β-Glucan administration to diabetic rats reestablishes redox balance and stimulates cellular pro-survival mechanisms

The mechanisms underlying the beneficial effect observed in diabetic rats after treatment with a commercially available β-glucan-enriched extract (BGEE) were examined. Multiple low-dose streptozotocin (STZ) diabetes was used (40 mg STZ/kg) as a model for type 1 diabetes. BGEE was administered daily (80 mg/kg) for 4 weeks, starting from the last day of the STZ treatment. In vitro and in vivo experiments suggest that significant free radical scavenging and antioxidant activities of BGEE were responsible for a systemic adjustment of the redox disturbance and reduction of DNA damage in the liver and kidney of diabetic rats. BGEE-treated diabetic rats also displayed increased Akt kinase activity and decreased pro-caspase-3 degradation, implying that BGEE mediates its beneficial effects through activation of the cellular pro-survival pathway. We conclude that β-glucan administration under diabetic conditions promotes a systemic improvement that can be expected to increase the organism’s resistance to the onset of diabetic complications.     

β-Casomorphin-7 cause decreasing in oxidative stress and inhibiting NF-κB-iNOS-NO signal pathway in pancreas of diabetes rats

β-Casomorphin-7 (β-CM-7) is a milk biological active peptide. The present study is aimed to investigate the protective effects of β-CM-7, against oxidative stress in pancreas of streptozotocin-induced diabetic rats by assaying malondialdehyde (MDA), nitric oxide (NO) level, the activity of enzymatic antioxidants such as superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-Px), and NF-κB, inducible nitric oxide synthase (iNOS) gene expression. A significant increase in the level of oxidative stress was observed in pancreas of the diabetic rats when compared to control rats. After 15 d oral administration of β-CM-7 (7.5 × 10(-8) mol/d), the pancreas MDA level was markedly reduced. Oral administration of β-CM-7 to diabetic rats showed an obviously increase in the activity of catalase in pancreas, oral administration of β-CM-7 to the diabetic group of rats also showed a reduction of NF-κB and iNOS gene expression in pancreas. The elevated pancreas NO level was markedly reduced by the oral administration of β-CM-7. Thus, the results of the present study suggest that β-CM-7 may cause protective effects such as pronounced decreasing in oxidative stress and inhibiting NF-κB-iNOS-NO signal pathway in pancreas of diabetes rats.     

Curcumin attenuates diabetic nephropathy by inhibiting PKC-α and PKC-β1 activity in streptozotocin-induced type I diabetic rats

Scope : We hypothesized that curcumin, a potent anti‐oxidant, might be beneficial in ameliorating the development of diabetic nephropathy through inhibition of PKC‐α and PKC‐β1 activity‐ERK1/2 pathway. Methods and results : Diabetes was induced by a single intraperitoneal injection of streptozotocin (STZ) (55 mg/kg) in rats. Three weeks after STZ injection, rats were divided into three groups, namely, normal, diabetic and diabetic treated with curcumin at 100 mg/kg/day, p.o., for 8 wk. At 11 wk after STZ injection, diabetic rats exhibited renal dysfunction, as evidenced by reduced creatinine clearance, increased blood urea nitrogen (BUN) and proteinuria, marked increases in lipid peroxidation, NOX4 and p67phox and decrease in anti‐oxidant enzyme. All of these abnormalities were significantly reversed by curcumin. Furthermore, the high‐glucose‐induced PKC‐α and PKC‐β1 activities and phosphorylated ERK1/2 was significantly diminished by curcumin. Curcumin also attenuated the expression of TGF‐β1, CTGF, osteopontin, p300 and ECM proteins such as fibronectin and type IV collagen. The high‐glucose‐induced expression of VEGF and its receptor VEGF receptor II (flk‐1) was also ameliorated by curcumin. Conclusion : These results prove that curcumin produces dual blockade of both PKC‐α and PKC‐β1 activities, which suggests that curcumin is a potential adjuvant therapy for the prevention and treatment of diabetic nephropathy.     

Adenosine 5′-monophosphate-activated protein kinase ameliorates bovine adipocyte oxidative stress by inducing antioxidant responses and autophagy

Adipose tissue concentration of reactive oxygen species (ROS) increases in dairy cows with ketosis, suggesting that the tissue experiences oxidative stress. Autophagy, an adaptive response to cellular stress, has been shown to promote survival and plays a critical role in antioxidant responses. Dysregulation of adenosine 5′-monophosphate-activated protein kinase (AMPK) is closely related to antioxidant responses and autophagy of adipocytes in animal models of metabolic disorders, but its role in bovine adipose tissue during periods of stress is unknown. We hypothesized that AMPK may play important roles in the regulation of oxidative stress in adipose tissue of ketotic cows. Specific objectives were to evaluate autophagy status and AMPK activity in adipose tissue of ketotic cows, and their link with oxidative stress in isolated bovine adipocytes. Selection of 15 healthy and 15 clinically ketotic Holstein cows at 17 (±4) d postpartum was performed after a thorough veterinary evaluation for clinical symptoms and also based on serum β-hydroxybutyrate concentrations before collection of subcutaneous adipose tissue samples. Primary cultures of bovine adipocytes isolated from the harvested adipose tissue were stimulated with varying concentrations of H₂O₂ (0, 50, 100, 200, or 400 μM) for 2 h. In another experiment, adipocytes were cultured with the AMPK activator A769662 or adenovirus-containing small interfering RNA (ad-AMPKα-siRNA) for 3 or 48 h, respectively, followed by H₂O₂ exposure (200 μM) for 2 h. Compared with healthy cows, clinical ketosis led to increased abundance of AMPK and nuclear factor erythroid-derived 2-like 2 (NFE2L2), but lower abundance of Kelch-like ECH-associated protein 1 (KEAP1) in adipose tissue. Abundance of the key proautophagy proteins Beclin1, sequestosome 1 (SQSTM1), autophagy-related gene 7 (ATG7), ATG5, and ratio of microtubule-associated protein light chain 3 (LC3) II to LC3I were greater in adipose tissue of ketotic cows. In bovine adipocytes, treatment with H₂O₂ induced accumulation of ROS and malondialdehyde (MDA), whereas H₂O₂ stimulation inhibited activities of the antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD). Addition of AMPK activator A769662 increased antioxidant response via activating NFE2L2 and its downstream targets heme oxygenase 1 (HMOX1), superoxide dismutase 1 (SOD1), catalase (CAT), and glutathione-S-transferase (GST) to improve H₂O₂-induced oxidative stress in adipocytes. Simultaneously, activation of AMPK increased abundance of Beclin1, SQSTM1, ATG7, ATG5, and ratio of LC3II to LC3I. In contrast, inhibition of AMPK downregulated abundance of NFE2L2, HMOX1, SOD1, CAT, Beclin1, SQSTM1, ATG7, ATG5, and ratio of LC3II to LC3I, and further aggravated H₂O₂-induced oxidative stress. Overall, these data indicate that activation of AMPK, as an adaptive mechanism for acute metabolic regulation of adipose tissue homeostasis, can induce antioxidant responses and autophagy, and further reduce oxidative stress in bovine adipocytes.     

Amelioration of cadmium-induced oxidative stress, impairment in lipids and plasma lipoproteins by the combined treatment with quercetin and α-tocopherol in rats

Cadmium (Cd) exposure results in numerous pathological consequences including oxidative stress and dyslipidemia. The present study was designed to investigate the efficacy of combined treatment with quercetin (QE) and α-tocopherol (AT) against Cd-induced oxidative stress and alterations in lipids and lipoproteins in the plasma and liver of rats. Oral administration of Cd (5 mg/kg bw/d) for 4 wk has shown a significant (P < 0.05) increase in thiobarbituric acid reactive substances (TBARS), lipid hydro peroxides (LOOH), total cholesterol, low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C), free fatty acids (FFA), phospholipids (PL), triglycerides (TGs), and the activity of hydroxyl-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase) in plasma with a significant (P > 0.05) reduction in the levels of reduced glutathione (GSH), high density lipoprotein cholesterol (HDL-C), and the activity of lecithin cholesterol acyl transferase (LCAT) in plasma. In addition, the levels of hepatic thiobarbituric acid reactive substances (TBARS), LOOH, conjugated dienes (CD), protein carbonyls (PC), and the activity of HMG-CoA reductase, levels of cholesterol, FFA, and TGs were significantly (P > 0.05) increased and the level of PL is significantly (P > 0.05) decreased along with the decreased activity of LCAT in the liver of Cd-treated rats. Oral supplementation with QE (50 mg/kg bw/d) and AT (50 mg/kg bw/d) for 4 wk in Cd intoxicated rats significantly (P > 0.05) has reduced the plasma levels of TBARS, LOOH, GSH, cholesterol, FFA, TGs, VLDL-C, LDL-C, and the activity of HMG-CoA and significantly (P > 0.05) has increased the activity of LCAT and the plasma levels of HDL-C. The oral supplementation also significantly (P > 0.05) has reduced the hepatic oxidative stress markers, cholesterol, TGs, FFA, and significantly (P > 0.05) has increased the LCAT activity and the PL in liver. Our results indicate that the combined treatment with QE and AT has normalized all the previously mentioned biochemical parameters in Cd-intoxicated rats than the individual treatments. The combined treatment has provided remarkable protection against Cd-induced oxidative stress and alterations in lipid metabolism and, thereby, reduced the Cd-mediated cardiovascular diseases.     

Dietary flaxseed oil and fish oil modulates expression of antioxidant and inflammatory genes with alleviation of protein glycation status and inflammation in liver of streptozotocin–nicotinamide induced diabetic rats

Beneficial effects of dietary flaxseed oil or fish oil on streptozotocin–nicotinamide induced diabetic rats were investigated. Rats were divided into three diabetic and three non-diabetic groups and received control, flaxseed oil or fish oil diets (10% w/w). Both diets reduced blood glucose, TBARS and hepatic NO. The extent of glycation measured in terms of glycated albumin and hemoglobin was reduced significantly with both diets. Flaxseed oil diet up-regulated hepatic catalase (CAT) (activity and expression), superoxide dismutase (SOD) (activity and expression) and glutathione peroxidase (GPx) expression. Fish oil diet up-regulated hepatic CAT (activity and expression), paraoxonase-1 (PON-1) expression and down-regulated heme oxygenase-1 (HO-1) expression. Furthermore, both diets down-regulated the expression of hepatic inflammatory genes TNF-α, IL-6, MCP-1, INF-γ and NF-κB. These results were supported by histopathological observations which showed better tissue preservation in both the diets. Thus, both the diets proved to be beneficial in preventing tissue injury and alleviating diabetic insults in the livers of STZ–NIC diabetic rats.     

Bioactive compounds extracted from Gamtae (Ecklonia cava) by using enzymatic hydrolysis, a potent α-glucosidase and α-amylase inhibitor, alleviates postprandial hyperglycemia in diabetic mice

This study was designed to investigate whether the brown alga gamtae (Ecklonia cava) may inhibit α-glucosidase and α-amylase activities, and alleviate postprandial hyperglycemia in streptozotocin-induced diabetic mice. For that purpose, we prepared an enzymatic hydrolysate from gamtae (EHG) by using the carbohydrase, Celluclast. EHG evidenced prominent inhibitory effect against α-glucosidase and α-amylase. The IC₅₀ values of EHG against α-glucosidase and α-amylase were 0.62 and 0.59 mg/mL, respectively, which evidenced the higher activities than that of acarbose. EHG did not exert any cytotoxic effect in human umbilical vein endothelial cells (HUVECs) at various concentrations (from 0.25 to 2 mg/mL). The increase of postprandial blood glucose levels were significantly suppressed in the EHG administered group than those in the streptozotocin-induced diabetic or normal mice. Moreover, the area under curve (AUC) was significantly reduced via EHG administration (6,102 vs. 10,425 mg·min/dL) in the diabetic mice as well as it delays absorption of dietary carbohydrates. These result indicated that EHG might be a potent inhibitor for α-glucosidase and α-amylase.     

β-Hydroxybutyrate (BHB) and glucose concentrations in the blood of dairy calves as influenced by age,vaccination stress,weaning, and starter intake including evaluation of BHB and glucose markers of starter intake

The objective of this research was to determine how blood β-hydroxybutyrate (BHB) and glucose are affected by age, time of day, stress, weaning, forced intake restriction, and voluntary starter intake in calves between 1 and 9 wk of age, and to evaluate if either is an acceptable proxy for starter intake. Holstein calves were fed a 27% crude protein, 17% fat milk replacer at 660 g of dry matter daily along with free-choice starter and water. Calves were weaned on d 42. Jugular blood was sampled at 0800, 1200, and 1600 h, and within 5 min of sampling BHB, and glucose concentrations were estimated using test strips (Nova Max Plus meter, Nova Biomedical Corporation, Waltham, MA). Age effects and time of day were estimated by sampling blood weekly (d 6, 13, 20, 27, 34, 41, and 48). To determine vaccination stress, a Pasteurella vaccine was administered after blood sampling at 0800 h on d 36. Effect of voluntary starter intake was tested by selecting calves for low and high intakes (d 35 to 39) and sampling on d 40, 41, 43, and 44. Starter intake restriction was tested by restricting intake in half of the calves and sampling on d 60 and 61. Data were analyzed with repeated measurements in a mixed model procedure with either within-calf effect (day or week) or within-calf effects (hour, and day or week) included in the model. Time of day did not affect blood BHB and glucose in the first 6 wk. Blood BHB was greater in wk 7 versus wk 1 to 6. Blood glucose was greater in the first 5 wk compared with wk 6 and 7. Blood BHB increased and glucose decreased with increasing starter intake. Blood BHB declined due to vaccination, but glucose was unaffected. Starter intake restriction reduced BHB for 3 d and glucose for 2 d after restriction. Both were affected by time of day. Around weaning (d 40 to 44), BHB and glucose increased with increasing starter intake. In this research, neither blood BHB nor glucose was a good proxy for starter intake. Blood BHB was positively and glucose negatively related to starter intake; however, relationships were weak, variable, and affected by time of day, stress, and intake restriction. Over 30% of calves tested ≤0.2 mmol/L BHB when consuming >1,250 g/d of starter, and test strip increments were 0.1 mmol/L, which represented >25% of the mean blood BHB concentration. In this study, neither blood BHB nor glucose was an acceptable proxy for estimating starter intake.     

A moderate consumption of Côtes du Rhône red wines affects the progression of aortic lesions,and reduces oxidative stress and p22<Superscript>phox</Superscript>NADPH oxidase activation in an experimental model of diet-induced atherosclerosis,according to the vinification process

Côtes du Rhône red wines prepared from both different types of grape and processing were tested in hamsters receiving an atherogenic diet for 12 weeks and a daily gavage with 12% ethanol, red wines from vinification by flash release (GRE FD, SYR FD and BLEND FD), by tanisage (BLEND TAN), by traditional vinification (BLEND), or water as control. Except BLEND TAN, consumption of wines lowered plasma total and LDL-cholesterol. Aortic lipid deposits were reduced by ethanol (30%), wines (54% on average) or BLEND TAN (65%). Cardiac production of superoxide anion decreased from 20% (BLEND and BLEND TAN) to 33% (SYR FD and BLEND FD). The expression of NAD(P)H oxidase decreased by 44% (SYR FD), 26% (BLEND), 47% (BLEND FD) and 44% (BLEND TAN). Except ethanol and GRE FD, wines increased uricemia by 15% on average. These findings indicate that chronic consumption of red wine has potential beneficial effects to prevent the development of atherosclerosis. Prevention of NAD(P)H oxidase induction and preservation of aortic lipids oxidation likely contribute to this effect.