Vitamin D restriction enhances periovarian adipose tissue inflammation in a model of menopause.

Background: Menopause and deficiency in vitamin D (VD) are two health problems usually associated with aging women.Objective: We aimed to study inflammation in visceral adipose tissue when bilateral ovariectomy is combined with dietary restriction in VD.Methods: We studied 60 female C57BL/6 mice 3 months of age. Half of the animals had bilateral ovariectomy (Ovx group, n = 30) and half a sham procedure (Control [C] group, n = 30), and half of each Ovx or C group were fed a standard diet containing VD or a diet restricted in VD (D-) for an additional 12 weeks. Therefore, four groups were formed (n = 15 each group): C, C(D-), Ovx, and Ovx(D-). After sacrifice, the periovarian adipose tissue (PAT) was investigated.Results: In PAT, we observed different levels of hypertrophied adipocytes, enhanced proinflammatory cytokines, activation of inflammatory markers, and components of the extracellular signal-regulated kinase. The most affected PAT was seen in the Ovx(D-) group, followed by the Ovx group, the C(D-) group, and the C group (the least altered).Conclusion: The results demonstrate that ovariectomy and dietary restriction of VD are inducers of adverse effects on mouse visceral adipose tissue. When combined, these insults might enhance PAT inflammation.

Beneficial effects of maternal swimming during pregnancy on offspring metabolism when the father is obese.

We aimed to evaluate the impact of maternal exercise training on the offspring metabolism and body size caused by father obesity. C57BL/6 male 4-week-old mice were fed a high-fat diet (HF father) or control diet (C father), while equal age female mice were fed only a C diet and were separated into two groups: trained (T mother) and non-trained (NT mother), and at 12 weeks of age mice were mated. A continuous swimming protocol was applied for 10 weeks (before and during gestation), and offspring were followed since weaning until sacrifice (at 12 weeks of age). HF father, compared to C father, showed obesity, elevated total cholesterol (TC) and triglycerides (TG), and glucose intolerance. Both sexes HF/NT offspring showed hyperglycemia, glucose intolerance and high levels of TC and TG, without obesity. However, HF/T offspring showed data close to C/NT, demonstrating the beneficial effect of maternal exercise in the offspring of obese fathers.

Liver metabolism in adult male mice offspring: consequences of a maternal, paternal or both maternal and paternal high-fructose diet.

The study aimed to evaluate the consequences of the consumption of a high-fructose diet (HFR; fructose was responsible for 45% of the energy from carbohydrates) by the mother, the father, or both on C57BL/6 adult male offspring. Non-consanguineous parents received the diet (HFR or control, C) from 8 weeks before mating until weaning (n=10 fathers and n=10 mothers on each diet). After weaning, only the C diet was offered to offspring. The groups were formed by one male randomly taken from each litter. The offspring groups were identified according to the mother's diet (the first letter), then the father's diet (the second letter), that is, C/C, C/HFR, HFR/C, HFR/HFR (n=10 per group). The parents exhibited the following characteristics: compared with those of the C group, the HFR parents had higher blood pressure (BP), enlarged liver, increased hepatic triacylglycerol content, hypercholesterolemia, hypertriglyceridemia, high plasma leptin and low adiponectin. The offspring exhibited the following characteristics: compared with the C/C group, the HFR/HFR group had high BP. The C/HFR, HFR/C and HFR/HFR showed elevated uric acid and leptin levels and diminished adiponectin. The HFR/HFR group showed liver inflammation (increased NFκB, SOCS3, JNK, TNF-α, IL1-ß and IL6 levels). Likewise, SREBP-1c and FAS were upregulated. In conclusion, the consumption of a HFR by the mother and/or father is associated with adverse effects on liver metabolism in adult male offspring. When both mother and father are fed a HFR, the adverse effects on the offspring are more severe.

High-intensity interval training (swimming) significantly improves the adverse metabolism and comorbidities in diet-induced obese mice.

AIM: Controlling obesity and other comorbidities in the population is a challenge in modern society. High-intensity interval training (HIIT) combines short periods of high-intensity exercise with long recovery periods or a low-intensity exercise. The aim was to assess the impact of HIIT in the context of diet-induced obesity in the animal model. METHODS: C57BL/6 mice were fed one of the two diets: standard chow (Lean group - LE) or a high--fat diet (Obese group - OB). After twelve weeks, the animals were divided into non-trained groups (LE--NT and OB-NT) and trained groups (LE-T and OB--T), and began an exercise protocol. For biochemical analysis of inflammatory and lipid profile, we used a colorimetric enzymatic method and an automatic spectrophotometer. One-way ANOVA was used for statistical analysis of the experimental groups with Holm-Sidak pos hoc test. Two-way ANOVA analyzed the interactions between diet and HIIT protocol. RESULTS: HIIT leads to significant reductions in body mass, blood glucose, glucose tolerance and hepatic lipid profile in T-groups compared to NT-groups. HIIT was able to reduce plasma levels of inflammatory cytokines. Additionally, HIIT improves the insulin immunodensity in the islets, reduces the adiposity and the hepatic steatosis in the T-groups. HIIT improves beta--oxidation and peroxisome proliferator--activated receptor (PPAR)-alpha and reduces lipogenesis and PPAR--gamma levels in the liver. In skeletal muscle, HIIT improves PPAR--alpha and glucose transporter-4 and reduces PPAR--gamma levels. CONCLUSION: HIIT leads to attenuate the adverse effects caused by a chronic ingestion of a high-fat diet.

Developmental origins of health and disease: experimental and human evidence of fetal programming for metabolic syndrome.

The concept of developmental origins of health and disease has been defined as the process through which the environment encountered before birth, or in infancy, shapes the long-term control of tissue physiology and homeostasis. The evidence for programming derives from a large number of experimental and epidemiological observations. Several nutritional interventions during diverse phases of pregnancy and lactation in rodents are associated with fetal and neonatal programming for metabolic syndrome. In this paper, recent experimental models and human epidemiological studies providing evidence for the fetal programming associated with the development of metabolic syndrome and related diseases are revisited.

High fat diets modulate nitric oxide biosynthesis and antioxidant defence in red blood cells from C57BL/6 mice.

The consumption of a high fat (HF) diet is considered a risk factor for the development of obesity. On the other hand, a monounsaturated HF diet has beneficial cardiometabolic effects. Since nitric oxide (NO) modulates vascular homeostasis, we investigate whether HF diets that vary in fatty acid composition have a different effect on theL-arginine-NO pathway and oxidative stress in C57BL/6 mice red blood cells (RBC). The olive oil diet induced an activation of L-arginine transport compared to other diets. NO synthase (NOS) activity was increased in all unsaturated HF diets (olive, sunflower and canola oils). Moreover, the expression of endothelial NOS (eNOS) and inducible NOS (iNOS) was increased in the olive oil group. In contrast, NOS activity from the lard group was decreased associated with diminished l-arginine transport. Olive oil also induced superoxide dismutase activation. Inhibition of the L-arginine-NO pathway in the lard group could contribute to cardiovascular diseases, while unsaturated HF diets may have a protector effect via enhanced NO bioavailability.

Beneficial effects of exercise training (treadmill) on insulin resistance and nonalcoholic fatty liver disease in high-fat fed C57BL/6 mice.

C57BL/6 mice develop signs and symptoms comparable, in part, to the human metabolic syndrome. The objective of the present study was to evaluate the effects of exercise training on carbohydrate metabolism, lipid profile, visceral adiposity, pancreatic islet alterations, and nonalcoholic fatty liver disease in C57BL/6 mice. Animals were fed one of two diets during an 8-week period: standard (SC, N = 12) or very high-fat (HF, N = 24) chow. An exercise training protocol (treadmill) was then established and mice were divided into SC and HF sedentary (SC-Sed, HF-Sed), exercised groups (SC-Ex, HF-Ex), or switched from HF to SC (HF/SC-Sed and HF/SC-Ex). HF/HF-Sed mice had the greatest body mass (65% more than SC/SC-Sed; P < 0.0001), and exercise reduced it by 23% (P < 0.0001). Hepatic enzymes ALP (+80%), ALT (+100%) and AST (+70%) were higher in HF/HF mice than in matched SC/SC. Plasma insulin was higher in both the HF/HF-Sed and HF/SC-Sed groups than in the matched exercised groups (+85%; P < 0.001). Pancreatic islets, adipocytes and liver structure were greatly affected by HF, ultimately resulting in islet beta-cell hypertrophy and severe liver steatosis. The HF group had larger islets than the SC/SC group (+220%; P < 0.0001), and exercise significantly reduced liver steatosis and islet size in HF. Exercise attenuated all the changes due to HF, and the effects were more pronounced in exercised mice switched from an HF to an SC diet. Exercise improved the lipid profile by reducing body weight gain, visceral adiposity, insulin resistance, islet alterations, and fatty liver, contributing to obesity and steatohepatitis control.

Beneficial effects of exercise training (treadmill) on insulin resistance and nonalcoholic fatty liver disease in high-fat fed C57BL/6 mice

C57BL/6 mice develop signs and symptoms comparable, in part, to the human metabolic syndrome. The objective of the present study was to evaluate the effects of exercise training on carbohydrate metabolism, lipid profile, visceral adiposity, pancreatic islet alterations, and nonalcoholic fatty liver disease in C57BL/6 mice. Animals were fed one of two diets during an 8-week period: standard (SC, N = 12) or very high-fat (HF, N = 24) chow. An exercise training protocol (treadmill) was then established and mice were divided into SC and HF sedentary (SC-Sed, HF-Sed), exercised groups (SC-Ex, HF-Ex), or switched from HF to SC (HF/SC-Sed and HF/SC-Ex). HF/HF-Sed mice had the greatest body mass (65 percent more than SC/SC-Sed; P < 0.0001), and exercise reduced it by 23 percent (P < 0.0001). Hepatic enzymes ALP (+80 percent), ALT (+100 percent) and AST (+70 percent) were higher in HF/HF mice than in matched SC/SC. Plasma insulin was higher in both the HF/HF-Sed and HF/SC-Sed groups than in the matched exercised groups (+85 percent; P < 0.001). Pancreatic islets, adipocytes and liver structure were greatly affected by HF, ultimately resulting in islet â-cell hypertrophy and severe liver steatosis. The HF group had larger islets than the SC/SC group (+220 percent; P < 0.0001), and exercise significantly reduced liver steatosis and islet size in HF. Exercise attenuated all the changes due to HF, and the effects were more pronounced in exercised mice switched from an HF to an SC diet. Exercise improved the lipid profile by reducing body weight gain, visceral adiposity, insulin resistance, islet alterations, and fatty liver, contributing to obesity and steatohepatitis control.

Different edible oil beneficial effects (canola oil, fish oil, palm oil, olive oil, and soybean oil) on spontaneously hypertensive rat glomerular enlargement and glomeruli number.

We have tested the different edible oil effects on the blood pressure (BP) control and the following glomerular protection. Six groups of 12-week-old male spontaneously hypertensive rats (SHR) (n = 5), have received different edible oils (fish, canola, palm, olive, and soybean) or a placebo by gavage for 13 weeks. Renal cortex was analyzed through light microscopy and stereology. Usual BP increase, glomerulosclerosis, glomerular enlargement, and glomeruli loss in SHR has been prevented (fish, canola and palm oils) or attenuated (olive and soybean oils) by these oil long-term administration. The most favorable effect has been seen in the fish oil administration (source of n-3 polyunsaturated fatty acids, PUFA, eicosapentaenoic and docosahexaenoic acids), followed by both canola and palm oils (source of n-3 PUFA plus n-9 monounsaturated, MUFA, and saturated fatty acid, respectively), and finally both olive and soybean oils (source of n-9 MUFA and n-6 PUFA, respectively).

Blood pressure, ventricular volume and number of cardiomyocyte nuclei in rats fed for 12 months on diets differing in fat composition.

Blood pressure (BP), body mass (BM), ventricular volume (V[vent]) and the number of ventricular cardiomyocyte nuclei (N[vcn]) were analysed in rats fed different dietary fats. A total of 20 Wistar male rats were studied from 21 days old to 12 months of age and divided in the groups: soybean oil (S), canola oil (CA), lard and egg yolk (LE) and canola oil+lard and egg yolk (CA+LE). The diets had the same basal diet that included cornstarch, casein, maize, egg white and mineral and vitamin mixtures. At the moment of the sacrifice, the LE group had the greatest BP and V[vent] which was significantly higher than the other groups, and the S group had the greatest BM. The myocardial structure was apparently normal in the S and CA groups while it showed areas of diffuse interstitial fibrosis and hypertrophied cardiomyocytes in the LE group, and intramyocardial coronaries with thick tunica media and little interstitial fibrosis in CA+LE group. The N[vcn] was significantly higher in the CA group and it was lower in the LE group. These results suggest that the different dietary fats affect the myocardial structure, and the canola oil diet reduces the cardiomyocyte loss in the old rats.

Myocardial stereological adaptations in wistar rats fed with different high-fat diets during 18 months.

This study has the purpose of investigating the influence of different high-fat experimental diets on myocardial structure in rats. Twenty-seven male rats were fed from 21 d old (postnatal age) until 18 mo old with one of the following supplemented diets: soybean oil (S) (n= 6), canola oil (CA) (n= 8), or lard and egg yolk (LE) (n= 6) or canola oil+ lard and egg yolk (CA+LE) (n=7). The blood pressure (BP) was measured, and after the sacrifice the cardiac biometry and the myocardial stereology were determined: cross-sectional area of cardiomyocyte (A), volume density (Vv), surface density (Sv), and length density (Lv) in relation to the cardiomyocytes (cm), connective tissue (ct), and blood vessels (v). The CA group rats had lower BP, A[cm], and Vv[ct]; they had greater Vv[cm], Sv[cm], Vv[v], Lv[v], and Sv[v] than the other groups. The S rats had intermediary values for the myocardium and blood vessel parameters between the CA and LE group rats. These results support the notion that the long-term use of canola oil in the diet is better to preserve the myocardium structure, including microvascularization, than soybean oil or lard and egg yolk.

Rupture of the ulnar collateral ligament of the metacarpophalangeal joint of the index finger.

We report a case of chronic instability due to rupture of the ulnar collateral ligament of the metacarpophalangeal joint of the index finger of the left hand. Because of persistent instability after the initial conservative treatment, surgical treatment was required.

Effect of different high-fat diets on the myocardium stereology and blood pressure in rats.

The effect of three high-fat diets containing 29% canola oil (CA), lard plus egg yolk (LE) or canola oil, lard and egg yolk (CA+LE) in male Wistar rats was investigated over a period of 6 months. We analyzed the myocardium, composed of cardiomyocytes and interstitium, which is made up of connective tissue and blood vessels. Volume density of cardiomyocyte (Vv[m]), volume density of blood vessels (Vv[v]), and volume density of connective tissue (Vv[ct]) were the stereological parameters determined. The rats of the LE group had a significantly higher heart mass/body mass ratio than those of the CA group. The blood pressure of the LE group was significantly higher than that of the other groups. In the CA group, the Vv[m] was significantly higher and the Vv[ct] was significantly lower than in the other groups. The myocardium of both the LE and CA+LE groups showed a significant reduction of Vv[m] and a compensatory increase of the Vv[ct]. These findings were less pronounced in the CA+LE group, in which the Vv[v] was found to be significantly higher than in the CA group. Comparing three high-fat diets, the data suggest that the diet canola oil had a major beneficial effect, preserving the myocardial structure and the blood pressure in rats.

Numerical density of cardiac myocytes in aged rats fed a cholesterol-rich diet and a canola oil diet (n-3 fatty acid rich).

We studied the myocardium of 45 aged rats fed from 21 days after birth until 15 months of age with a standard rat diet a cholesterol-rich diet (CHO) or canola oil (O). We analysed the cardiac weight (CW) and, using unbiased stereological estimates, studied isotropic, uniform, random sections of the free left ventricular wall to determine the numerical density of the myocytes (NV[myocyte]). The CW was not statistically different between groups A and CHO: it was smallest in animals in group O (21.2% smaller in group O than in group A and 15.3% smaller in group O than in group CHO). NV[myocyte] was statistically different in all three groups and was greatest in animals in group O. By comparison with rats in group A, group CHO rats had an NV[myocyte] than was 51.3% smaller and group O, 33.3% greater. Aged rats fed with canola oil diet have a well-vascularized myocardium, which is probably associated with preservation of NV[myocyte] in the myocardium of these animals.

Stereology of the myocardium and blood biochemistry in aged rats fed with a cholesterol-rich and canola oil diet (n-3 fatty acid rich).

The myocardial changes brought about by canola oil (n-3 fatty acid rich) and hyperlipidic diets were studied in 45 rats. Three groups each consisting of 15 animals was separated into (A) which receiving a normal balanced diet; and in groups (CHO) and (O) the animals receiving hyperlipidic and canola oil diet, respectively. These diets were fed to the animals from 21 days until 15 months old, then a blood analysis was performed, after which they were sacrificed and the hearts taken for light microscopic studies. The total lipids serum was extracted and the low density lipoproteins (LDL-C and VLDL-C) and chylomicron fractions were determined as well as the cholesterol concentration in the high density lipoprotein fraction (HDL-C). The myocardium was composed of myocytes and cardiac interstitium, which is made up of connective tissue and blood vessels. The following stereological parameters were determined: a) from myocyte: volume density of myocyte, total volume of myocytes surface density of myocyte, total surface of myocyte and cross sectional area of myocyte; b) from blood vessels: volume density of blood vessels, total volume of blood vessels, length density of blood vessels, surface density of blood vessels, total surface of blood vessels and cross sectional area of vessels; c) from connective tissue: volume density of connective tissue and total volume of connective tissue. The differences were tested by the analysis of variance and Tukey test. The Mantel-Haenezel test analyzed the survival curve test comparing the different groups. Many stereological parameters had significant differences: cardiac weight, thickness of the right and left ventricular wall, aorta and pulmonary artery inner diameters. HDL-C, LDL-C, volume density of myocyte, total surface of myocyte, surface density of myocyte, total surface of myocyte, total volume of blood vessel, length density of blood vessels, surface density of blood vessels, total surface of blood vessels, volume density of connective tissue, total volume of connective tissue. Differences in survival curves were significant between groups CHO x A and CHO x O (p < 0.05) but not between groups A x O (p = 0.48). For the cardiac weight, the smallest values were found in group O. The aorta and artery pulmonary internal diameters were smaller in group CHO. The HDL-C serum was about 40% greater in group O. The LDL-C serum was more than 80% less in the same group. The average of volume density of myocyte was less in group CHO, while the average of volume density of connective tissue was greater in group CHO in comparison to groups A and O. The length density of blood vessels was greater in group O than in groups A and CHO. The surface density of myocyte and surface density of blood vessels were smaller in group CHO and greater in group A. The total surface of myocyte and total surface of blood vessels were greater in group CHO and smaller in group O. Differences were significant between groups A x CHO. The total volume of myocytes was greater in group A, while the total volume of connective tissue was greater in group CHO. The cross sectional area of myocyte and cross sectional area of vessels were greater in group CHO and smaller in group O suggesting that the canola oil diet (n-3 fatty acid rich) preserves the myocardium more than the standard and cholesterol-rich diets.

[Stereology of the myocardium in young and aged rats]. / Estereologia do miocárdio de ratos jovens e idosos.

PURPOSE: To determine the myocardial quantitative changes comparing young and aged animals by using the stereology. METHODS: Thirty rat hearts were studied (15 rats aged 3 months and 15 other rats aging 15 months). The hearts were removed, weighed, fixed in 10% buffered formaldehyde solution, embedded in paraffin, cut in 7 microns thick slices and stained with HE and picro-sirius stains. In each group, we counted 15 random microscopic fields. The following parameters were studied: Vv (myocyte) and Vv(interstitium)(%) (the volume densities of the cardiac myocyte and interstitium, determined by the point-counting method), and Nv(myocyte) (1/mm3) (the numerical density of the cardiac myocytes, determined with the disector method). The total number of myocytes (N[myocyte]) and the mean volume of the myocytes (V[myocytes]) were also determined. The differences were tested by the Mann-Whitney test. RESULTS: Cardiac weight increased from 1.1 to 1.7 g, the Vv(myocyte) decreased from 76.7 to 72.2%, the Vv(interstitium) increased from 23.3 to 27.8%. The Nv(myocyte) and the N(myocyte) decreased from 14.76 x 10(4) to 6.19 x 10(4)/mm3 and 15.64 x 10(4) to 10.72 x 10(4) myocytes, respectively. Simultaneously, the V(myocyte) increased from 5.42 x 10(3) to 13.26 x 10(3) mm3. These differences were statistically significant (p < 0.05). CONCLUSION: Myocardial changes, comparing young rats with aged ones suggest loss of myocytes (increased apoptosis?) with simultaneous myocyte hypertrophy.

Estereologia do miocárdio de ratos jovens e idosos / Stereology of the myocytes in young and aged rats

OBJETIVO - Comparar as diferenças quantitativas na composiçäo e estrutura do miocárdio de ratos jovens e idosos utilizando a estereologia. MÉTODOS - Foram estudados 20 ratos machos da raça Winstar (15 animais jovens de 3 meses de idade e 15 animais idosos de 15 meses). Os coraçöes foram retirados, pesados, perfundidos com soluçäo de Bouin e posteriormente fixados em formol tamponado a 10 'por cento' por 24h, processados por técnica histológica, incluídos em parafina, seccionados e corados pelo HE e picro-sirius. Foram analisados 15 campos aleatórios por grupo e foram determinados os seguintes parâmetros estereológicos: densidade de volume do miócito (Vv[miócito]) e densidade de volume do interstício cardíaco (Vv[interstício]) e densidade numérica dos miócitos (Nv[miócito]) (1/mm 'ao cubo'), estimados pelo método disector. O número total de miócitos (N[miócitos]) e a média do volume do miócito (Vol[miócito]) dos coraçöes dos dois grupos também foram determinados. As diferenças estatísticas entre os animais jovens e idosos foram testadas pelo teste näo paramétrico de Mann-Whitney. RESULTADOS - Comparando os animais dos grupos jovem e idoso temos, respectivamente, os seguintes dados: o peso cardíaco aumentou de 1,1 para 1,7g, o Vv(miócito) diminuiu de 76,7 para 72,2 'por cento' Vv(interstício) aumentou de 23,3 para 27,8 'por cento', enquanto o Nv(miócito) diminuiu de 14,76 x 10 'à quarta' para 6,19 x 10'à quarta'/mm 'ao cubo'. O Vol (miócito) aumentou de 5,42 1x 10 'ao cubo' para 13,26 x 10 'ao cubo' e o N(miócito) diminiuiude 15,64 x 10 'à quarta' para 10,72 x 10 ' à quarta' miócitos. Estas diferenças foram estatisticamente significativas (p<0,05). CONCLUSÄO - Os resultados obtidos mostram que no envelhecimento ocorre perda de miócitos cardíacos e hipertrofia das células remanescentes.

[Morphological and biochemical comparison among aged rats fed with hyperlipidic and canola oil diet]. / Comparação morfológica e bioquímica entre ratos envelhecidos alimentados com dieta hiperlipídica e com óleo de canola.

PURPOSE: To observe cardiovascular and metabolic changes due to canola oil diet (rich in n-3 fatty acid). METHODS: Forty five rats were followed during 15 months separated in three groups. In group aged (A), they received a regular diet. In group H, animals received a hyperlipidic diet and in group O they received a canola oil diet. The rats were fed for 15 months after weaning. At the end of the experience a blood analysis was performed and heart and aorta were analyzed. The total lipids were extracted and the LDL, VLDL and HDL were determined. RESULTS: Total cholesterol and triglyceride levels were not different among groups. Differences were found in body and cardiac weight, the thickness of the right and left ventricular wall, aorta and pulmonary diameters, HDL and LDL (p < 0.05). Smallest values of the cardiac weight and thickness of the ventricular walls were found in group O. The aorta and pulmonary internal diameters were smaller in group H. The HDL was 40% greater in group O than in groups A and H while the LDL was more than 80% lower in group O than in groups A and H. Differences in survival curves between groups H x A and between H x O were significant (p < 0.05) but not between groups A x O (p = 0.48). CONCLUSION: Aged rats fed with canola oil diet (rich in n-3 fatty acid) presented morphological cardiovascular and metabolic changes less important in magnitude than old animals and, mainly, the same age animals under hyperlipidic diet.