the liver. Graphical Abstract.

Toxoplasma gondii causes lipofuscinosis, collagenopathy and spleen and white pulp atrophy during the acute phase of infection.

In this study, we evaluated homeostatic and functional disorders of the spleen in mice inoculated with Toxoplasma gondii. The kinetics of megakaryocyte and leukocyte production, body and spleen mass and certain histopathological aspects were analyzed. There was increased (P < 0.05) the accumulation of lipofuscin in the red pulp of the spleen, in the periods of 30 and 60 dpi of the infection, that is, in the chronification stage of the disease and decrease of the white pulp area. In addition, we observed (from 7dpi) a quantitative and qualitative increase (P < 0.05) in the deposition of collagen fibers in the spleen of all infected mice. Since resolution of the inflammatory process resulted in pathophysiological changes, we can suggest that the T. gondii invaded and multiplied in the cells of the white and red pulps of the spleen. Although we did not find the parasite in the spleen, this hypothesis is supported by the presence of diffuse inflammatory infiltrate, which extended through the spleen parenchyma of all inoculated mice. Taken together, our results suggest that T. gondii causes severe homeostatic disorders that have altered spleen physiology, including diffuse parenchymal inflammation, lipofuscinosis in histiocytes, early aging, collagenopathy, systemic sclerosis and spleen and white pulp atrophy.

Nephropathy induced by renal microembolism: a characterization of biochemical and histopathological changes in rats.

The aim of this study was to investigate some biochemical parameters of renal function and the vascular, glomerular, tubular, and interstitial manifestations in the progression of nephropathy induced by renal microembolism. Renal microembolism was induced by the arterial injection of polymethacrylate microspheres in the remnant kidney of nephrectomized rats. Animals 110-120 days old were randomly divided into three groups: the control group (C; normal), the nephrectomized group (S; nephrectomized that did not undergo renal microembolism), and the model group (M, nephrectomized animals that underwent renal arterial microembolism). The animals were evaluated 30, 60, and 90 days after the induction of a renal microembolism. Blood and urine samples were collected to determine serum creatinine (Cr) and urea (Ur) concentrations and urine total protein (Pt) concentrations. The kidneys were weighed and processed for histopathological analysis using hematoxylin and eosin (HE), periodic acid-Schiff (PAS), Mallory-Azan, and Picro-Sirius staining. The samples were also subjected to immunohistochemistry with a proliferating cell nuclear antigen (PCNA) and a vascular endothelial growth factor receptor (VEGFR). The data demonstrated evidence of the occurrence of vascular, glomerular, tubular, and interstitial abnormalities in the renal tissue, and changes in the biochemical parameters of renal function (serum Cr and Ur and of 24-h urine Pt) in this experimental model of nephropathy induced by renal microembolism, which may indicate the development of chronic kidney disease (CKD). Additionally, the findings indicate that this is a good reproducibility model that may be useful for studying the pathogenesis of CKD that is caused by atheroembolism and possible treatment alternatives.

Acute Toxoplasma gondii infection alters the number of neurons and the proportion of enteric glial cells in the duodenum in Wistar rats.

BACKGROUND: Toxoplasma gondii infection can occur through the ingestion of raw meat that contains tissue cysts or food that contains oocysts. Through the ingestion of oocysts, the parasite crosses the intestinal barrier, where the enteric nervous system is located. The objective was to investigate the kinetics of neuronal and glial responses during acute T. gondii infection. METHODS: We used 45 Wistar rats that were divided into a control group and infected groups that were evaluated at 6, 12, 24, 48, 72 hours, 7 days, 10 days, and 15 days after infection. The rats received 5000 sporulated oocysts of the parasite orally. To detect neurons and enteric glia cells, the myenteric and submucosal plexuses of the duodenum underwent double-labeling immunohistochemical techniques to evaluate HuC/HuD and S100, HuC/HuD and ChAT, and HuC/HuD and nNOS. KEY RESULTS: We observed a reduction of the total neuron population in the submucosal plexus 72 hours after infection. Cholinergic neurons decreased in the submucosal plexus 15 days after infection, and nitrergic neurons decreased in the myenteric plexus 72 hours after infection. A decrease in the number of glial cells was observed 7 days after infection in the submucosal plexus, and an increase in the enteric glial cell (EGC)/neuron ratio was found in both plexuses 48 hours after infection. CONCLUSIONS AND INFERENCES: We found decrease of neurons and increase in the EGC/neuron ratio in both plexuses caused by acute T. gondii infection, with major alterations 72 hours after oral infection. The number of cholinergic neurons decreased in the submucosal plexus, and the number of nitrergic neurons decreased in the myenteric plexus. A decrease in the number of enteric glial cells was observed in the submucosal plexus, and an increase in the enteric glial cell/neuron ratio was observed in both ganglionate plexuses of the duodenum.

Toxic effect of commercial formulations of neem oil, Azadirachta indica A. Juss., in pupae and adults of the sugarcane borer, Diatraea saccharalis F. (Lepidoptera: Crambidae) / Efeito tóxico de formulações comerciais de óleo de neem, Azadirachta indica A. Juss., em pupas e adultos da broca da cana-de-açúcar, Diatraea saccharalis F. (Lepidoptera: Crambidae)

To evaluate the toxic effect of commercial formulations of neem oil, Azadirachta indica A. Juss, pre-pupae (PP), young pupae (YP) and old pupae (OP) of Diatraea saccharalis F. (Lepidoptera: Crambidae) were sprayed with the diluted extract in distilled water at concentrations of 0.0, 0.3, 0.5, 1.0 and 2.0%. The neem extract caused concentration-dependent effects on mortality of pupae, and the pupae that failed to emerge in adults had multiple abnormalities. The longevity of pupae that emerged in adults (YP and OP group) did not differ from the control group. The abnormalities found in adults were related to mortality in all treatments, except at the concentration of 1.0%. Fertility was assessed according to the oviposition of adult females from the YP group that showed no abnormalities, through the evaluation of the number of deposited eggs and the rate of undeveloped eggs. The results showed a reduction in the number of eggs laid and an increase in the percentage of undeveloped eggs. These results show that neem oil has a high potential to control the toughest stage of the sugarcane borer and also reduces the further development. Therefore, commercial formulations of neem oil have a toxic effect on pupae and adults of D. saccharalis.(AU)

Para avaliar o efeito tóxico de formulações comerciais de óleo de neem, Azadirachta indica A. Juss, pré-pupas (PP), pupas jovens (PJ) e pupas velhas (PV) da Diatraea saccharalis F. (Lepidoptera: Crambidae) foram pulverizadas com o extrato diluído em água destilada, em concentrações de 0,0, 0,3, 0,5, 1,0 e 2,0%. O neem provocou diferentes efeitos sobre a mortalidade de pupas, dependendo da concentração. As pupas que não conseguiram emergir em adultos apresentaram anormalidades múltiplas. Quanto às pupas que emergiram em adultos (grupos PJ e PV), foi calculada a sua longevidade, que não diferiu da do grupo controle. As anormalidades encontradas em adultos estão relacionadas com a mortalidade em todos os tratamentos com exceção da concentração de 1,0%. A fecundidade foi avaliada de acordo com a oviposição de adultos fêmeas do grupo PJ, que não apresentaram anormalidades; dentro dos ovos depositados foi avaliado o número de ovos não desenvolvidos. Os resultados demonstraram redução no número de ovos depositados e aumento na porcentagem de ovos não desenvolvidos. Esses resultados mostraram que o óleo de neem tem elevado potencial para o controle do estágio mais resistente da broca da cana-de-açúcar, além de reduzir o aparecimento das fases subsequentes. Portanto, formulações comerciais de óleo de neem apresentam um efeito tóxico em pupas e adultos de D. Saccharalis.(AU)

Resveratrol promotes myenteric neuroprotection in the ileum of rats after ischemia-reperfusion injury.

AIMS: The present study evaluated the effects of resveratrol in the myenteric plexus after intestinal ischemia-reperfusion (I/R) injury caused by occluding the superior mesenteric artery for 45min, followed by 7days of reperfusion. MAIN METHODS: Forty-two male Wistar rats were divided into seven groups: control (C group), untreated sham surgery control (SC group), sham surgery control treated with resveratrol before surgery (STA group), sham surgery control treated with resveratrol before and after surgery (STAD group), ischemic control (IRC group), ischemic treated before I/R (IRTA group), and ischemic treated before and after I/R (IRTAD group). Resveratrol (10mg/kg) was administered for 4days and 2h prior to surgery and/or 7days later. Morphometric analyses were performed, and the density of the general neuronal population (HuC/D-immunoreactive [IR]), nitrergic subpopulation (neuronal nitric oxide synthase [nNOS]-IR), vasoactive intestinal peptide (VIP)ergic varicosities (VIP-IR), and glial cells (S100-IR) was determined. KEY FINDINGS: Injury that was caused by I/R significantly reduced (p<0.01) the HuC/D-IR general neuronal population. Treatment with resveratrol before and after ischemia had a neuroprotective effect. Morphometric changes caused by I/R in nitrergic neurons and varicosities were also attenuated by resveratrol. Ischemia/reperfusion promoted the proliferation of enteric glial cells, and resveratrol treatment before and after I/R reversed this effect. SIGNIFICANCE: Resveratrol had neuroprotective effects, showing promise for application in intestinal surgery and transplants.

Trypanosoma cruzi (CEPA CL) promove perda neuronal no plexo mioentérico do cólon de ratos / Trypanosoma cruzi (STRAIN CL) causes neuronal loss in rats´ myo-enteric plexus of the colon

Trypanosoma cruzi, um protozoário parasita, é o agente etiológico da doença de Chagas. A infecção chagásica apresenta duas fases: aguda e crônica. Na fase aguda a mortalidade é mais frequente, sendo que o organismo que consegue sobreviver a ela passa para a fase crônica indeterminada. No trato gastrointestinal o T. gondii pode parasitar o sistema nervoso intramural formando os megaesôfago e o megacólon. Não existem trabalhos que relacionam a perda neuronal do plexo mientérico com a infecção com a cepa CL do T. cruzi. Portanto, o objetivo deste trabalho foi verificar alterações morfoquantitativas no plexo mioentérico no colo distal de ratos infectados com a cepa CL durante a fase aguda e crônica da doença. Foram utilizados 20 ratos divididos em quatro grupos: infectado com a cepa CL durante 7 dias (grupo IA) do T. cruzi e seu controle (grupo CA), ambos sacrificados após sete dias do início do experimento, outro grupo infectado (IC) e sacrificado após trinta dias e seu controle (CC). Foi realizada a estimativa da densidade neuronal e a morfometria das áreas do perfil dos corpos celulares dos neurônios através da técnica de Giemsa. A análise quantitativa demonstrou uma redução significativa no número de neurônios nos grupos infectados IA e IC. Em relação ao perfil neuronal foi observada uma atrofia dos neurônios do grupo IC em relação ao controle. Portanto, a cepa CL do T. cruzi provoca uma denervação do plexo mioentérico do colo sem contudo ocasionar hipertrofia neuronal no tempo experimental de trinta dias.

The parasite protozoa Trypanosoma cruzi is the etiological agent of Chagas´s disease. Infection comprises the acute and chronic type. Mortality in the former is more frequent and the organism that survives passes to the undetermined chronic phase. T. gondii within the gastrointestinal tract parasites the intramural nervous system and causes mega-esophagus and mega-colon. No reports in the literature relate neuronal loss of the myo-enteric plexus with infection by strain CL of T. cruzi. Current analysis verifies morphoquantitative alterations in the myo-enteric plexus in rats´ distal colon infected with strain CL during the acute and chronic phase of the disease. Twenty rats were divided into four groups: infected with strain CL of T. cruzi during 7 days (Group IA) and its control (Group CA), killed after seven days from the start of the experiment; another infected group (IC), killed after 30 days and its control (CC). Neuronal density and morphometry of the areas of cell bodies of neurons were estimated by Giemsa technique. Quantitative analysis showed a significant decrease in the number of neurons in infected groups IA and IC. In the case of the neuronal profile, atrophy of neurons of group IC was reported when compared to control. Strain CL of T. cruzi causes a de-nerving of the myo-enteric plexus of the colon without causing a neuronal hypertrophy during the thirty days of experimental period.

Ginkgo biloba (EGb 761) extract: effects on the myenteric plexus of the large intestine in Wistar rats.

The objective of this study was to evaluate the effect of the purified extract of the Ginkgo biloba (EGb 761) plant on the myenteric plexus in the proximal and distal colon of Wistar rats for a period of 120 days. The experimental rats were divided into two age groups: a young group, sacrificed at age 90 days, and an adult group, sacrificed at age 210 days. We observed a significant reduction in the number of neurons in the myenteric plexus of the adult group compared to the young group in both of the segments studied (P < 0.01). The adult group treated with Ginkgo biloba showed a significant increase in neuronal profile area in both the segments studied (P < 0.001). It can be concluded from these results that treatment with the purified Ginkgo biloba (EGb 761) plant extract at a dose of 50 mg/kg body weight has neurotrophic effect on the myenteric plexus in the proximal and distal colon of rats after 120 days of treatment.

Evaluation of the effect of Ginkgo biloba extract (EGb 761) on the myenteric plexus of the small intestine of Wistar rats.

BACKGROUND: The aging process causes a reduction in the myenteric neuronal population, related to oxidative stress, resulting in malfunctioning of the digestive tract. The purpose of this study was to evaluate the action of Ginkgo biloba extract (EGb 761), an important antioxidant drug, on the myenteric plexus of the jejunum and ileum of rats after treatment for 120 days. METHODS: Fragments of the jejunum and ileum were collected from three groups of rats: a 90-day-old group (group Y), a 210-day-old group (group A), and a 210-day-old group treated daily with the extract EGb 761 (50 mg/kg body weight) (group TA). The analysis was carried out by using the myosin-V immunohistochemical technique. Neuronal densities were estimated, and a study of the neuronal profile area of 500 neurons from each group was carried out. RESULTS: In the jejunum, there was a significant neuronal population reduction of 17% only in group A compared with group Y. In the ileum, there was a significant neuronal reduction of 36% in group A compared with group Y, and a significant reduction in group TA of 20%. The difference in the reduction between groups A and TA in the ileum was also significant. In the jejunum, only group A showed a significant increase in neuronal profile area, but in the ileum, there was a significant increase in both groups A and TA. CONCLUSIONS: A daily dose of 50 mg/kg body weight of Ginkgo biloba extract has a significant neuroprotector effect on the myenteric plexus of the ileum during the aging process in rats.