Local Ischemia-reperfusion Research Articles

Involvement of apoptosis related proteins in the neuro-protective effect of dihydromyricetin in transient local ischemia reperfusion injury in rats

Neuronal apoptosis plays a crucial role in mediating cell death after ischemia reperfusion injury. Dihydromyricetin (DHM), a flavonoid, has shown to be neuro-protective by its anti-oxidant, anti-inflammatory and anti-apoptotic properties. The present study investigates whether DHM treatment exerts neuro-protection by modulating apoptosis related proteins after ischemia reperfusion injury. Male Wistar rats (270 ± 20 g) were used after ethical approval. Middle cerebral artery was occluded for 90 min using a silicon coated doccol sutures followed by reperfusion for 3 days. DHM (100 mg/kg) was administered immediately and 2 h after reperfusion followed by single dose every 24 h for next 2 days. On day 3, rats were sacrificed, brain samples homogenised in RIPA buffer and expression of apoptosis related proteins like Bcl2 and Bax were checked by western blot. For apoptosis, rats were euthanized, perfusion fixed with paraformaldehyde, cryosections were stained with TUNEL assay kit. The results indicated that compared to sham group, there were significantly (p<0.001) higher number of TUNEL positive cells indicating more apoptosis in middle cerebral occlusion (MCAo) group. Treatment with DHM significantly (p<0.05) reduced the number of apoptotic cells when compared to MCAo group. The expression of Bax was also increased in MCAo group as compared to sham. DHM treatment significantly normalised the expression of Bax when compared with MCAo. MCAo induced decreased Bcl2 expression was also normalised by DHM. Our results thus showed that the neuro-protective effect of DHM in ischemia reperfusion injury may be due to normalization of apoptosis related proteins.

Compartment syndrome: pathophysiology, clinical presentations, treatment, and prevention in human and veterinary medicine

To review the human and veterinary literature pertaining to all forms of compartment syndrome (CS). Data sources included scientific reviews and original research publications from the human and veterinary literature. While CS affecting the extremities has been recognized in people for decades, other forms of CS in the abdominal and thoracic cavities are recently gaining more attention. The role of CS in critically ill people is a rapidly growing area of interest. More research on prevention and treatment of CS is being conducted in people because some studies have found mortality rates as high as 80% for those suffering from these conditions. While a significant amount of experimental studies of CS have been performed on small animals, there is a marked lack of primary veterinary studies. The majority of the veterinary literature includes case reports and series, and many of these studies were published over a decade ago. However, the increased recognition of CS in people has sparked an interest in veterinary critical care medicine and this has been demonstrated by the recent increased evaluation of compartment pressures in veterinary patients. CS is a complex clinical condition where increased pressure within a compartment can cause significant adverse effects within the compartment as well as throughout the body. Systemic inflammatory responses and local ischemia-reperfusion elements can contribute to the detrimental effects seen in CS. This cascade of events results in increased mortality rates and contributes to the development of CS elsewhere. A better understanding of CS will help veterinarians improve patient care and outcome. Future studies on incidence, prevention, and treatment of CSs in the critical care patient are needed in veterinary medicine.

Remote ischemic preconditioning will reduce graft-related events after surgical infrainguinal revascularization: Evidence-based surgery hypothesis

Surgical lower extremity revascularization obligates ischemia-reperfusion, and local ischemia-reperfusion provokes systemic inflammation; perhaps counterintuitively, remote ischemic preconditioning elicits a constructive anti-inflammatory response. So, remote ischemic preconditioning will decrease graft-related complications after surgical infrainguinal revascularization.

Pulmonary Perfusion With L-Arginine Ameliorates Post-Cardiopulmonary Bypass Lung Injury in a Rabbit Model

Post-cardiopulmonary bypass (CPB) lung injury is the combination of whole body inflammatory response and local ischemia-reperfusion (IR) injury. We investigated the benefit of pulmonary perfusion with L-arginine in protection against post-CPB lung injury. New Zealand white rabbits (n = 50, weight, 2.5-2.8 kg) were divided into five groups (n = 10 each): sham (sham sternotomy), CPB (CPB without pulmonary perfusion), perfusion (CPB with pulmonary perfusion), L-arginine (CPB with perfusion + L-arginine), and L-NAME (CPB with perfusion + L-NAME). The duration of CPB was 60 min followed by 2 h of reperfusion. Pulmonary perfusion was performed every 20 min through the pulmonary artery during CPB. Checking parameters included: (1) pulmonary vascular resistance, (2) pulmonary artery endothelium relaxation (organ chamber study), and (3) IR marker (myeloperoxidase) and inflammatory markers (TNF-α, IL-B, NF-κB). CPB induced pulmonary artery endothelium dysfunction manifested by increased pulmonary vascular resistance and impaired pulmonary artery relaxation. Pulmonary perfusion could significantly reverse the phenomenon (P < 0.01) while provision of NO precursor-L-arginine with pulmonary perfusion together further possessed significant relaxation ability for pulmonary arterial endothelium compared with perfusion alone (P < 0.05). Accordingly, lung parenchyma myeloperoxidase activity and inflammatory cytokine level were also markedly increased after CPB (P < 0.05). Pulmonary perfusion could partially decrease the response, whereas additional L-arginine further attenuated inflammatory cytokine release (P < 0.05). Pulmonary perfusion during CPB partially ameliorates CPB-induced lung injury. Pulmonary perfusion with L-arginine could further attenuate lung injury by restoring endothelial function and decreasing inflammatory response.

Role of Lipoxygenases and Lipoxin A4/Annexin-1 Receptor in Gastric Protection Induced by 20% Ethanol or Sodium Salicylate in Rats

The role of cyclooxygenases and prostaglandins in experimental models of gastroprotection is well established. We investigated the effects of the 5-lipoxygenase inhibitor A63162, the 12-lipoxygenase inhibitor baicalein and the 15-lipoxygenase inhibitor PD146176 as well as the nonspecific lipoxin A₄/annexin-1 antagonist Boc1 on adaptive protection induced by 20% ethanol against 70% ethanol, and on protection induced by sodium salicylate against the mucosal-damage-aggravating effects of celecoxib and dexamethasone during local ischemia-reperfusion in rats. It was found that both types of gastroprotection were antagonized by the lipoxygenase inhibitors and the lipoxin A₄/annexin-1 antagonist in doses that have no direct damaging effect on gastric mucosa. The results suggest that not only cyclooxygenases, but also active lipoxygenases and, possibly, annexin-1 are required for these types of gastroprotection to occur.

Sequential Limb Ischemia Demonstrates Remote Postconditioning Protection of Murine Skeletal Muscle

Ischemic postconditioning, the process of exposing tissues to brief cycles of ischemia-reperfusion after critical ischemia, can mitigate local ischemia-reperfusion injury. Remote protection of skeletal muscle has never been demonstrated in postconditioning models of ischemia-reperfusion injury. Mice were subjected to 2 hours of ipsilateral hind limb ischemia followed by reperfusion. Contralateral limb ischemia was subsequently induced for 2 hours after either 0 (n = 6), 20 (n = 6), or 120 (n = 5) minutes of ipsilateral limb reperfusion. These groups were compared with animals subjected to bilateral simultaneous injury (n = 8) and sham animals that did not undergo ischemia (n = 6). The gastrocnemius muscles were harvested for histologic evaluation, and injury was recorded as the percentage of injured fibers. The first limbs undergoing injury in the 20-minute interval group had a 59 percent injury reduction compared with contralateral limbs (16.0 +/- 2.4 percent versus 39.5 +/- 6.5 percent) after 24 hours of reperfusion and 62 percent reduction after 48 hours (24.4 +/- 3.0 percent versus 63.6 +/- 5.5 percent). In animals with no interval or a 120-minute interval between the onset of limb ischemia, there was no significant difference in injury between hind limbs. The injury in these groups was similar to that in hind limbs subjected to simultaneous bilateral ischemia. A 20-minute reperfusion interval between hind limb ischemia significantly protects against injury in the initially ischemic limb, while similar injury is observed with simultaneous ischemia or an interval of 120 minutes. This study demonstrates remote postconditioning of skeletal muscle and may lead to the development of post hoc therapies.

The influence of iloprost on acute lung injury induced by hind limb ischemia-reperfusion in rats

The local ischemia-reperfusion (I/R) process gains a systemic nature and affects distal organs. The remote effects of I/R are most frequently observed in the lungs and pulmonary damage may vary from acute lung injury with mild dysfunction to severe respiratory failure or the acute respiratory distress syndrome. In this hind limb I/R induced experimental lung injury model two groups of rats as IR and ILO were determined. Both groups underwent 60 min of ischemia and 120 min of reperfusion. While ILO group received iloprost in saline, IR group received only saline before reperfusion period intravenously. Serum myeloperoxidase (MPO) activity, malondialdehyde (MDA) levels and total antioxidant capacity (TAC) and lung tissue MPO activity, MDA levels and Na +–K + ATPase activity were measured and light microscopic analyses of lung specimens were performed. The MPO activities in serum and lung homogenates were found to be significantly decreased in ILO group ( P≤0.01). The MDA levels in lung homogenates were found to be significantly decreased in ILO group ( P≤0.01), but the decreases were not significant in serum MDA levels ( P=0.052). Serum TAC and lung tissue Na +–K + ATPase activity levels were found to be increased in ILO group compared to IR group ( P≤0.01). Lung histology showed marked improvement by iloprost compared to the IR group in this study. Iloprost has been found to be effective in attenuating ischemia reperfusion-induced remote organ damage, in this case, lung injury, in rats.

Role of Endogenous Opioid Receptor Agonists in Regulation of Heart Resistence to the Arrhythmogenic Action of Short-Term Ischemia and Reperfusion

Preliminary selective block of mu-, delta1-, delta2-, and kappa-opioid receptors had no effect on the incidence of ventricular arrhythmias during 10-min coronary occlusion-reperfusion in ketamine-narcotized rats. Repetitive short-term immobilization of rats for 2 weeks improved heart resistance to the arrhythmogenic action of coronary occlusion and reperfusion. Selective mu-opioid receptor antagonist CTAP completely abolished, while selective delta- and kappa-opioid receptor antagonists did not modulate the antiarrhythmic effect of adaptation. Probably, endogenous agonists of mu-opioid receptors play an important role in the adaptive improvement of heart resistance to arrhythmogenic factors, but are insignificant for the modulation of heart resistance to the arrhythmogenic action of short-term local ischemia-reperfusion in non-adapted animals.

Activation of poly(ADP-ribose) polymerase in circulating leukocytes during myocardial infarction.

Myocardial ischemia-reperfusion can lead to increased oxidative stress both locally and in circulating leukocytes. Oxidant-mediated DNA single strand breaks are known to activate the nuclear enzyme poly(ADP-ribose) polymerase (PARP) in various forms of shock, inflammation, and ischemia-reperfusion injury. The aim of the current study was to investigate whether a local insult such as myocardial ischemia-reperfusion is sufficient to lead to activation of PARP in circulating leukocytes. In anesthetized rats myocardial ischemia-reperfusion was induced by transient ligation of the left anterior descending coronary artery. There was a marked increase in poly(ADP-ribosyl)ation of proteins in homogenates of leukocytes isolated from rats at the end of the reperfusion period. Poly(ADP-ribosyl)ation was inhibited by administration of the pharmacologic PARP inhibitor INO-1001 (30 mg/kg) to the rats. We conclude that local insults, such as myocardial reperfusion injury, are sufficient to activate PARP in circulating leukocytes. PARP activation in circulating cells may mediate certain systemic effects of local ischemia-reperfusion injury such as inflammatory mediator production and remote organ injury.

Hepatic Ischemia Is Associated with an Increase in Liver Parenchyma Nitric Oxide That Is in Part Enzyme-Independent

Nitric oxide (NO) might be involved in liver response to local ischemia-reperfusion injury. A specific NO-sensitive electrode was inserted into liver parenchyma of anesthetized rabbits. After a 45-min period of stable NO signal, the vascular pedicle of the caudal lobe of the liver was clamped for 45 min, then the clamp was removed. Perfusion of the right upper lobe was left unchanged. The same procedure was applied in other animals after administration of a long-acting nonspecific NO synthase inhibitor NAPNA. Occlusion of the caudal pedicle was associated with a mean threefold increase in NO signal measured in the caudal lobe. After unclamping, this signal returned within 8 min to baseline value and remained stable for the next 6 h. In the right upper lobe, NO signal was unaffected by caudal lobe ischemia. By the end of the 6-h reperfusion period, administration of the NO inhibitor l-NAME led to a suppression of the NO signal, thus demonstrating the specificity of the measurement. Plasma nitrate and nitrite concentrations remained almost unchanged during the study period in all groups. In animals whose NO synthases had been previously inhibited by NAPNA, clamping the caudal pedicle for 45 min was still associated with a significant increase in caudal lobe NO signal. Nitric oxide is present in liver parenchyma, and its generation is dramatically affected by an ischemia injury. The increased NO generation during local ischemia is, at least in part, independent of NO synthases.

Mast cell mediation of muscle and pulmonary injury following hindlimb ischemia-reperfusion.

We have observed extensive mast cell degranulation in the reperfused hindlimb muscle of the mouse, accompanied by pathological changes within the muscle. As quantitated by the tissue:blood (125)I permeability ratio, both the hindlimbs and lungs exhibited a significant increment in permeability during hindlimb reperfusion. In lungs of the same mice, mast cell-derived chymase mMCP-1 coats alveolar macrophages, an event noted by us in acid-induced direct lung injury. Mast cells in the lung contain mMCP-1, whereas those in the muscle do not. Neither extensive muscle injury nor an increased pulmonary permeability index occurs in the mast cell-deficient W/W(v) mice. We conclude that the mast cell is a key mediator in both local ischemia-reperfusion injury (I-R) of muscle and consequent remote lung injury.

Gastric mucosal injury induced by local ischemia-reperfusion in rats. Role of endogenous endothelin-1 and free radical.

We investigated the role of an endogenous vasoconstrictor peptide endothelin-1 (ET-1) and free radicals in local gastric ischemia-reperfusion injury in rats. Local gastric ischemia was induced by clamping the left gastric artery for 15 min and reperfusion was done for 10-30 min in the presence of 150 mM exogenous HCl intragastrically. Local gastric ischemia and reperfusion resulted in significant macroscopic and microscopic gastric mucosal damage together with elevation of gastric tissue ET-1 concentration. Gastric tissue ET-1 was found to increase after 15 min of ischemia alone and also with 30 min of reperfusion. A novel nonpeptide endothelin receptor antagonist, bosentan, or a combination of radical scavengers (superoxide dismutase, catalase, and deferoxamine) both attenuated gastric mucosal injury. However, the greater protection observed with bosentan than with radical scavengers might reflect a preferential role of endothelin-1 in this type of injury.

Gastric epithelial damage induced by local ischemia-reperfusion with or without exogenous acid

We evaluated the damage to the gastric epithelium produced by local ischemia-reperfusion (IR) with or without luminal perfusion with 0.1 N HCl. Local gastric ischemia was induced by clamping the left gastric artery. Use of radioactive microsphere technique revealed a significant reduction in blood flow induced only in the corpus (67% reduction). Because no measurable gross lesion was observed in this model, the blood-to-lumen clearance of 51Cr-labeled EDTA (51Cr-EDTA) served as an index of epithelial damage. In the absence of exogenous acid, the histological damage was minimum and could not be quantified. However, a significant increase in 51Cr-EDTA clearance was observed shortly after reperfusion in a manner that depended on the duration of ischemia. This increase in clearance reached a maximum approximately 10 min after reperfusion and returned rapidly toward control levels within 40-50 min after reperfusion. In the presence of exogenous acid, EDTA clearance increased significantly during ischemia, increased further during reperfusion, and did not recover for at least 60 min after reperfusion. The acid infused after reperfusion (no acid before reperfusion) did not significantly aggravate the mucosal damage that followed reperfusion. However, the acid infused before reperfusion (no acid after reperfusion) showed an effect on EDTA clearance similar to that induced by continuous acid perfusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Leukocyte adhesion in local versus hemorrhage-induced ischemia.

The objective of this study was to compare the leukocyte-endothelial cell adhesive interactions elicited in postcapillary venules by either local ischemia-reperfusion or hemorrhage-reperfusion. Leukocyte rolling, adherence, and emigration were monitored in cat mesenteric venules exposed to an 85% reduction in blood flow (induced by either hemorrhage or local restriction of arterial inflow) for 1 h, followed by 1 h reperfusion. Leukocyte-endothelial cell interactions, venular diameter, and red blood cell velocity were measured during baseline, ischemia, and reperfusion periods. Both local and hemorrhage-induced ischemia reperfusion caused a reduction in leukocyte rolling velocity and increases in leukocyte adherence and emigration. Quantitatively, the adherence and emigration responses in both ischemia models were nearly identical. However, the two models differed in their response to immunoneutralization of the leukocyte adhesion glycoprotein CD11/CD18 with monoclonal antibody (MAb) IB4. The MAb had a more profound effect in attenuating leukocyte adherence and emigration in the local ischemia model. These results indicate that different factors may contribute to leukocyte-endothelial cell adhesive interactions observed in local vs. systemic models of ischemia-reperfusion.