stilbenes and Ischemia

To evaluate the preventive cardioprotective effects of resveratrol and grape products, such as grape juice and red wine, in animal model of cardiac ischemia and reperfusion.. Male Wistar rats orally pretreated for 21-days with resveratrol and grape products were anesthetized and placed on mechanical ventilation to surgically induce cardiac ischemia and reperfusion by obstruction (ischemia) followed by liberation (reperfusion) of blood circulation in left descending coronary artery. These rats were submitted to the electrocardiogram (ECG) analysis to evaluate the effects of pretreatment with resveratrol and grape products on the incidence of ventricular arrhythmias (VA), atrioventricular block (AVB) and lethality (LET) resulting from cardiac ischemia and reperfusion.. It was observed that the incidence of AVB was significantly lower in rats pretreated with resveratrol (25%), grape juice (37.5%) or red wine (12.5%) than in rats treated with saline solution (80%) or ethanol (80%). Similarly, incidence of LET was also significantly lower in rats pretreated with resveratrol (25%), grape juice (25%) or red wine (0%) than in rats treated with saline solution (62.5%) or ethanol (75%).. These results indicate that the cardioprotective response stimulated by resveratrol and grape products prevents the lethal cardiac arrhythmias in animal model of ischemia and reperfusion, supporting the idea that this treatment can be beneficial for prevention of severe cardiac arrhythmias in patients with ischemic heart disease.

Topics: Animals; Arrhythmias, Cardiac; Humans; Ischemia; Male; Rats; Rats, Wistar; Reperfusion; Resveratrol; Stilbenes; Vitis

The impact of maternal nutrition on neurodevelopment and neonatal neuroprotection is a research topic with increasing interest. Maternal diet can also have deleterious effects on fetal brain development. Fetal exposure to alcohol is responsible for poor neonatal global development, and may increase brain vulnerability to hypoxic-ischemic encephalopathy, one of the major causes of acute mortality and chronic neurological disability in newborns. Despite frequent prevention campaigns, about 10% of women in the general population drinks alcohol during pregnancy and breastfeeding. This study was inspired by this alarming fact. Its aim was to evaluate the beneficial effects of maternal supplementation with two polyphenols during pregnancy and breastfeeding, on hypoxic-ischemic neonate rat brain damages, sensorimotor and cognitive impairments, in a context of moderate maternal alcoholism. Both stilbenoid polyphenols, trans-resveratrol (RSV - 0.15 mg/kg/day), and its hydroxylated analog, trans-piceatannol (PIC - 0.15 mg/kg/day), were administered in the drinking water, containing or not alcohol (0.5 g/kg/day). In a 7-day post-natal rat model of hypoxia-ischemia (HI), our data showed that moderate maternal alcoholism does not increase brain lesion volumes measured by MRI but leads to higher motor impairments. RSV supplementation could not reverse the deleterious effects of HI coupled with maternal alcoholism. However, PIC supplementation led to a recovery of all sensorimotor and cognitive functions. This neuroprotection was obtained with a dose of PIC corresponding to the consumption of a single passion fruit per day for a pregnant woman.

Topics: Alcohol Drinking; Alcoholism; Animals; Animals, Newborn; Brain; Brain Injuries; Cognitive Dysfunction; Female; Hypoxia; Hypoxia-Ischemia, Brain; Ischemia; Male; Maternal Nutritional Physiological Phenomena; Maternal-Fetal Exchange; Neuroprotection; Neuroprotective Agents; Polyphenols; Pregnancy; Prenatal Exposure Delayed Effects; Rats; Rats, Wistar; Resveratrol; Stilbenes

Hypoxia-ischemia (HI) remains a major cause of perinatal mortality and chronic disability in newborns worldwide (1-6 for 1000 births) with a high risk of future motor, behavioral and neurological deficits. Keeping newborns under moderate hypothermia is the unique therapeutic approach but is not sufficiently successful as nearly 50% of infants do not respond to it. In a 7-day post-natal rat model of HI, we used pregnant and breastfeeding female nutritional supplementation with piceatannol (PIC), a polyphenol naturally found in berries, grapes and passion fruit, as a neuroprotective strategy. Maternal supplementation led to neuroprotection against neonate brain damage and reversed their sensorimotor deficits as well as cognitive impairments. Neuroprotection of per os maternal supplementation with PIC is a preventive strategy to counteract brain damage in pups induced by HI. This nutritional approach could easily be adopted as a preventive strategy in humans.

Topics: Animals; Animals, Newborn; Behavior, Animal; Brain; Brain Injuries; Cognitive Dysfunction; Dietary Supplements; Disease Models, Animal; Female; Hypoxia; Hypoxia-Ischemia, Brain; Ischemia; Maternal Nutritional Physiological Phenomena; Neurons; Neuroprotection; Neuroprotective Agents; Pregnancy; Rats; Stilbenes

Recent studies highlighted the protective benefits of a Chinese herb extract from polygonum cuspidatum, trans-polydatin, on cardiac disease. We investigated the therapeutic effect of trans-polydatin on myocardial ischemia/reperfusion (IR) injury and the underlying mechanisms related to the renin-angiotensin system (RAS) and RhoA kinase (ROCK) pathway.. Experiments were performed on neonatal rats' ventricular myocytes that were subjected to hypoxia-reoxygenation (simulated IR, SIR) and on adult mice which were subjected to left anterior descending coronary artery occlusion for 45 min followed by a one-week reperfusion. trans-Polydatin significantly increased cell viability and reduced apoptosis in SIR cardiomyocytes. It was also observed to reduce the infarct size and increase the cardiac function in IR mice. trans-Polydatin decreased the expression of angiotensin and inhibited the activities of renin and angiotensin-converting enzyme. Furthermore, trans-polydatin inhibited ROCK activity, especially the angiotensin I receptor-activated ROCK pathway.. trans-Polydatin exerts a cardio-protection against myocardial IR injury likely through inhibiting both RAS and the downstream ROCK pathway.

Topics: Animals; Apoptosis; Glucosides; Heart; Humans; Ischemia; Male; Mice; Mice, Inbred C57BL; Myocardial Reperfusion Injury; Myocardium; Renin-Angiotensin System; rho-Associated Kinases; Signal Transduction; Stilbenes

The natural polyphenol resveratrol (RSV) has been shown to ameliorate ischemia/reperfusion (I/R)-induced damage. Therefore, a rat model of I/R-induced AKI equipped with intensive monitoring was utilized to examine direct renal protection by RSV in vivo.. AKI was induced by bilateral renal clamping (45 min) followed by reperfusion (3 h). Solvent-free RSV was continuously infused intravenously (0.056 and 0.28 mg/kg) in a total volume of 7 ml/kg/h starting from 30 min before renal clamping. At a mean arterial blood pressure below 70 mmHg for more than 5 min, bolus injections of 0.5 ml 0.9% NaCl solution were administered repetitively (max. 5 ml/kg/h).. No differences could be found between normoxic control groups with/without RSV. Bilateral renal clamping and subsequent reperfusion caused a progressive rise in creatinine, cystatin C, and CK, a decrease in cellular ATP content and diuresis. Infusion of RSV increased sirtuin 1 expression after ischemia/reperfusion and was associated with decreased blood pressure during ischemia and early reperfusion accompanied by an increased requirement of bolus injections as well as with increased expression of TNFα.. RSV did not exert protective effects on I/R-induced AKI in the present short-term in vivo rat model. The lack of protection is potentially connected to aggravation of blood pressure instability.

Topics: Acute Kidney Injury; Animals; Blood Pressure; Ischemia; Rats; Reperfusion Injury; Resveratrol; Sirtuin 1; Stilbenes; Treatment Outcome; Tumor Necrosis Factor-alpha

Several studies of stem cell-based gene therapy have indicated that long-lasting regeneration following vessel ischemia may be stimulated through VEGFA gene therapy and/or MSC transplantation for reduction of ischemic injury in limb ischemia and heart failure. The therapeutic potential of MSC transplantation can be further improved by genetically modifying MSCs with genes which enhance angiogenesis following ischemic injury. In the present study, we aimed to develop an approach in MSC-based therapy for repair and mitigation of ischemic injury and regeneration of damaged tissues in ischemic disease. HSP70 promoter-driven VEGFA expression was induced by resveratrol (RSV) in MSCs, and in combination with known RSV biological functions, the protective effects of our approach were investigated by using ex vivo aortic ring coculture system and a 3D scaffolds in vivo model. Results of this investigation demonstrated that HSP promoter-driven VEGFA expression in MSC increased approximately 2-fold over the background VEGFA levels upon HSP70 promoter induction by RSV. Exposure of HUVEC cells to medium containing MSC in which VEGFA had been induced by cis-RSV enhanced tube formation in the treated HUVEC cells. RSV-treated MSC cells differentiated into endothelial-like phenotypes, exhibiting markedly elevated expression of endothelial cell markers. These MSCs also induced aortic ring sprouting, characteristic of neovascular formation from pre-existing vessels, and additionally promoted neovascularization at the MSC transplantation site in a mouse model. These observations support a hypothesis that VEGFA expression induced by cis-RSV acting on the HSP70 promoter in transplanted MSC augments the angiogenic effects of stem cell gene therapy. The use of an inducible system also vastly reduces possible clinical risks associated with constitutive VEGFA expression.

Topics: Animals; Aorta; Bone Marrow Cells; Cell Culture Techniques; Cell Differentiation; Cell Proliferation; Cell Survival; Cells, Cultured; Coculture Techniques; Disease Models, Animal; HSP70 Heat-Shock Proteins; Human Umbilical Vein Endothelial Cells; Humans; In Vitro Techniques; Ischemia; Isomerism; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mice; Mice, Inbred C57BL; Neovascularization, Physiologic; Resveratrol; Stilbenes; Vascular Endothelial Growth Factor A

Resveratrol is an antioxidant agent with multiple positive impacts on the body. It is known to have anti-diabetic, anti-inflammatory, anti-carcinogenic, and neuroprotective effects. The goal of this study is to demonstrate the antioxidant and anti-diabetic effects of resveratrol on flap survival in diabetic rats. Streptozotocin-induced diabetic Sprague-Dawley albino rats were treated with 10 mg/kg resveratrol following a flap surgery. Histological findings regarding polymorphonuclear leukocyte (PMNL) density, vascular proliferation, fibroblast density, and tissue necrosis were compared between resveratrol-treated and control rats. Significantly higher PMNL density was found in the control group (p = 0.005); while vascular proliferation and the fibroblast density were higher in the resveratrol group (p = 0.004 and p = 0.021, respectively). Collagen density was also higher in the resveratrol group and the difference has statistical significance (p = 0.024). Lymphocyte density was not significantly different between groups (p = 0.061). When the necrosis in the distal areas was evaluated histologically, 20% of the resveratrol group had epidermal tissue necrosis, thus 90% of the control group had epidermal or full-layer necrosis. Resveratrol improved flap survival significantly in diabetic rats. Therefore, diabetic patients requiring complex reconstructive procedures may benefit from resveratrol; so, clinical trials are required to support this study.

Topics: Animals; Antioxidants; Diabetes Mellitus, Experimental; Hypoglycemic Agents; Ischemia; Necrosis; Oxidative Stress; Rats, Sprague-Dawley; Resveratrol; Stilbenes; Surgical Flaps; Wound Healing

Ischemia/reperfusion is a leading cause of liver damage after surgical intervention, trauma, and transplantation. It has been reported that the nicotinamide adenine dinucleotide-dependent deacetylase sirtuin 1 attenuates myocardial, cerebral, and renal ischemia/reperfusion damage. This study aimed to investigate the involvement of sirtuin 1-mediated p66shc inhibition in liver ischemia/reperfusion and explore the effect of carnosic acid and ischemic preconditioning on liver ischemia/reperfusion-induced damage.. Laboratory investigation.. University laboratory.. Male Sprague-Dawley rats and HepG2 cells.. The rats were subjected to 45 minutes of ischemia to 70% of the liver, followed by 3-hour reperfusion. The HepG2 cells were subjected to hypoxia/reoxygenation-induced injury.. In the rats with liver ischemia/reperfusion injury, carnosic acid pretreatment and ischemic preconditioning dramatically reduced the serum aminotransferase activity and proinflammatory chemokine levels and improved the liver histological evaluations. Carnosic acid and ischemic preconditioning also increased manganese superoxide dismutase and Bcl-xL, but down-regulated cleaved caspase-3. Interestingly, the protective effect of carnosic acid and ischemic preconditioning was positively associated with sirtuin 1 activation. By contrast, p66shc, a kinase that promotes oxidative injury and apoptosis, was inhibited by carnosic acid and ischemic preconditioning. Sirtuin 1 small interfering RNA knockdown experiments confirmed that carnosic acid increased sirtuin 1-mediated repression of p66shc in HepG2 cells and that the protective effect of carnosic acid against hypoxia/reoxygenation injury was inhibited by the sirtuin 1 inhibitor nicotinamide. These results suggest that carnosic acid protects hepatocytes from hypoxia/reoxygenation damage through sirtuin 1-mediated p66shc suppression. To support this notion, we further demonstrated that the sirtuin 1 activator resveratrol achieved a protective effect similar to that of carnosic acid against hypoxia/reoxygenation injury, whereas sirtuin 1 small interfering RNA and nicotinamide had the opposite effect.. Carnosic acid and ischemic preconditioning protect against ischemia/reperfusion-induced liver injury. Mechanistically, the protective effect involves the sirtuin 1-mediated inhibition of p66shc, suggesting that this pathway is a novel potential therapeutic target for protecting the liver from ischemia/reperfusion injury.

Topics: Abietanes; Analysis of Variance; Animals; Caspases; Enzyme Inhibitors; Hep G2 Cells; Humans; In Situ Nick-End Labeling; Ischemia; Ischemic Preconditioning; Liver; Male; Niacinamide; Plant Extracts; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Resveratrol; RNA, Small Interfering; Shc Signaling Adaptor Proteins; Sirtuin 1; Src Homology 2 Domain-Containing, Transforming Protein 1; Stilbenes; Superoxide Dismutase; Transaminases

Studying human vascular disease in conventional cell cultures and in animal models does not effectively mimic the complex vascular microenvironment and may not accurately predict vascular responses in humans. We utilized a microfluidic device to recapitulate both shear stress and O2 levels in health and disease, establishing a microfluidic vascular model (μVM). Maintaining human endothelial cells (ECs) in healthy-mimicking conditions resulted in conversion to a physiological phenotype namely cell elongation, reduced proliferation, lowered angiogenic gene expression and formation of actin cortical rim and continuous barrier. We next examined the responses of the healthy μVM to a vasotoxic cancer drug, 5-Fluorouracil (5-FU), in comparison with an in vivo mouse model. We found that 5-FU does not induce apoptosis rather vascular hyperpermeability, which can be alleviated by Resveratrol treatment. This effect was confirmed by in vivo findings identifying a vasoprotecting strategy by the adjunct therapy of 5-FU with Resveratrol. The μVM of ischemic disease demonstrated the transition of ECs from a quiescent to an activated state, with higher proliferation rate, upregulation of angiogenic genes, and impaired barrier integrity. The μVM offers opportunities to study and predict human ECs with physiologically relevant phenotypes in healthy, pathological and drug-treated environments.

Topics: Animals; Atherosclerosis; Capillary Permeability; Endothelial Cells; Endothelium, Vascular; Fluorouracil; Humans; Ischemia; Mice; Microfluidic Analytical Techniques; Oxygen Consumption; Resveratrol; Stilbenes; Stress, Mechanical; Vascular Diseases

An in vitro model of ischemic cerebral stroke [oxygen-glucose deprivation (OGD) for 6 h followed by 24 h reoxygenation (R)] with PC12 cells increases Ca(2+) influx by upregulating native L-type Ca(2+) channels and reactive oxygen species (ROS) generation. This reactive oxygen species generation and increase in intracellular Ca(2+) triggers the expression of hypoxic homeostasis transcription factors such as hypoxia induced factor-1 alpha (HIF-1α), Cav-beta 3 (Cav β3), signal transducer and activator of transcription 3 (STAT3), heat shock protein 27 (hsp-27), and cationic channel transient receptor potential melastatin 7 (TRPM7). OGD insulted PC12 cells were subjected to biologically safe doses (5, 10, and 25 μM) of trans-resveratrol in three different treatment groups: 24 h prior to OGD (pre-treatment); 24 h post OGD (post-treatment); and from 24 h before OGD to end of reoxygenation period (whole-treatment). Here, we demonstrated that OGD-R-induced neuronal injury/death is by reactive oxygen species generation, increase in intracellular calcium levels, and decrease in antioxidant defense enzymes. trans-Resveratrol increases the viability of OGD-R insulted PC12 cells, which was assessed by using MTT, NRU, and LDH release assay. In addition, trans-resveratrol significantly decreases reactive oxygen species generation, intracellular Ca(2+) levels, and hypoxia associated transcription factors and also increases the level of antioxidant defense enzymes. Our data shows that the whole-treatment group of trans-resveratrol is most efficient in decreasing hypoxia induced cell death through its antioxidant properties.

Topics: Animals; Antioxidants; Calcium; Calcium Channels; Cell Death; Cell Hypoxia; HSP27 Heat-Shock Proteins; Hypoxia; Hypoxia-Inducible Factor 1, alpha Subunit; Ischemia; L-Lactate Dehydrogenase; PC12 Cells; Rats; Reactive Oxygen Species; Resveratrol; STAT3 Transcription Factor; Stilbenes; Superoxide Dismutase; Superoxide Dismutase-1; TRPM Cation Channels

Accumulating evidence shows that post-ischemic inflammation originated by Toll-like receptors (TLR) plays critical roles in ischemic stroke. However, the functions of other innate immune receptors are poorly understood in cerebral ischemia. Macrophage-inducible C-type lectin, Mincle, is one of the innate immune receptor C-type lectin-like receptor (CLR) to response against dying cells. In the present study, we showed that Mincle, its ligand SAP130, and its downstream phospho-Syk/Syk were upregulated after ischemia, and that Mincle is expressed in immune and non-immune cells in the ischemic brains of mice and human. We treated mice with piceatannol, a Syk inhibitor, and consequently the infarct volume and swelling were suppressed by piceatannol. The levels of phospho-Syk, MMP9 and ICAM-1 were downregulated, and the level of Claudin5 was uplegurated in piceatannol-treated groups. These data indicate that innate immune system, such as Mincle and Syk plays a pivotal role in the pathogenesis after the ischemia and reperfusion.

Topics: Animals; Brain; Down-Regulation; Humans; Immunity, Innate; In Vitro Techniques; Intercellular Adhesion Molecule-1; Intracellular Signaling Peptides and Proteins; Ischemia; Lectins, C-Type; Male; Matrix Metalloproteinase 9; Membrane Proteins; Mice; Protective Agents; Protein-Tyrosine Kinases; Stilbenes; Stroke; Syk Kinase; Up-Regulation

Resveratrol has been demonstrated to be protective in the cardiovascular system. The aim of this study was to assess the effects of resveratrol on hydrogen peroxide (H(2)O(2))-induced increase in late sodium current (I(Na.L)) which augmented the reverse Na(+)-Ca(2+) exchanger current (I(NCX)), and the diastolic intracellular Ca(2+) concentration in ventricular myocytes.. I(Na.L), I(NCX,) L-type Ca(2+) current (I(Ca.L)) and intracellular Ca(2+) properties were determined using whole-cell patch-clamp techniques and dual-excitation fluorescence photomultiplier system (IonOptix), respectively, in rabbit ventricular myocytes.. Resveratrol (10, 20, 40 and 80 µM) decreased I(Na.L) in myocytes both in the absence and presence of H(2)O(2) (300 µM) in a concentration dependent manner. Ranolazine (3-9 µM) and tetrodotoxin (TTX, 4 µM), I(Na.L) inhibitors, decreased I(Na.L) in cardiomyocytes in the presence of 300 µM H(2)O(2). H(2)O(2) (300 µM) increased the reverse I(NCX) and this increase was significantly attenuated by either 20 µM resveratrol or 4 µM ranolazine or 4 µM TTX. In addition, 10 µM resveratrol and 2 µM TTX significantly depressed the increase by 150 µM H(2)O(2) of the diastolic intracellular Ca(2+) fura-2 fluorescence intensity (FFI), fura-fluorescence intensity change (△FFI), maximal velocity of intracellular Ca(2+) transient rise and decay. As expected, 2 µM TTX had no effect on I(Ca.L).. Resveratrol protects the cardiomyocytes by inhibiting the H(2)O(2)-induced augmentation of I(Na.L.)and may contribute to the reduction of ischemia-induced lethal arrhythmias.

Topics: Acetanilides; Animals; Antioxidants; Arrhythmias, Cardiac; Calcium; Diastole; Dose-Response Relationship, Drug; Electrophysiology; Female; Heart Ventricles; Hydrogen Peroxide; Ischemia; Male; Muscle Cells; Patch-Clamp Techniques; Piperazines; Rabbits; Ranolazine; Resveratrol; Sodium; Stilbenes; Temperature; Tetrodotoxin

Implantation of bone marrow-derived mononuclear cells (BMMCs) is known to accelerate blood flow recovery in a hindlimb ischemia model in mice. However, the neovascularization capacity of BMMCs from diabetic mice is impaired. Resveratrol, a natural polyphenolic compound abundant in red wine, is known to extend the lifespan of high cholesterol-fed mice. We tested whether resveratrol improves the neovascularization capacity of BMMCs from diabetic mice. Diabetes was induced by the injection of streptozotocin into C57B/6 mice. BMMCs from normal mice and diabetic mice were implanted into the ischemic limb induced by ligation of the unilateral femoral artery. Blood flow recovery measured by the laser Doppler method was significantly decreased in mice that received BMMCs from diabetic mice compared with BMMCs from normal mice. However, ex vivo treatment of BMMCs from diabetic mice, but not from normal mice, with resveratrol for 30 min significantly improved blood flow recovery. Capillary density measured by PECAM-1 positive cells was significantly increased in mice that received either normal BMMCs or diabetic BMMCs treated with resveratrol. Treatment of BMMCs from diabetic mice with resveratrol increased mRNA expression of vascular endothelial growth factor and endothelial nitric oxide synthase and decreased production of reactive oxygen species. Resveratrol improved the impaired neovascularization capacity of BMMCs derived from diabetic mice. The effects of resveratrol may be due to a reduction of oxidative stress and an induction of angiogenic factors. Resveratrol may be beneficial by improving the neovascularization capacity of BMMCs in patients with diabetes mellitus.

Topics: Animals; Antioxidants; Blood Pressure; Body Weight; Bone Marrow Cells; Capillaries; Diabetes Mellitus, Experimental; Heart Rate; Hindlimb; Ischemia; Laser-Doppler Flowmetry; Malondialdehyde; Mice; Mice, Inbred C57BL; Monocytes; Neovascularization, Physiologic; Nitric Oxide Synthase Type III; Oxidative Stress; Reactive Oxygen Species; Receptors, Vascular Endothelial Growth Factor; Regional Blood Flow; Resveratrol; Reverse Transcriptase Polymerase Chain Reaction; Stilbenes; Superoxide Dismutase; Vascular Endothelial Growth Factor A

To investigate protective effects of polydatin(PD) during lung ischemia/reperfusion in rabbits and its potential mechanisms.. Rabbit lung model of ischemia/reperfusion (I/R) injury was constituted in vivo. Thirty rabbits were divided into groups randomly: Control (C), I/R, PD group, respectively. Endotoxin (ET) in plasma was analyzed by End-point Chromogenic Assay, the expression of Toll-like receptor 4 (TLR4) mRNA, nuclear factor (NF)-kappaBp65 mRNA, intracellular adhesion molecule-1 (ICAM-1) mRNA were measured by RT-PCR, the morphological changes of lung tissue were observed with hematoxylin-eosin (HE) staining.. There was no significant difference in ET concentration of plasma between groups (all of P > 0.05). The expression of TLR-4 mRNA, NF-kappaBp65 mRNA and ICAM-1mRNA in I/R group were significantly increased as compared to C group and PD group, while those expressions in PD group were evidently higher than those in C group (all of P < 0.01). Light microscope showed that the lung pathological injuries in PD group were obviously alleviated as compared to I/R group.. PD might have a protective effect on lung ischemia/reperfusion injury by down-regulating TLR4 and NF-kappaB expression, then inhibiting the release of mediators of inflammation as ICAM-1.

Topics: Animals; Female; Glucosides; Intercellular Adhesion Molecule-1; Ischemia; Lung; Male; Protective Agents; Rabbits; Reperfusion Injury; RNA, Messenger; Signal Transduction; Stilbenes; Toll-Like Receptor 4; Transcription Factor RelA

Moderate consumption of red wine is associated with a decreased incidence of cardiovascular diseases in populations with relatively high amount of fat in the diet. However, the mechanisms involved in this protective effect are not completely understood. Here we show that moderate consumption of red wine (equivalent to 2 glasses/day in humans) but not ethanol only, improves blood flow recovery by 32% after hindlimb ischemia in hypercholesterolemic ApoE-deficient mice. In ischemic tissues, red wine consumption reduces oxidative stress and increases capillary density by 46%. Endothelial progenitor cells (EPCs) have been shown to have an important role in postnatal neovascularization. We found that the number of EPCs is increased by 60% in ApoE mice exposed to red wine. Moreover, the migratory capacity of EPCs is significantly improved in red wine-drinking mice. The wine used in our study is a cabernet sauvignon from Languedoc-Roussillon, France, which contains a relatively high concentration (4-6 mg/L) of the polyphenolic antioxidant resveratrol. We demonstrate that resveratrol can rescue oxidized low-density lipoprotein (oxLDL)-induced impairment of in vitro angiogenic activities in human umbilical vein endothelial cells (HUVECs). Resveratrol exposure is also associated with increased activation of Akt/eNOS together with a restoration of nitric oxide production in HUVECs exposed to oxLDL. Our study suggests that moderate consumption of red wine improves ischemia-induced neovascularization in high-cholesterol conditions by increasing the number and the functional activities of EPCs and by restoring the Akt-eNOS-NO pathway.

Topics: Adult Stem Cells; Animals; Apolipoproteins E; Cell Movement; Cells, Cultured; Endothelium, Vascular; Hindlimb; Humans; Hypercholesterolemia; Ischemia; Mice; Mice, Inbred C57BL; Mice, Knockout; Neovascularization, Pathologic; Nitric Oxide; Resveratrol; Stilbenes; Wine

Thrombopoietic cells may differentially promote or inhibit tissue vascularization by releasing both pro- and antiangiogenic factors. However, the molecular determinants controlling the angiogenic phenotype of thrombopoietic cells remain unknown. Here, we show that expression and release of thrombospondins (TSPs) by megakaryocytes and platelets function as a major antiangiogenic switch. TSPs inhibited thrombopoiesis, diminished bone marrow microvascular reconstruction following myelosuppression, and limited the extent of revascularization in a model of hind limb ischemia. We demonstrate that thrombopoietic recovery following myelosuppression was significantly enhanced in mice deficient in both TSP1 and TSP2 (TSP-DKO mice) in comparison with WT mice. Megakaryocyte and platelet levels in TSP-DKO mice were rapidly restored, thereby accelerating revascularization of myelosuppressed bone marrow and ischemic hind limbs. In addition, thrombopoietic cells derived from TSP-DKO mice were more effective in supporting neoangiogenesis in Matrigel plugs. The proangiogenic activity of TSP-DKO thrombopoietic cells was mediated through activation of MMP-9 and enhanced release of stromal cell-derived factor 1. Thus, TSP-deficient thrombopoietic cells function as proangiogenic agents, accelerating hemangiogenesis within the marrow and revascularization of ischemic hind limbs. As such, interference with the release of cellular stores of TSPs may be clinically effective in augmenting neoangiogenesis.

Topics: Animals; Blood Platelets; Bone Marrow; Chemokine CXCL12; Chemokines, CXC; Hematopoietic Stem Cells; Hindlimb; Ischemia; Megakaryocytes; Mice; Mice, Inbred C57BL; Mice, Knockout; Neovascularization, Pathologic; Neovascularization, Physiologic; Stilbenes; Thrombopoiesis; Thrombopoietin; Thrombospondin 1; Thrombospondins

Recent studies have suggested that an ischemia/reperfusion (I/R) injury enhances the expression of costimulatory adhesion molecules on the vascular endothelium. In the present study, we investigated the protective effects of resveratrol, a phenolic product, on the renal function and expression of CD86 in rat kidneys with I/R injury. Wistar rats were divided into four groups; 1) an I/R group with right nephrectomy and 1-hour clamping of the left renal pedicle; 2) a vehicle group, I/R plus 10% ethanol (0.1 ml/kg/day) administered by intra-peritoneal injection from day -1 through to 7; 3) a resveratrol group, I/R plus 4 mg/kg/day of resveratrol; and 4) a sham group. Blood samples were obtained via the tail vein at 1 day before, and 1, 3, and 7 days after the operation (day 0) for the measurement of serum creatinine (Scr) levels. The expression of CD86 protein was analyzed by immunofluorescence staining, and the level of CD86 messenger RNA (mRNA) was evaluated quantitatively by a real-time reverse transcription-polymerase chain reaction (RT-PCR) in the renal cortex at day 3. Scr levels of the resveratrol group were significantly lower than those of the I/R and vehicle groups on days 1 and 3 after the operation. From the immunohistochemical study, the expression of CD86 in the glomerular endothelium and peritubular vessels was found to be attenuated in the resveratrol group compared with the I/R or vehicle group. In the resveratrol group, the CD86 mRNA level was significantly lower than that in the I/R or vehicle group, and it was significantly decreased by about one fifth of that in the sham group. Our results suggest that resveratrol markedly reduces renal dysfunction and attenuates the mRNA and protein expression of CD86 following I/R injury.

Topics: Animals; Antigens, CD; B7-2 Antigen; Creatinine; Endothelium, Vascular; Ischemia; Kidney; Kidney Cortex; Kidney Function Tests; Kidney Glomerulus; Membrane Glycoproteins; Rats; Rats, Wistar; Reperfusion Injury; Resveratrol; RNA, Messenger; Stilbenes

Ischemia is an inciting factor in 50% of incidences of acute renal failure, and it increases the risk of organ rejection after renal transplantation. We have previously demonstrated that resveratrol (RSV) reduces ischemia-reperfusion (I/R) injury of rat kidney both by antioxidant and anti-inflammatory mechanisms. However, a clear morphological demonstration of this activity has not been made. To answer this question we have performed a new set of experiments following the experimental protocol reported below to investigate the effects of I/R injury and RSV pretreatment on kidney morphology by computerized morphometric analysis. Both renal arteries were clamped for 40 minutes in 40 male Wistar rats (b.w. 220 +/- 20 g); 20 rats were pretreated with RSV 1 microM e.v. 40 minutes before clamping. All animals were reperfused for 24 hours and then sacrificed. Histological examination showed tissue conservation in treated rats. I/R-induced glomerular collapse (as revealed by mean glomerular volume and glomerular shape factor) was significantly reduced by RSV pretreatment. Capillary tuft/Bowman's capsule area ratio was enhanced in the I/R group suggesting tubular hypertension. RSV pre-treatments significantly reduced this parameter to the control value. The number of platelet clots in the capillary tuft and tubular necrosis were also reduced by RSV versus I/R group. L-NAME administration worsened both functional and structural damage. Finally, cGMP urinary levels were markedly reduced from 12.1 +/- 8.4 nmol/day to 0.10 +/- 0.10 nmol/day in the I/R group. RSV provided cGMP (5.01 +/- 1.5 nmol/day, P < 0.05). As expected, L-NAME administration significantly reduced cGMP in urine (0.71 +/- 0.6 nmol/day). The present study confirms the protective effect of RSV pretreatment in I/R injury of rat kidney and suggests multiple mechanisms of action.

Topics: Animals; Cyclic GMP; Dose-Response Relationship, Drug; Ischemia; Kidney Diseases; Kidney Glomerulus; Male; Rats; Rats, Wistar; Renal Circulation; Reperfusion Injury; Resveratrol; Stilbenes; Vitis; Wine

Reactive oxygen species have been implicated in the pathophysiology of renal ischemia reperfusion injury. Antioxidants including polyphenolics have been found to protect renal cells from the cellular injury induced by ischemia and reperfusion. Resveratrol, a stilbene polyphenol found in grapes and red wine, has recently been found to protect isolated rat heart from ischemia reperfusion injury. This study was sought to determine if resveratrol could also protect renal cells from ischemic injury. Male Wistar rats were treated with control, resveratrol (0.23 microg/kg), vehicle used to solubilize resveratrol, and resveratrol plus L-NAME (15 mg/kg body wt), a nitric oxide blocker. Our results demonstrated that resveratrol administration reduced the mortality of ischemic rats from 50% to 10% and renal damage was reduced as indicated by histologic examination and serum creatinine level. The short-term administration of resveratrol also inhibited renal lipid peroxidation induced by ischemia and reperfusion both in cortex and in medulla. Electron paramagnetic resonance detected an increased formation of nitric oxide in the resveratrol-treated kidney that was reduced to the baseline value after treating the rats with L-NAME in addition to resveratrol. The results suggest that resveratrol reduced the renal ischemia reperfusion injury through a nitric oxide-dependent mechanism.

Topics: Animals; Antioxidants; Ischemia; Kidney; Lipid Peroxidation; Male; Nitric Oxide; Rats; Rats, Wistar; Reperfusion Injury; Resveratrol; Stilbenes