C009 Perte du gradient transmural de la fonction mitochondriale et altération du couplage excitation-contraction dans l’insuffisance cardiaque ischémique

L’insuffisance cardiaque (IC) est caractérisée par des altérations du métabolisme énergétique associées à une augmentation de la production de radicaux libres (RL). Les RL altèrent le couplage excitation-contraction (CEC) des myocytes en interagissant avec la signalisation calcique et les protéines contractiles. Chez des rats ayant subit une ligature de l’artère coronaire gauche (PMI), nous avons déterminé si, au stade d’insuffisance cardiaque, la perte du gradient transmural de contractilité et l’altération de la signalisation Ca2+ étaient associées à une dysfonction mitochondriale régionalisée au sein de la paroi du ventricule gauche (VG).Les propriétés métaboliques ont été évaluées en mesurant l’autofluorescence du NADH (microscopie multiphotonique), et les activités de la citrate synthase (CS) et de la cytochrome-c oxydase (COX) de cardiomyocytes isolés du sous-endocarde (ENDO) et du sousépicarde (EPI) du VG de rats PMI ou contrôles (sham). Parallèlement, nous avons mesuré les activités de la superoxyde dismutase (SOD) et de la catalase ainsi que la production mitochondriale de RL (MitoSOX) en microscopie confocale. Le raccourcissement cellulaire, la sensibilité au Ca2+ des myofilaments, le transitoire Ca2+, ainsi que les sparks Ca2+ ont été mesurés en absence ou en présence d’un antioxydant (N-acetyl cysteine NAC: 20mM).Chez les shams, l’utilisation du NADH au cours d’une stimulation électrique est plus importante dans l’ENDO que dans l’EPI et s’accompagne d’activités CS et COX plus élevées. Ce gradient transmural de capacité oxydative disparait au cours de l’IC en raison d’altérations localisées uniquement dans l’ENDO. Ces perturbations métaboliques sont associées à une diminution des défenses antioxydantes et à une élévation de la production de RL dans l’ENDO. Le NAC améliore les propriétés contractiles, la fuite diastolique de Ca2+ du réticulum sarcoplasmique (baisse de la fréquence des sparks spontanés) et réduit le nombre de transitoires Ca2+ ectopiques pro-arythmogéniques dans l’ENDO.En conclusion, la perte du gradient transmural de contractilité au cours de l’IC est partiellement due à une altération régionalisée de la fonction mitochondriale. De plus, la production exacerbée de RL associée aux troubles métaboliques participe à la genèse d’événements arythmiques dans la région sous-endocardique

Plasma Zinc But Not the Exchangeable Zinc Pool Size Differs Between Young and Older Korean Women

This study was done to determine the effect of age on zinc metabolism and status among healthy Korean women. Measures of zinc metabolism and status were measured in eight young women (22–24 years) and seven elderly women (66–75 years) consuming a typical Korean diet. Oral and intravenous tracers highly enriched in 67Zn and 70Zn were administered simultaneously. Multiple plasma, 24-h urines, and fecal samples were collected after isotope administration. In the young women, additional plasma were collected to determine zinc kinetics using a seven-compartmental model. Exchangeable Zinc Pool (EZP) was estimated by Miller’s method. Plasma zinc concentrations were higher in older women than younger women (p < 0.05). EZP and urinary zinc tended to be higher in older women than younger women. Fractional and total zinc absorption and endogenous fecal zinc losses did not differ between young and older women. A comparison of the zinc kinetics of the Korean and American women showed no differences in plasma or EZP zinc parameters. However, absorbed zinc and zinc flux to slowly turning over tissues (Q7) were lower in Korean women than that of Americans (p < 0.01) suggesting the total body zinc content of Korean women is lower than that of American women

Influence of antioxidant (L- ascorbic acid) on tolbutamide induced hypoglycaemia/antihyperglycaemia in normal and diabetic rats

BACKGROUND: Diabetes mellitus is a chronic metabolic disorder characterized by hyperglycaemia. Increased oxidative stress and decreased antioxidant levels are the leading cause of diabetes and diabetic complications. So it is felt that supplementation of antioxidants may be useful in controlling the glucose levels and to postpone the occurrence of diabetic complications. The objective of our study is to find the influence of antioxidant supplementation (L-ascorbic acid) on tolbutamide activity in normal and diabetic rats. METHODS: L- ascorbic acid/tolbutamide/L-ascorbic acid + tolbutamide were administered orally to 3 different groups of albino rats of either sex in normal and diabetic condition. Blood samples were collected from retro-orbital puncture at different time intervals and were analyzed for blood glucose by GOD-POD method. Diabetes was induced by alloxan 100 mg/kg body weight administered by I.P route. RESULTS: L-ascorbic acid/ tolbutamide produced hypoglycaemic activity in a dose dependant manner in normal and diabetic condition. In the presence of L-ascorbic acid, tolbuatmide produced early onset of action and maintained for longer period compared to tolbutamide matching control. CONCLUSION: Supplementation of antioxidants like L-ascorbic acid was found to improve tolbutamide response in normal and diabetic rats

Quantitative Metabolomics by 1H-NMR and LC-MS/MS Confirms Altered Metabolic Pathways in Diabetes

Insulin is as a major postprandial hormone with profound effects on carbohydrate, fat, and protein metabolism. In the absence of exogenous insulin, patients with type 1 diabetes exhibit a variety of metabolic abnormalities including hyperglycemia, glycosurea, accelerated ketogenesis, and muscle wasting due to increased proteolysis. We analyzed plasma from type 1 diabetic (T1D) humans during insulin treatment (I+) and acute insulin deprivation (I-) and non-diabetic participants (ND) by 1H nuclear magnetic resonance spectroscopy and liquid chromatography-tandem mass spectrometry. The aim was to determine if this combination of analytical methods could provide information on metabolic pathways known to be altered by insulin deficiency. Multivariate statistics differentiated proton spectra from I- and I+ based on several derived plasma metabolites that were elevated during insulin deprivation (lactate, acetate, allantoin, ketones). Mass spectrometry revealed significant perturbations in levels of plasma amino acids and amino acid metabolites during insulin deprivation. Further analysis of metabolite levels measured by the two analytical techniques indicates several known metabolic pathways that are perturbed in T1D (I-) (protein synthesis and breakdown, gluconeogenesis, ketogenesis, amino acid oxidation, mitochondrial bioenergetics, and oxidative stress). This work demonstrates the promise of combining multiple analytical methods with advanced statistical methods in quantitative metabolomics research, which we have applied to the clinical situation of acute insulin deprivation in T1D to reflect the numerous metabolic pathways known to be affected by insulin deficiency

The enigma of in vivo oxidative stress assessment: isoprostanes as an emerging target

Oxidative stress is believed to be one of the major factors behind several acute and chronic diseases, and may also be associated with ageing. Excess formation of free radicals in miscellaneous body environment may originate from endogenous response to cell injury, but also from exposure to a number of exogenous toxins. When the antioxidant defence system is overwhelmed, this leads to cell damage. However, the measurement of free radicals or their endproducts is tricky, since these compounds are reactive and short lived, and have diverse characteristics. Specific evidence for the involvement of free radicals in pathological situations has been difficult to obtain, partly owing to shortcomings in earlier described methods for the measurement of oxidative stress. Isoprostanes, which are prostaglandin-like bioactive compounds synthesized in vivo from oxidation of arachidonic acid, independently of cyclooxygenases, are involved in many human diseases, and their measurement therefore offers a way to assess oxidative stress. Elevated levels of F2-isoprostanes have also been seen in the normal human pregnancy, but their physiological role has not yet been defined. Large amounts of bioactive F2-isoprostanes are excreted in the urine in normal basal situations, with a wide interindividual variation. Their exact role in the regulation of normal physiological functions, however, needs to be explored further. Current understanding suggests that measurement of F2-isoprostanes in body fluids provides a reliable analytical tool to study oxidative stress-related diseases and experimental inflammatory conditions, and also in the evaluation of various dietary antioxidants, as well as drugs with radical-scavenging properties. However, assessment of isoprostanes in plasma or urine does not necessarily reflect any specific tissue damage, nor does it provide information on the oxidation of lipids other than arachidonic acid

Honey, a Gift from Nature to Health and Beauty: A Review

Benefits of honey are contributed by the composition of its elements such as glucose, fructose, glucose oxidase, vitamins and phenolic compounds. For health, honey can be used to treat wounds due to the antibacterial activity conferred by the hydrogen peroxide produced by glucose oxidase in honey. Anti-inflammatory, anti-oxidant, deodorizing and tissue regeneration activities in honey also help in the wound healing process. It can also be an alternative sweetener for diabetic patients to ensure compliance to a healthy diet. Moreover, honey exerts several effects such as lowering low density lipids and increasing high density lipids, thus reducing risk of atherosclerosis. In terms of beauty, honey can be used on skin and hair. It moisturizes skin through its natural humectant properties contributed by high contents of fructose and glucose. Honey treats acne on the skin due to its antibacterial activity, anti-inflammatory action and tissue repair. The hair can benefit from honey in such a way that the hair has abundance, and becomes easier to comb. However, there have not been as many studies regarding the use of honey in skin in comparison to its use for health. Therefore, future studies on honey could research its use, action and benefits in both cosmetics and dermatology

The stable isotope use in the exploration of bioavailability and metabolism of magnesium

International audienceMagnesium (Mg) is a biologically essential mineral and Mg deficiency is known to lead to severe biochemical and symptomatic disorders. Radioactive isotopes and, more recently, stable isotopes have been used as research tools to determine intestinal Mg absorption in humans and animals under different nutritional and physiological conditions. Mg isotopes are given orally or orally plus intravenously and analysed in faeces and/or in plasma and urine in order to calculate intestinal Mg absorption and possibly endogenous Mg excretion. Mg isotopes have been also used to assess exchangeable pools of Mg under nutritional and physiopathological conditions. Mg isotopes are given intravenously and analysed in plasma and urine to calculate the size and half-life of the various Mg exchangeable pools. Finally, Mg isotopes have been used to study the mechanisms of Mg cellular exchange. To this end, erythrocytes or other types of cells are loaded with Mg isotope or incubated in an isotope-rich medium in order to study the Mg flux and its mechanisms. This paper is a report on the use of stable Mg isotopes and their advantages in these different fields of Mg absorption and metabolism. The studies available have clearly demonstrated that stable isotopes provide a useful research tool for determining intestinal Mg absorption, and represent a precious research tool for the study of Mg metabolism and the assessment of Mg status

Dietary inulin intake and age can significantly affect absorption of the faecal marker dysprosium in rats

International audienceIt is believed that rare earth elements are not absorbed, and thus they are generally used in some mineral absorption studies as a faecal marker. The aim of the present study was to determine the effect of inulin intake and age on dysprosium (Dy) absorption in rats. Eighty male Wistar rats of four different ages (2, 5, 10 and 20 months) were randomised into either a control group or a group receiving 3.75 % inulin in their diet for 4 d and then 7.5 % inulin until the end of the study. The animals were fed fresh food and water ad libitum for 30 d. The intestinal absorption of Dy was determined from a 4 d (day 21 to day 25) balance study. Mean faecal Dy recovery (%) in the eight groups (3 months control, 3 months inulin, 6 months control, 6 months inulin, 11 months control, 11 months inulin, 21 months control, 21 months inulin) was 94.0 (sd 8.6), 64.8 (sd 10.1), 95.8 (sd 9.4), 81.5 (sd 12.1), 98.4 (sd 9.8), 87.8 (sd 9.5), 97.8 (sd 6.2) and 84.9 (sd 10.9), respectively. Our results showed clearly that dietary inulin intake decreased faecal Dy recovery in all four rat groups, and faecal Dy recovery was significantly higher in the old rats (10 and 20 months) than in the young and adult rats. These results show that the faecal recovery (or intestinal absorption) of Dy may vary greatly with nutritional or physiological states such as inulin intake or age. The use of rare earth elements as a faecal marker should be thus validated under each nutritional or physiological state before being employed in mineral absorption studies

Stable isotopes in studies of intestinal absorption, exchangeable pools and mineral status: The example of magnesium

International audienceMagnesium (Mg) is a biologically essential mineral and Mg deficiency is known to lead to severe biochemical and symptomatic disorders. Radioactive isotopes and, more recently, stable isotopes have been used as research tools to determine intestinal Mg absorption in humans and animals under different nutritional and physiological conditions. Mg isotopes are given orally or orally plus intravenously and analysed in faeces and/or in plasma and urine in order to calculate intestinal Mg absorption and possibly endogenous Mg excretion. Mg isotopes have been used to assess exchangeable pools of Mg under nutritional and physiopathological conditions. Mg isotopes are given intravenously and are analysed in plasma and urine to calculate the size and half-life of the various Mg exchangeable pools. More recently, in vitro isotopic tests have been developed to study the need of cells for Mg in different nutritional and genetic conditions. Whole blood is incubated with Mg isotopes and isotopic blood cell enrichment is measured, which reflects the avidity of cells for Mg and thus its initial status. This paper is a report on the use of stable Mg isotopes and their advantages in these different fields of Mg absorption and metabolism. The studies available have clearly demonstrated that stable isotopes provide a useful research tool for determining intestinal Mg absorption, and represent a precious research tool for the study of Mg metabolism and the assessment of Mg status

Dietary inulin intake and age can affect intestinal absorption of zinc and copper in rats

International audienceNondigestible inulin-type fructan intake can stimulate intestinal mineral absorption in both humans and animals. However, this stimulatory effect may depend on experimental conditions such as the duration of the experience, mineral levels in the diet, and the animal's physiological status. The aim of this study was to determine the effect of inulin intake on Zn and Cu absorption in rats at different ages. Male Wistar rats (n = 80) of 4 different ages (2,5, 10, and 20 mo) were randomly assigned to a control group or a group administered 3.75% inulin in their diet for 4 d followed by 7.5% inulin for 26 d. Absorption of Zn-67 and CU65 was determined on d 21 of the experiment by fecal monitoring using Zn-67 and CU65 isotopes. Zn and Cu status was also assessed. Absorption of Zn-67 and CU65 was significantly lower in 11- and 21-mo-old rats than in 3- and 6 mo-old-rats. Moreover, inulin intake significantly increased Zn-67 and CU65 absorption. In conclusion, age and dietary inulin intake can significantly affect intestinal absorption of zinc and copper in rats. Further studies are required to explore this effect over longer periods of inulin intake and to test the effects of inulin in humans