Pouteria ramiflora leaf extract on emulgel in wound healing activity in diabetic rats / Ação do extrato de folhas de Pouteria ramiflora em emulgel na atividade de cicatrização de feridas em ratos diabéticos

The genus Pouteria has been studied because it presents various activities, among which is its anti-inflammatory potential. The effects of Pouteria ramiflora Carbopol gel on the healing of skin wounds in diabetic rats were evaluated by microscopic imaging. Streptozotocin was administered intraperitoneally in animals that had fasted for 12 hours, a situation confirmed by the glycemic index (> 240 mg dL-¹). An excision on the back of the animals was performed and three groups were formed: Control (Gel), Ethanolic extract (Ext) and Gel + extract 2% (Ext+gel); the histopathological evaluation occurred on the 7th, 14th, 21st and 30th days after the post-operative period. The results of the phytochemical prospecting of P. ramiflora extract demonstrated the major presence of phenolic compounds and flavonoids; the assessment of the inflammatory infiltrate on the 7th day was higher on group Ext and Ext+gel when compared to group Control; on the 14th day control and Ext (p<0.05). The quantification of fibroblasts was higher on the 7th day among the three treatments, control and Ext (p<0.05), on the 21st day. Angiogenesis showeda higher number of vessels in Ext+gel group (p<0.05) on the 7th day; in Control, Ext and Ext+gel (p<0.05) on the 14th day; and Control and Ext (p<0.05)on the 21st day. The histopathological results showed that the formulation Ext+gel was efficient in tissue reparation and decrease in inflammatory cells on the diabetic's animals.

O gênero Pouteria apresenta várias aplicações terapêuticas e, dentre elas, grande potencial antiflamatório. Os efeitos do gel de Pouteria ramiflora sobre a cicatrização de feridas na pele de ratos diabéticos foram avaliados pela histomorfometria. A estreptozotocina foi administrada por via intraperitoneal em animais após jejum de 12 horas, a confirmação de indução da diabetes foi confirmada pelo índice glicêmico (> 240 mg dL-1). Foi realizada uma incisão no dorso do animal e foram criados 3 grupos de tratamento: controle (gel carbopol), extrato etanólico (Ext) e Gel + extrato etanólico à 2% (Ext+gel); a avaliação histopatológica foi realizada no 7º, 14º, 21º e 30º dias após o período pós operatório. Os resultados da prospecção fitoquímica dos extratos de P. ramiflora demonstraram majoritariamente a presença de compostos fenólicos e flavonóides; o infiltrado inflamatório avaliado no 7º dia foi maior para animais do grupo controle em relação aos grupos Ext (p<0.05) e Ext+gel 2% (p<0.05); no 14º dia o controle e Exp (p<0.05) apresentaram aumento significativo dos infiltrados inflamatórios. A presença de fibroblastos foi elevada no 7º dia em todos os tratamentos. O processo da angiogênese mostrou um maior número de vasos sanguíneos entre os grupos Ext e Ext+gel (p<0.05) no 7º dia; no 14º dia o grupo controle, Ext (p<0.05), Control e Ext+gel (p<0.05) apresentaram aumento de vascularização, e no 21º dia apenas os grupos controle e Ext (p<0.05). Os resultados histopatológicos mostraram que a formulação gel carbopol + extrato etanólico a 2% foi eficiente na reparação de tecidos e na diminuição de células inflamatórias nos animais diabéticos.

Impact of Diet Containing Grape Pomace on Growth Performance and Blood Lipid Profile of Young Rats.

Grape pomace (GP), the residue of grapes after wine making, is rich in dietary polyphenols and fiber, and it has potential to serve as a functional food ingredient to improve health. However, high polyphenol diets have also been reported to inhibit the growth of young animals and cause liver necrosis. This study investigated the effect of diets containing different amounts of GP on the growth performance and blood lipid profile by using a young rat model. Twenty female Sprague-Dawley rats of age 7 weeks were randomly divided into four groups that were fed AIN-93G diets that were modified by substituting 0%, 10%, 20%, and 30% of carbohydrate with GP for 10 weeks (the diets, thus, obtained contained 0%, 6.9%, 13.8%, and 20.7% of GP). The group fed original AIN-93G (0% GP) was used as control. Feed consumption, body weight, length, and height were recorded weekly. Blood samples were taken biweekly to analyze plasma lipid profile. At the end of the feeding period, the rats were fasted overnight and euthanized by exsanguination under anesthesia. Livers, hearts, and kidneys were collected, and their weights were recorded. Results show that the diet containing a maximum of 20.7% of GP did not influence the body weights, lengths, and heights of rats. As the GP content increased, the blood triglyceride and very low-density lipoproteins (VLDL) decreased, the high-density lipoprotein (HDL) and low-density lipoprotein (LDL) increased slightly but were statistically significant, and total cholesterol remained constant. In conclusion, GP in the AIN-93G diet did not influence the growth performance of young rats, but it exhibited both positive and negative effects on the blood lipid profile.

Reference intervals for biochemical blood variables, packed cell volume, and body temperature in pet rats (Rattus norvegicus) using point-of-care testing.

BACKGROUND: Rats (Rattus norvegicus) are increasingly kept as pets, thus more and more requiring veterinary care. Important differences exist between pet and laboratory rats, but little information is available on pet rats. Diagnosis and treatment of pet rat diseases would benefit from RIs specific for this animal population. OBJECTIVES: The aim was to establish RIs for biochemical blood variables, PCV, and body temperature in pet rats. Additionally, influences of sex and feed rationing method (ad libitum, restricted) on these variables were determined. METHODS: American Society for Veterinary Clinical Pathology (ASVCP) recommendations were followed. Biochemical variables were measured using an automated chemistry analyzer, the VetScan VS2. Nonparametric statistical methods were used to determine RIs and effects of sex and feed rationing method. RESULTS: Reference intervals were established using values of 123 clinically healthy pet rats (except for globulins and albumin/globulin ratio, n = 120) and were: glucose 6.6-13.7 mmol/L, total proteins 66-88 g/L, albumin ≥ 38 g/L, globulins 12-35 g/L, albumin/globulin ratio 1.0-4.7, urea nitrogen 2.5-6.6 mmol/L, creatinine ≤ 53 µmol/L, total bilirubin 4-7 µmol/L, ALP 40-442 IU/L, ALT 22-137 IU/L, amylase 502-1113 IU/L, sodium 133-144 mmol/L, potassium 3.6-5.3 mmol/L, calcium 9.5-10.9 mg/dL, phosphorus 2.3-7.0 mg/dL, PCV 40-50%, and body temperature 35.8-39.3°C. Sex significantly affected 10 variables. No significant influence was found with feed limitation CONCLUSIONS: Reference intervals reported in this study will be useful for interpretation of biochemistry analysis in pet rats and therefore improve pet rat medicine.

Fecal dehydroepiandrosterone (DHEA) immunoreactivity as a noninvasive index of circulating DHEA activity in young male laboratory rats.

Evidence suggests that dehydroepiandrosterone (DHEA) plays a key role in stress and coping responses. Fecal sampling permits assessment of hormone-behavior interactions reliably and effectively, but no previous study has compared circadian- or stress-dependent alterations between serum DHEA and its fecal metabolites. In the current study, young (28 d of age) male rats were assigned to either an experimental (n = 6) or control (n = 6) group. Rats in the experimental group were exposed to a forced swim test to assess their behavioral and physiologic response to an environmental stressor; blood samples were drawn before the test (baseline), immediately after the test, and at 2 later time points. Only fecal samples were collected from control animals. Fecal DHEA and corticosterone metabolites were monitored in all animals for 24 h. DHEA metabolites in control rats exhibited significant diurnal variation, showing a similar temporal pattern as that of corticosterone metabolites. In addition, fecal and serum DHEA levels were highly correlated. Significant peaks in both DHEA and corticosterone metabolite levels were detected. These data suggest that measures of fecal DHEA can provide a complementary, noninvasive method of assessing adrenal gland function in rats.

Circulating leptin levels in newborn rats: a significant post- natal developmental effect, independent of dietary polyunsaturated fat levels.

Leptin expression exhibits developmental and dietary regulation, but it is unknown whether there is an interaction of the regulation by dietary fat and postnatal development. The purpose of this study was to test the effect of different levels of dietary polyunsaturated fat on circulating leptin levels at different post-natal developmental stages. Pregnant (Sprague-Dawley) rats consumed from day 15 of pregnancy through day 9 of lactation a low fat, (11% of energy; LF) polyunsaturated safflower oil diet. From day 9 of lactation, dams and their respective pups were fed low, moderate (40% of energy; MF) or high (67% of energy; HF) polyunsaturated safflower oil diets to full maturation (56 days). Diets were iso-energetic and iso-nitrogenous. Milk fatty acid content reflected the mothers and pups diet, with 15 to 100 fold less C10:0 and 2.6 to 3.3 fold more C18:2 in MF and HF groups compared to LF diet. In newborn rats through post-natal day 56, levels of polyunsaturated fat in mothers' milk and mothers/pups diet had no effect on the levels of circulating leptin. The post-natal development period significantly affected circulating leptin levels (p < 0.001, 15 days = 56 days > 21 days > 28 days). In summary, the developmental postnatal stage regulates leptin levels, independently of the polyunsaturated fat levels in the diet.

An in vivo model for elucidation of the mechanism of tumor necrosis factor-alpha (TNF-alpha)-induced insulin resistance: evidence for differential regulation of insulin signaling by TNF-alpha.

Tumor necrosis factor-alpha (TNF-alpha) has been shown to induce insulin resistance in cultured cells as well as in animal models. The aim of this study was to map the in vivo mechanism whereby TNF-alpha contributes to the pathogenesis of impaired insulin signaling, using obese and lean Zucker rats in which TNF-alpha activity was inhibited through adenovirus-mediated gene transfer. We employed a replication-incompetent adenovirus-5 (Ad5) vector to endogenously express a TNF inhibitor (TNFi) gene, which encodes a chimeric protein consisting of the extracellular domain of the human 55-kDa TNF receptor joined to a mouse IgG heavy chain. Control animals consisted of rats infected with the same titer of adenovirus carrying the lac-z complementary DNA, encoding for beta-galactosidase. There was a significant reduction in plasma insulin and free fatty acid levels in TNFi obese rats 2 days following Ad5 administration. The peripheral insulin sensitivity index was 50% greater, whereas hepatic glucose output was completely suppressed during hyperinsulinemic glucose clamps in TNFi obese animals, with no differences observed between the two lean groups. The improvement in peripheral and hepatic sensitivity to insulin seen in the obese animals was independent of insulin receptor (IR) number and insulin binding affinity for IR. However, TNF-alpha neutralization led to a 2.5-fold increase in tyrosine phosphorylation of IR in skeletal muscle, whereas this was unchanged in liver. There was also a 4-fold increase in particulate protein tyrosine phosphatase activity of skeletal muscle in TNFi obese animals vs. beta-galactosidase controls, whereas protein tyrosine phosphatase activity in liver was unchanged. These results suggest that TNF-alpha is a mediator of insulin resistance in obesity and may modulate IR signaling in skeletal muscle and liver through different pathways. TNF-alpha may affect insulin action in the liver either at sites distal to the IR or indirectly, possibly because of increased provision of gluconeogenic substrates or altered counterregulation. In addition, the Ad5-mediated gene delivery system employed here provides an in vivo model that is efficient and economical for exploring mechanisms involved in TNF-alpha-induced insulin resistance in various genetic models of obesity-linked diabetes.

Effect of dietary calcium on in vitro aortic tissue responsiveness to a hypertensive factor.

It has been proposed that calcium supplementation in the diet is associated with a reduction in blood pressure. In the present study, we investigated vascular tissue sensitivity to a hypertensive factor (HF) in spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY) fed a high calcium diet, a low calcium diet and a food restricted diet. HF, which has been isolated from erythrocytes, increases blood pressure when injected into normotensive rats and stimulates calcium uptake by aortic rings in vitro. Five-week-old rats were divided into the following groups: SHR and WKY fed a regular diet (1% calcium), SHR and WKY fed a high calcium diet (4% calcium), SHR and WKY fed a low calcium diet (0.02% calcium) and SHR and WKY fed a regular diet (1% calcium) in which food intake was restricted to 65% of ad libitum intake. Food intake, body weight, urine phosphate excretion and blood pressure development were followed for 8 weeks. At sacrifice, plasma levels of calcium and phosphate were determined. Tissue responsiveness to HF was calculated by incubating aortic rings from the rats in the different groups with HF and measuring lanthanum-resistant calcium uptake. A 4-fold increase in dietary calcium reduced blood pressure and tissue responsiveness to HF in SHR. Neither parameter was affected by the high calcium diet in WKY. The low calcium diet had no effect on either blood pressure or tissue responsiveness to HF in SHR or WKY. Restriction of food intake induced a reduction in blood pressure and in tissue responsiveness to HF in SHR. It did not affect the same parameters in WKY. The results suggest that the increased tissue responsiveness to HF in the SHR may be associated with high blood pressure.

A comparison of the response of woodchucks and rats to variations in dietary lipotrope and lipid content.

A soy protein-based experimental diet for woodchucks (Marmota monax) is described. The diet supported growth of juvenile woodchucks for 12 wk. With this diet, the effects on both woodchucks and rats of increasing dietary corn oil from 5 to 15% and of deleting supplemental lipotropic factors (choline, methionine, folic acid and vitamin B-12) were studied in a 2 X 2 factorial experiment. Both increased lipid and lipotrope deletion resulted in decreased growth in rats, but only increased lipid caused growth depression in woodchucks. Lipotrope depletion resulted in elevated serum markers of hepatic injury and hepatic lipid accumulation in rats but not in woodchucks. Hematological changes induced by the low lipotrope diets included decreased packed cell volume, total hemoglobin and mean corpuscular volume (MCV) in rats but increased MCV in woodchucks. The woodchuck appears to be more resistant than the rat to induction of hepatic injury by lipotrope deficiency.

[Growth and hematological criteria of rats by different protein and iron supply]. / Wachstum und hämatologische Kriterien von Ratten bei unterschiedlicher Protein- und Eisenversorgung.

In a two-factorial experiment with growing rats, protein content of diet (5, 25, 45%) and Fe supply (5, 25, 625 mg/kg diet) were changed. Both factors as well as their interactions influenced growth (p less than 0.001). The growth was reduced especially by deficient protein supply but although by inadequate iron supply and in a smaller degree by an excessive protein content of the diet. Hematological values as hemoglobin content, counts and volume of erythrocytes, hematocrit, MCH and MCHC - measured after 35 days of the experiment - were influenced by both factors and their interactions, too (p less than 0.001). Again deficient protein supply and insufficient Fe supply have the marked effects. Referring life weight as well as hematological parameters, the deficient protein supply was - independent of Fe supplementation - the limited factor, whereas in the cause of higher protein content (25, 45%) an insufficient Fe supply has negative effects.

Blood selenium and glutathione peroxidase activity in pregnant women: comparative assays in primates and other animals.

The influence of pregnancy on blood selenium levels and glutathione peroxidase (GSH-Px) activity was studied in women. Whole blood and plasma selenium levels decreased whereas erythrocyte and plasma GSH-Px activities increased with the progress of pregnancy. The ratio of erythrocyte GSH-Px activity to whole blood selenium levels was 4- to 5-fold higher in rats and sheep than in primates (humans and monkeys), suggesting that more selenium is associated with GSH-Px in erythrocytes from rats and sheep than from primates. In assays of blood with low GSH-Px activity such as that from humans or selenium-deficient animals, a component of the erythrocyte other than GSH-Px was found to contribute more to the peroxidase activity. Evidence was obtained to indicate that t-butyl hydroperoxide is a better substrate than hydrogen peroxide for the assay of low GSH-Px erythrocyte activity. The length of time that the blood was stored before assay was found to have an effect on erythrocyte GSH-Px activity, and the storage patterns may be dependent on the species of animal from which the blood is drawn.

Corticotropin releasing factor (CRF) activity in rat plasma.

The ACTH-releasing activity of hypothalamic extract and rat plasma was examined with the dispersed rat pituitary cell technique of Swallow and Sayer (8). Although both plasma and serum caused ACTH release which was dose-related, stress did not enhance the ACTH releasing activity. Furthermore, separation studies of plasma using ultrafiltration and gel separation suggest that the CRF activity in plasma is associated with molecules of a molecular weight greater than 15,000.

The amino acid sequence of the alpha chain of the major haemoglobin of the rat (Rattus norvegicus).

1. A partial amino acid sequence of the alpha chain from the rat (Wistar, Rattus norvegicus) major haemoglobin is reported. The soluble tryptic peptides prepared from aminoethylated alpha-globin were separated by peptide 'mapping'. Sequencing of the tryptic peptides was carried out by the dansyl-Edman method and by the overlapping of smaller peptide fragments derived from secondary enzymic digestion. The insoluble 'core' peptides were further digested with chymotrypsin, thermolysin and pepsin to give smaller soluble peptides for sequencing. The tryptic peptides were ordered on the basis of their homology with the corresponding peptides of human alpha chain. 2. The proposed sequence is compared with that obtained by using an automated sequencer [Garrick et al. (1975) Biochem. J. 149, 245-258]. The differences in sequence resulting from the two methods are discussed. 3. It is suggested that the externally situated cysteine (residue 13) is responsible for the observed inhibition of crystallization of rat haemoglobin at alkaline pH. 4. Detailed evidence for the sequence has been deposited as Supplementary Publication SUP 50047 (9 pages) at the British Library (Linding Division), Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from which copies can be obtained on the terms given in Biochem. J. (1975) 145, 5.

An evaluation of the blood glucose lowering effects of Eryngium foetidium

a single oral dose of the aqueous leaf extract of Eryngium foetidum L. was screened for its blood-sugar lowering action in three animal models: normoglycaemic rats, streptozotocin-induced diabetic rats and normal rats subjected to the oral glucose-tolerance test. High and low doses (351 mg/kg and 176 mg/kg respectively) of the Eryngium foetidum leaf extract were used. The effects were composed with those produced by 200 mg/kg of the extract of commercial Gymnema sylvestre leaf (positive control 1), 3mg/kg glibenclamide (positive control 2) and 15mg/kg distilled water (negative control). Single (acute) oral dose of E. foetidium leaf extract caused no significant reduction in the blood glucose levels of the three animal models. The effect was similiar to that produced by the glibenclamide (positive control 2). The intraperitoneal acute toxicity test result in mice indicated that the E. foetidium leaf extract up to a dose of 702 mg/kg was not toxic. Phytochemical screening showed that essential oils and saponins were present in this extract. The present study suggests that a single oral dose of the aqueous leaf extract of E. foetidum has no significant blood-sugar lowering activity in healthy and experimental diabetic rats (AU)