High-fat diet elicits sex-based differences in liver inflammatory cytokines and redox homeostasis.

Sex differences in metabolic dysfunction-associated steatotic liver disease (MASLD) have been reported. Oxidative stress and inflammation are involved in the progression of MASLD. Thus, we aimed to evaluate liver redox homeostasis and inflammation in male and female rats fed a high-fat diet (HFD). Male and female Wistar rats were divided into the following groups: standard chow diet (SCD) or HFD during 12 weeks. HFD groups of both sexes had higher hepatocyte injury, with no differences between the sexes. Portal space liver inflammation was higher in females-HFD compared to females-SCD, whereas no differences were observed in males. Lobular inflammation and overall liver inflammation were higher in HFD groups, regardless of sex. TNF-α, IL-6, and IL-1ß levels were higher in males-HFD compared to males-SCD, but no differences were observed in females. Catalase activity was higher in males compared to females, with no differences between the SCD and HFD groups of both sexes. Glutathione peroxidase activity was higher in females compared to males, with no differences between the SCD and HFD groups in both sexes. Lipid peroxidation was higher in female-SCD when compared to male-SCD, and in both male- and female-HFD compared to SCD groups. Furthermore, both cytoplasmic and nuclear NRF2 staining were lower in the HFD group compared to the SCD group in males. However, female-HFD exhibited reduced nuclear NRF2 staining compared to the female-SCD group. In conclusion, our study demonstrated that while both male and female rats developed metabolic dysfunction-associated steatohepatitis after 12 weeks of HFD, the alterations in inflammatory cytokines and redox balance were sexually dimorphic.

Evaluation of the relationship between nutritional status, levels of physical activity and physical strength in adolescents.

BACKGROUND & AIMS: The study aimed at evaluating the relationship among anthropometric measurements, levels of physical activity and physical fitness in schoolchildren. METHODS: A cross-sectional study was carried out in public schools, with 173 adolescents from 10 to 17 years of age. Socioeconomic data were obtained by a structured questionnaire. Anthropometric measures were assessed and classified according to Body Mass Index (BMI)/age and Waist Circumference (WC). The long version of the International Physical Activity Questionnaire (IPAQ) was used to assess the level of physical activity, while the physical fitness level was assessed using the Projeto Esporte Brasil (PROESP) test protocol. RESULTS: 72,3% of the adolescents were eutrophic and 24.3% were overweight and 22.6% were at high risk for cardiovascular disease, with no statistical difference between the sexes (p > 0.05). 53.8% were physically inactive, regardless of sex and nutritional status. 86.1% of the adolescents showed low physical fitness, more significantly for sit-and-reach andsquare tests of females. BMI was directly correlated with physical fitness in the assessment ofupper limb power and agility (medicine ball throw and square test) and indirectly with aerobic capacity and lower limb power (abdominal resistance, horizontal jump and general resistance). The opposite was observed in the correlation of endurance (abdominal and general) and medicine ball throw with WC. Also, there was a likely visceral obesity and consequent cardiovascular risk in females more than in males. CONCLUSIONS: These findings reinforce the connection between physical activity and the presence of overweight and obesity in adolescents and also the need to effectively intervene in this groupin order to ensure the prevention of chronic non-communicable diseases in adulthood.

Improvement of 2,2'-Azobis(2-Methylpropionamidine) Dihydrochloride-Induced Hepatic Redox Imbalance in Swiss Mice and HepG2 Cells by Rutin.

Redox imbalance can lead to irreversible damages to biological functions. In this context, rutin stands out for its antioxidant potential. The objective of this study was to evaluate the acute and chronic effect of rutin on the hepatic redox imbalance. The study was performed according to three different protocols. First, healthy male Swiss mice were divided into two groups: control and rutin, the second of which received chronic oral supplementation of rutin (10 mg/kg). The second involved evaluation of the generation of reactive oxygen species (ROS) by HepG2 cells, incubated or not with rutin (20 and 40 µg/mL) for 3 h. The final protocol involved assessment of the acute effect of rutin (10 mg/kg) in mice with oxidative stress induced by 2,2'-azobis(2-methylpropionamidine) dihydrochloride (ABAP). After the in vivo treatments, the livers were collected to analyze the oxidative damage by thiol, and the antioxidant defense by catalase, superoxide dismutase, and glutathione peroxidase. In the HepG2 cells, the following probes were employed to assess the ROS production: dichlorofluorescein, MitoSOX, dihydroethidium, and Amplex Red. Rutin administered chronically improved the antioxidant defense in healthy animals, and when administered acutely both inhibited the increased production of ROS in HepG2 cells and improved the redox imbalance parameters in mice with induced oxidative stress. This study suggests rutin as a protective agent for restoration of hepatic redox homeostasis in redox injury induced by ABAP in Swiss mice and HelpG2 cells.

Phytomodulatory proteins isolated from Calotropis procera latex promote glycemic control by improving hepatic mitochondrial function in HepG2 cells.

The medicinal uses of Calotropis procera are diverse, yet some of them are based on effects that still lack scientific support. Control of diabetes is one of them. Recently, latex proteins from C. procera latex (LP) have been shown to promote in vivo glycemic control by the inhibition of hepatic glucose production via AMP-activated protein kinase (AMPK). Glycemic control has been attributed to an isolated fraction of LP (CpPII), which is composed of cysteine peptidases (95%) and osmotin (5%) isoforms. Those proteins are extensively characterized in terms of chemistry, biochemistry and structural aspects. Furthermore, we evaluated some aspects of the mitochondrial function and cellular mechanisms involved in CpPII activity. The effect of CpPII on glycemic control was evaluated in fasting mice by glycemic curve and glucose and pyruvate tolerance tests. HepG2 cells was treated with CpPII, and cell viability, oxygen consumption, PPAR activity, production of lactate and reactive oxygen species, mitochondrial density and protein and gene expression were analyzed. CpPII reduced fasting glycemia, improved glucose tolerance and inhibited hepatic glucose production in control animals. Additionally, CpPII increased the consumption of ATP-linked oxygen and mitochondrial uncoupling, reduced lactate concentration, increased protein expression of mitochondrial complexes I, III and V, and activity of peroxisome-proliferator-responsive elements (PPRE), reduced the presence of reactive oxygen species (ROS) and increased mitochondrial density in HepG2 cells by activation of AMPK/PPAR. Our findings strongly support the medicinal use of the plant and suggest that CpPII is a potential therapy for prevention and/or treatment of type-2 diabetes. A common epitope sequence shared among the proteases and osmotin is possibly the responsible for the beneficial effects of CpPII.

Effects of consumption of acerola, cashew and guava by-products on adiposity and redox homeostasis of adipose tissue in obese rats.

BACKGROUND: Fruit by-products contain phytochemicals, fibers and other components that can improve the redox imbalance of obesity. OBJECTIVE: The objective was to evaluate the effects of consumption of by-products of acerola, cashew and guava on the adiposity and redox homeostasis of adipose tissue in obese rats. METHODS: The animals were separated into 5 groups, control (CTL), high fat (HF), HF supplemented with acerola (HFA), cashew (HFC) and guava (HFG). RESULTS: Thiol quantification, lipid profile, catalase (CAT) and glutathione peroxidase (GPX) test were performed. TGL and VLDL levels were increased in HF group, and the treated groups did not change the lipid profile. CAT activity was increased in HFA and HFG groups. HFA was able to reduce the weight of the subcutaneous cushion. CONCLUSION: Treatment with fruit by-products did not alter weight gain, energy efficiency and body weight. Thus, the by-products of acerola and guava can be used as a sustainable alternative in the treatment of obesity.

Toxic stress, health and nutrition among Brazilian children in shelters.

BACKGROUND: Living in a shelter is an adverse experience that generates toxic stress. This situation can cause the dysregulation of the hypothalamic-pituitary-adrenal axis and exert a negative impact on health.The aim of the present study was to determine the association between toxic stress and social, clinical and nutritional characteristics in children at welfare institutions in a city of northeastern of Brazil. METHODS: An analytical, cross-sectional study was conducted with male and female children up to 60 months of age who live in shelters. Hair cortisol was used for the assessment of stress (immunoassay). The anthropometric data collected were height for age, body mass index for age, arm circumference for age, and head circumference for age (expressed in z-scores). We also evaluated food intake using markers proposed by the Brazilian Dietary and Nutritional Vigilance Surveillance System as well as the occurrence of dental caries and anemia. RESULTS: Sixty-three children one to 60 months of age participated in the present study. Asthma was the most frequent disease (11.1%). The prevalence of short stature, anemia and dental caries in the sample was 22.2, 22.2 and 9.4%, respectively. Cortisol levels ranged from 0.93 pg/mg to 391.29 pg/mg (median: 6.17 pg/mg). Higher cortisol levels were found in children with illnesses (p = 0.012) and those who had been hospitalized after being admitted to the institutions (p = 0.001). CONCLUSIONS: The majority of children had unhealthy eating behavior. The cortisol concentrations found in the present study were suggestive of dysregulation of the hypothalamic-pituitary-adrenal axis. Hypercortisolism was associated with illness and hospitalization.

High-dose Nandrolone Decanoate induces oxidative stress and inflammation in retroperitoneal adipose tissue of male rats.

The non-therapeutic use of the androgenic anabolic steroid Nandrolone Decanoate is popular due to its effects on physical performance and body composition, especially for its lipolytic and anabolic effects associated. However, high doses of such drugs are often associated with a series of pathologies related to unbalanced redox homeostasis, which, in turn, can be linked to inflammation. The oxidative stress onset could deregulate the secretion of cytokines, evidencing a dysfunctional adipocyte. Thus, the aim of this study was to investigate the effect of supraphysiological doses of Nandrolone Decanoate on redox homeostasis of retroperitoneal fatpad of male rats and its relationship with cytokines-based inflammatory signaling. Hydrogen peroxide production was assessed in the retroperitoneal fat pad of adult male rats which received either 10 mg kg of Nandrolone Decanoate or only a vehicle. Also, catalase, superoxide dismutase and glutathione peroxidase activities were measured, together with total reduced thiols and protein carbonylation, as well as IL-1ß, TNF-α, and IL-6 local levels. High doses of Nandrolone Decanoate caused an increase in the hydrogen peroxide production, together with lower activities of the antioxidant enzymes and lower levels of total reduced thiol. There were also higher protein carbonylation and greater levels of IL-1ß, TNF-α, and IL-6 in the treated group compared to control group. Therefore, it was possible to verify that high doses of Nandrolone Decanoate cause oxidative stress and induce higher inflammatory signaling in retroperitoneal fat pad of male rats.

Dieta hiperlipídica induz danos hepáticos associados à dhgna sem desequilíbrio redox no fígado de camundongos Swiss / Hyperlipid diet induces NAFLD associated liver damage without redox imbalance in Swiss mice liver

A doença hepática gordurosa não alcoólica (DHGNA) é uma doença de prevalência crescente e fortemente associada ao desequilíbrio redox e à perda da homeostase glicêmica. O desenvolvimento de modelos in vivo que mimetizem características fisiopatológicas da mesma é essencial para a descoberta e o desenvolvimento de estratégias terapêuticas. Validar um modelo de DHGNA associado ao desequilíbrio redox e à intolerância à glicose, induzida por dieta hiperlipídica, de baixo custo e fácil reprodutibilidade. Dezesseis camundongos Swiss receberam dieta hiperlipidica, durante 10 semanas. Durante todo o experimento foram avaliados peso corporal, tolerância à glicose, marcadores do estresse oxidativo e morfologia hepática. A dieta hiperlipídica promoveu um aumento significativo dos níveis de triglicerídeos (p<0,05) e do peso do tecido hepático (p<0,01), além de severa inflamação, necrose e esteatose dos hepatócitos. Os animais que seguira a DH (dieta hiperlipídica) apresentaram ainda, uma menor tolerância à glicose (p <0,05), entretanto não apresentaram redução dos grupamentos tióis. O modelo de DHGNA, induzida por DH, foi associada a danos histológicos hepáticos e à intolerância à glicose, entretanto não apresentou desequilíbrio redox, sendo um modelo inviável para estudos com tratamentos com antioxidantes. O trabalho teve por objetivo validar um modelo de DHGNA induzido por dieta hiperlipídica associada a alterações na resposta glicêmica e sem modificações na homeostase redox em camundongos.(AU)

Non-alcoholic fatty liver disease (NAFLD) is an increasingly prevalent disease strongly associated with redox imbalance and loss of glycemic homeostasis. The development of in vivo models that mimics its pathophysiological characteristics is essential for the discovery and development of therapeutic strategies. To validate a NAFLD model associated with redox imbalance and low-fat diet-induced glucose intolerance and easily reproducible. Sixteen Swiss mice received a high fat diet for 10 weeks. Throughout the experiment, body weight, glucose tolerance, oxidative stress markers and liver morphology were evaluated. The hyperlipid diet promoted a significant increase in triglyceride levels (p<0.05) and liver tissue weight (p <0.01), in addition to severe inflammation, necrosis and steatosis of hepatocytes. The animals that followed the hyperlipidic diet (HD) still had a lower glucose tolerance (p<0.05), ), however it did not show reduction of thiol groups. The HD-induced NAFLD model was associated with liver histological damage and glucose intolerance, however it did not present redox imbalance, being an unfeasible model for studies with antioxidant treatments. The study aimed to validate a model of NAFLD induced by a highfat diet associated with changes in the glycemic response and without changes in redox homeostasis in mice.(AU)

Dieta hiperlipídica induz danos hepáticos associados à dhgna sem desequilíbrio redox no fígado de camundongos Swiss / Hyperlipid diet induces NAFLD associated liver damage without redox imbalance in Swiss mice liver

A doença hepática gordurosa não alcoólica (DHGNA) é uma doença de prevalência crescente e fortemente associada ao desequilíbrio redox e à perda da homeostase glicêmica. O desenvolvimento de modelos in vivo que mimetizem características fisiopatológicas da mesma é essencial para a descoberta e o desenvolvimento de estratégias terapêuticas. Validar um modelo de DHGNA associado ao desequilíbrio redox e à intolerância à glicose, induzida por dieta hiperlipídica, de baixo custo e fácil reprodutibilidade. Dezesseis camundongos Swiss receberam dieta hiperlipidica, durante 10 semanas. Durante todo o experimento foram avaliados peso corporal, tolerância à glicose, marcadores do estresse oxidativo e morfologia hepática. A dieta hiperlipídica promoveu um aumento significativo dos níveis de triglicerídeos (p<0,05) e do peso do tecido hepático (p<0,01), além de severa inflamação, necrose e esteatose dos hepatócitos. Os animais que seguira a DH (dieta hiperlipídica) apresentaram ainda, uma menor tolerância à glicose (p <0,05), entretanto não apresentaram redução dos grupamentos tióis. O modelo de DHGNA, induzida por DH, foi associada a danos histológicos hepáticos e à intolerância à glicose, entretanto não apresentou desequilíbrio redox, sendo um modelo inviável para estudos com tratamentos com antioxidantes. O trabalho teve por objetivo validar um modelo de DHGNA induzido por dieta hiperlipídica associada a alterações na resposta glicêmica e sem modificações na homeostase redox em camundongos.

Non-alcoholic fatty liver disease (NAFLD) is an increasingly prevalent disease strongly associated with redox imbalance and loss of glycemic homeostasis. The development of in vivo models that mimics its pathophysiological characteristics is essential for the discovery and development of therapeutic strategies. To validate a NAFLD model associated with redox imbalance and low-fat diet-induced glucose intolerance and easily reproducible. Sixteen Swiss mice received a high fat diet for 10 weeks. Throughout the experiment, body weight, glucose tolerance, oxidative stress markers and liver morphology were evaluated. The hyperlipid diet promoted a significant increase in triglyceride levels (p<0.05) and liver tissue weight (p <0.01), in addition to severe inflammation, necrosis and steatosis of hepatocytes. The animals that followed the hyperlipidic diet (HD) still had a lower glucose tolerance (p<0.05), ), however it did not show reduction of thiol groups. The HD-induced NAFLD model was associated with liver histological damage and glucose intolerance, however it did not present redox imbalance, being an unfeasible model for studies with antioxidant treatments. The study aimed to validate a model of NAFLD induced by a highfat diet associated with changes in the glycemic response and without changes in redox homeostasis in mice.

Sex differences in subcutaneous adipose tissue redox homeostasis and inflammation markers in control and high-fat diet fed rats.

The development of obesity-related metabolic disorders is more evident in male in comparison with female subjects, but the mechanisms are unknown. Several studies have shown that oxidative stress is involved in the pathophysiology of obesity, but the majority of these studies were performed with male animals. The aim of this study was to evaluate the sex-related differences in subcutaneous adipose tissue redox homeostasis and inflammation of rats chronically fed a high-fat diet. NADPH oxidase (NOX), glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase activities were evaluated in the subcutaneous adipose tissue (SC) of adult male and female rats fed either a standard chow (SCD) or a high-fat diet (HFD) for 11 weeks. NOX2 and NOX4 messenger RNA (mRNA) levels, total reduced thiols, interleukin (IL)-1ß, tumor necrosis factor α (TNF-α), and IL-6 were also determined. Higher antioxidant enzyme activities and total reduced thiol levels were detected in SC of control male compared with female rats. Chronic HFD administration increased NOX activity and NOX2 and NOX4 mRNA levels and decreased SOD and GPx activities only in male animals. IL-1ß, TNF-α, and IL-6 levels, as well as Adgre1, CD11b, and CD68 mRNA levels, were also higher in SC of males after HFD feeding. In SC of females, catalase activity was higher after HFD feeding. Taken together, our results show that redox homeostasis and inflammation of SC is sexually dimorphic. Furthermore, males show higher oxidative stress in SC after 11 weeks of HFD feeding owing to both increased reactive oxygen species (ROS) production through NOX2 and NOX4 and decreased ROS detoxification.

Phytomodulatory proteins promote inhibition of hepatic glucose production and favor glycemic control via the AMPK pathway.

Phytomodulatory proteins from the latex of the medicinal plant Calotropis procera has been shown to be implicated in many pharmacological properties. However there is no current information about their activity on glucose metabolism, although the latex is used in folk medicine for treating diabetes. In this study the phytomodulatory proteins (LP) from C. procera latex were assessed on glycemic homeostasis. Control animals received a single intravenous dose (5 mg/kg) of LP or saline solution (CTL). Four hours after treatment, the animals were euthanized and their livers were excised for analysis by western blot and RT-PCR AMP-activated protein kinase (AMPK), phosphoenolpyruvate carboxykinase (PEPCK) and tumor necrosis factor alpha (TNF-α). In vivo tests of intraperitoneal tolerance to insulin, glucose and pyruvate were also performed, and the effect of LP administration on fed glycemia was studied followed by blood analysis to determine serum insulin levels. Treatment with LP reduced glycemia two hours after glucose administration (LP: 87.2 ± 3.70 mg/dL versus CTL: 115.6 ± 8.73 mg/dL). However, there was no change in insulin secretion (CTL: 14.16 ± 0.68 mUI/mL and LP: 14.96 ± 0.55 mUI/mL). LP improved the insulin sensitivity, represented by a superior glucose decay constant during an insulin tolerance test (kITT) (4.17 ± 0.94%/min) compared to the CTL group (0.82 ± 0.72%/min), and also improved glucose tolerance at 30 min (105.2 ± 12.4 mg/dL versus 154.2 ± 18.51 mg/dL), while it decreased hepatic glucose production at 15 and 30 min (LP: 75.5 ± 9.31 and 52.5 ± 12.05 mg/dL compared to the CTL: 79.0 ± 3.02 and 84.5 ± 7.49 mg/dL). Furthermore, there was a significant inhibition of gene expression of PEPCK (LP: 0.66 ± 0.06 UA and CTL: 1.14 ± 0.22 UA) and an increase of phosphorylated AMPK (LP: 1.342 ± 0.21 UA versus CTL: 0.402 ± 0.09 UA). These findings confirm the effect of LP on glycemic control and suggest LP may be useful in diabetes treatment. However, the pharmacological mechanism of LP in PEPCK modulation still needs more clarification.

Combined treatment with melatonin and insulin improves glycemic control, white adipose tissue metabolism and reproductive axis of diabetic male rats.

AIMS: Melatonin treatment has been reported to be capable of ameliorating metabolic diabetes-related abnormalities but also to cause hypogonadism in rats. We investigated whether the combined treatment with melatonin and insulin can improve insulin resistance and other metabolic disorders in rats with streptozotocin-induced diabetes during neonatal period and the repercussion of this treatment on the hypothalamic-pituitary-gonadal axis. MAIN METHODS: At the fourth week of age, diabetic animals started an 8-wk treatment with only melatonin (0.2 mg/kg body weight) added to drinking water at night or associated with insulin (NHP, 1.5 U/100 g/day) or only insulin. Animals were then euthanized, and the subcutaneous (SC), epididymal (EP), and retroperitoneal (RP) fat pads were excised, weighed and processed for adipocyte isolation for morphometric analysis as well as for measuring glucose uptake, oxidation, and incorporation of glucose into lipids. Hypothalamus was collected for gene expression and blood samples were collected for biochemical assays. KEY FINDINGS: The treatment with melatonin plus insulin (MI) was capable of maintaining glycemic control. In epididymal (EP) and subcutaneous (SC) adipocytes, the melatonin plus insulin (MI) treatment group recovered the insulin responsiveness. In the hypothalamus, melatonin treatment alone promoted a significant reduction in kisspeptin-1, neurokinin B and androgen receptor mRNA levels, in relation to control group. SIGNIFICANCE: Combined treatment with melatonin and insulin promoted a better glycemic control, improving insulin sensitivity in white adipose tissue (WAT). Indeed, melatonin treatment reduced hypothalamic genes related to reproductive function.

n5-STZ Diabetic Model Develops Alterations in Sciatic Nerve and Dorsal Root Ganglia Neurons of Wistar Rats.

One experimental model of diabetes mellitus (DM) similar to type II DM, called n5-STZ, is obtained by a single injection (via i.p.) of streptozotocin (STZ) in the 5th day of life of newborn rats. The present investigation aimed to characterize alterations in excitability of rat peripheral neurons in n5-STZ model. n5-STZ DM was induced, and electrophysiological evaluation was done at 12th week of rat life. Rats developed glucose intolerance, sensory alteration, and hyperglycemia or near-normoglycemia (21.2 ± 1.6 and 7.4 ± 0.4 mmol/L). In near-normoglycemia group the significant electrophysiological alteration observed was decreased in amplitude of 2nd wave (2nd component, conduction velocity: 48.8 m/s) of compound action potential (CAP) of sciatic nerve. For hyperglycemic rats, decreased excitability, amplitude, and conduction velocity of 2nd CAP component of sciatic nerve were found; a depolarization of resting potential (4-5 mV) and reduction in maximum ascendant and descendant inclinations of action potential were found in DRG neurons but no alteration on Na(+) current (INa(+) ). Thus, n5-STZ rats develop alterations in excitability which were related to glycemic levels but were not likely attributable to changes on INa(+) . Our data confirm that n5-STZ model is a useful model to study type II DM.