Effect of sevoflurane preconditioning on ischaemia/reperfusion injury in the rat kidney in vivo

BACKGROUND AND OBJECTIVE: Whereas the protective effect of anaesthetic and ischaemic preconditioning has been described for several organs, it is uncertain whether this mechanism is also effective in the kidney. We compared the effect of preconditioning with sevoflurane and preconditioning with short episodes of ischaemia on renal ischaemia/reperfusion injury in the rat in vivo. METHODS: Fourteen days after right-sided nephrectomy, anaesthetized male Wistar rats were randomly assigned to a sham-operated group (no arterial occlusion, n = 5) or underwent 45 min of left renal artery occlusion (control group, n = 9) followed by 3 days of reperfusion. Two further experimental groups of animals were preconditioned prior to ischaemia either by administering 1 MAC sevoflurane for 15 min followed by 10 min of washout (sevoflurane group, n = 10) or by subjecting the animals to three short episodes of renal ischaemia (ischaemia-preconditioned group, n = 8). Blood creatinine was measured during reperfusion and morphological damage was assessed by histological examination. RESULTS: Baseline creatinine values were similar in all four groups (0.7 +/- 0.2 mg dL-1; mean +/- SD) and remained unchanged in the sham-operated animals after 3 days (0.8 +/- 0.2 mg dL-1). Creatinine levels increased in the ischaemic preconditioning group (3.3 +/- 1.2 mg dL-1) and sevoflurane preconditioning group (4.0 +/- 1.1 mg dL-1) compared to the control group (1.6 +/- 0.6 mg dL-1). Morphological damage was less severe in the control group, i.e. in animals without preconditioning, than in both preconditioning groups. CONCLUSION: Neither sevoflurane nor ischaemic preconditioning preserves renal function or attenuates cell damage in the rat in vivo.     

Effect of free radical scavenging on skeletal muscle blood flow during postischaemic reperfusion.

After 6-h tourniquet ischaemia of one hindlimb in male Sprague-Dawley rats, gastrocnemius muscle blood flow was measured following 10, 120 and 240 min of reperfusion using radiolabelled microspheres. A perfusion index was calculated (experimental limb: contralateral limb) for each of these times. Comparison of perfusion indices in ten control animals (6 h ischaemia, 4 h reperfusion) with similar measurements in ten normal rats with no ischaemia and in ten ischaemic animals with the tourniquet in situ demonstrated low median (interquartile range (i.q.r.)) reflow after 10 min (control 0.12 (0.02-0.43), ischaemia 0.04 (0.00-0.07), normal 1.05 (0.68-1.18); control versus ischaemia, P not significant; control versus normal, P < 0.01). Relative reperfusion occurred at 120 min (control 0.48 (0.11-0.70), ischaemia 0.02 (0.01-0.07), normal 0.97 (0.79-1.13); control versus ischaemia, P < 0.05; control versus normal, P < 0.05) and reperfusion injury after 240 min of revascularization, with muscle blood flow being little different from that in the ischaemic group (control 0.05 (0.01-0.38), ischaemia 0.03 (0.00-0.07), normal 1.01 (0.73-1.16); control versus ischaemia, P not significant; control versus normal, P < 0.01). Two groups of 12 rats were given either intravenous superoxide dismutase and catalase or dimethylthiourea 30 min before tourniquet release, continuing throughout the period of reperfusion. Superoxide dismutase and catalase reversed low reflow, producing a median (i.q.r.) perfusion index of 0.94 (0.54-1.12) (P < 0.01 versus control, P not significant versus normal), but had no effect on relative reperfusion (0.66 (0.42-1.01), P not significant versus control) or on reperfusion injury (0.27 (0.01-0.35), P not significant versus control). In contrast, dimethylthiourea had no effect on perfusion at either 10 min (0.10 (0.03-0.15), P not significant versus control) or 240 min (0.04 (0.00-0.11), P not significant versus control), but abolished the phase of relative reperfusion at 120 min (0.04 (0.02-0.21), P < 0.01 versus control). These results indicate that, although superoxide radicals are harmful during postischaemic reperfusion, hydroxyl radicals may be beneficial.     

Oxygen free radical regulation of rat splanchnic blood flow.

BACKGROUND. The role of oxygen-derived free radicals on splanchnic prostaglandin (PGI2) synthesis and superior mesenteric artery (SMA) blood flow was examined during acute hemorrhage. METHODS. Sprague-Dawley rats were hemorrhaged to 30 mm Hg for 5, 30, or 45 minutes without (SK5, SK30, and SK45) or with (SK5 + R, SK30 + R, and SK45 + R) blood reperfusion. The SK30 + R and SK45 + R groups were treated with or without superoxide dismutase (10,000 units/kg intravenously). In vivo SMA blood flow was measured continuously for 100 minutes by a transonic flow probe, or in other groups the in vitro-perfused SMA and its end-organ intestine (SV+SI) were assayed for release of PGI2 by radioimmunoassay after 15 and 30 minutes of perfusion. RESULTS. Acute hemorrhage at all time periods increased SV+SI PGI2 release twofold to threefold compared with sham animals (p less than 0.01), which was abolished after blood reperfusion. SMA blood flow was decreased by 79.6% +/- 3.3% and 88.2% +/- 1.4% in the SK30 + R and SK45 + R groups compared with the sham animals (p less than 0.01). Superoxide dismutase treatment restored both SV+SI release of PGI2 after SK and SMA blood flow to control levels. CONCLUSION. Oxygen-derived free radicals locally produced during SK30 + R and SK45 + R inhibited splanchnic PGI2 synthesis, which contributed to decreased splanchnic blood flow.     

Epithelial cells and adipose cells both have their own temporal profile in 72-kd heat-shock protein expression determining their tolerance for ischaemia.

Seventy-two-kd heat-shock protein (HSP72) is one of the stress markers induced in cells under stress, such as in the case of ischaemia. Recent studies have suggested that HSP72 is a 'molecular chaperone' to protect cells from various kinds of stress, and that the temporal profile of HSP72 induction is related to ischaemic vulnerability. In this study, we attempted to analyse the temporal profiles of HSP72 induction in epithelial and adipose cells in skin flaps after various periods of transient ischaemia, and we investigated the reason why there were differences in ischaemic tolerance between these cells. We used the abdominal skin flap of Wister rats, which were divided into three groups: the sham control group (n=27), the 2-h ischaemia group (n=25), and the 8-h ischaemia group (n=25). At periods of 8, 24, 48, 96 h, and 7 days after reperfusion, we examined them for any histological changes and performed immunostaining for HSP72 (n=5, each time point). Two animals in the sham control group were sacrificed to harvest the samples immediately after the skin flaps were elevated. As a result, the epithelial cells in all groups revealed positive for HSP72 through the time course, regardless of the ischaemic stresses, and they were alive at 7 days. In the adipose cells, the cells in the sham control group revealed no immunoreactivity after the reperfusion, and they had no change at 7 days. In the 2-h ischaemia group, the adipose cells gradually increased the reactivity for HSP72; consequently they survived beyond 7 days. In the 8-h ischaemia group, the reactivity for HSP72 gradually decreased; consequently they played out a delayed cell death at 7 days. We concluded that these differences of HSP72 expression were related to the cellular vulnerability to ischaemia.     

Prevention of cold ischaemia-reperfusion injury by an endothelin receptor antagonist in experimental renal transplantation.

BACKGROUND: Endothelin (ET) is known to play a role in the pathogenesis of warm ischaemic renal damage, however, little is known about its involvement in renal cold ischaemia. This study was designed to investigate the response of ET after kidney cold ischaemia, and to assess the potential protective effect of bosentan, a dual, non-selective ET(A)/ET(B) receptor antagonist, against cold ischaemia reperfusion injury in a rat model of syngeneic renal transplantation. METHODS: Kidneys from Lewis rats were transplanted, either immediately or after 5 h of cold preservation. After 48 h, contralateral nephrectomy was performed. Rats were organized into three groups: Tr-NoISC, no cold ischaemia; Tr-ISC, 5 h cold ischaemia; and Tr-BOS, 5 h cold ischaemia plus bosentan (100 mg/kg/day, from the day before transplantation until the seventh day post-transplantation). On day 7, plasma and tissue immunoreactive ET (irET), as well as ET mRNA tissue expression, were evaluated. Renal function was measured by means of serum creatinine on days 3, 4, 5 and 7, and by creatinine clearance on day 7. Conventional histology was performed. RESULTS: The ischaemic group had significantly higher plasma irET levels than the non-ischaemic group and significantly lower levels than the bosentan group. Tissue irET levels and ET mRNA expression were similar in the ischaemic and bosentan groups and were higher than in the non-ischaemic group. Throughout the follow-up, serum creatinine was significantly higher in the ischaemic group than in the bosentan group. Moreover, creatinine decreased rapidly in the bosentan group after nephrectomy, whereas it continued to increase for 48 h in the ischaemic group. Kidneys from the ischaemic group showed a higher degree of tubular-cell necrosis and epithelial-cell detachment than kidneys from the bosentan group. CONCLUSIONS: We conclude that cold ischaemia and preservation damage induces an increase in renal ET mRNA and irET expression in the reperfusion phase, contributing both to the deterioration of renal function and to tubular necrosis. Bosentan is effective in protecting kidneys from this cold ischaemia reperfusion damage. Non-selective ET(A)/ET(B) receptor antagonists might be potentially useful in clinical renal transplantation.     

The triphasic pattern of skeletal muscle blood flow in reperfusion injury: an experimental model with implications for surgery on the acutely ischaemic lower limb

A new model of acute lower limb ischaemia has been developed in the rat hind limb. The model has been used to examine the influence on skeletal muscle blood flow of increasing periods of ischaemia and reperfusion. Following restoration of blood flow after 2-6h ischaemia initial gastrocnemius muscle blood flow was reduced. The severity of initial low reflow, indicated by mean perfusion ratios at 0 and 10 min, correlated with the duration of preceding ischaemia (r = 0.83, p less than 0.05) and was greatest in limbs subjected to 6h ischaemia (p less than 0.01 vs. controls at 0, 10 and 60 min after revascularisation). After 60-240 min, reperfusion muscle blood flow returned to normal in limbs subjected to 3, 4 or 5 h ischaemia. In contrast, peak muscle blood flow in limbs that had been ischaemic for 6 h occurred after 120 min, although perfusion remained less than that measured in control limbs (p less than 0.05:6 h ischaemia, 120 min vs. control)-(relative reperfusion). Limbs reperfused after 6 h ischaemia demonstrated a subsequent decline in muscle blood flow between 120 min and 240 min following revascularisation (p less than 0.05). In addition, muscle blood flow at 240 min was no different to that in a totally ischaemic limb (p less than 0.01 vs. controls; ns vs. ischaemic limb) thus representing reperfusion injury.     

Pre-ischaemic administration of procaine suppresses ischaemic glutamate release and reduces neuronal damage in the gerbil hippocampus

The effects of intracerebroventricular administration of procaine 2 microgramsmol on ischaemic release of neurotransmitter amino acids in the gerbil hippocampal CAI region were investigated using a microdialysis- high-performance liquid chromatography procedure. Histological outcome was examined by comparing delayed neuronal death between animals treated with procaine before ischaemia and animals treated after ischaemia. Transient forebrain ischaemia for 3 min produced increases in dialysate amino acid concentrations. Aspartate, glutamate and glycine reached 331%, 394% and 233% of pre-ischaemic values. Basal concentrations were restored immediately by reperfusion. Pre-ischaemic administration of procaine suppressed peak release and improved histological outcome. However, post-ischaemic administration did not protect against ischaemic neuronal death. Improvement in ischaemic neuronal damage by pre-ischaemic administration of procaine may be related to suppression of excitatory amino acid release.      

Localization of renal oxidative stress and inflammatory response after lithotripsy

OBJECTIVE

To determine if the acute renal oxidative stress and inflammation after extracorporeal shock wave lithotripsy (ESWL), thought to be mediated by ischaemia, is most severe in the portion of the kidney within the focal zone of the lithotripter, and if these effects result primarily from ischaemic injury.

MATERIALS AND METHODS

Pigs (7–8‐weeks old) received either 2000 shock waves at 24 kV to the lower‐pole calyx of one kidney or unilateral renal ischaemia for 1 h. A third group (sham) received no treatment. Timed urine and blood samples were taken for analysis of lipid peroxidation and the inflammatory cytokines, tumour necrosis factor‐α (TNF‐α) and interleukin‐6 (IL‐6). At 4 h after treatment, kidneys were removed and samples of cortex and medulla were frozen for analysis of cytokines and heme oxygenase‐1 (HO‐1).

RESULTS

ESWL did not affect urinary excretion of malondialdehyde, but did elicit an eight‐fold induction of HO‐1 in the portion of the renal medulla within the focal zone of the lithotripter (F2), while remaining unchanged elsewhere in the treated kidney. There was no induction of HO‐1 in renal tissue after ischaemia‐reperfusion. Urinary excretion of TNF‐α increased from the lithotripsy‐treated kidney by 1 h after treatment, but was unaffected by ischaemia‐reperfusion. As with the HO‐1 response after lithotripsy, IL‐6 increased only in the renal medulla at F2. By contrast, ischaemia‐reperfusion increased IL‐6 in all samples from the treated kidney.

CONCLUSION

These findings show that the acute oxidative stress and inflammatory responses to ESWL are localized to the renal medulla at F2. Furthermore, the differing patterns of markers of injury for ESWL and ischaemia‐reperfusion suggest that ischaemia is not the principal cause of the injury response after ESWL.     

Effect of Nifedipine in the Protection of Renal Function During In Situ Preservation

In an experimental study on six healthy dogs both kidneys wereexposed and subjected to 1 h of ischaemia by clamping both renalvessels. To the left renal artery, 300 ml of cold (4°C)Euro-Collins solution in which nifedipine (Bay a 1040—20µg/kg per min) was diluted, was infused for 15 min. Simultaneously300ml of cold Euro-Collins solution plus 20 µg/kg permin of placebo (Bay a 1040—placebo) was infused in theright renal artery. The 1 h of ischaemia was divided in a 15-minperiod of cold ischaemia and 45 min of warm ischaemia, at theend of which both clamps were removed. During the 2 h (4x30min) following the removal of the clamps, urine volume, ureaand creatinine clearances, urine sodium concentration, sodiumfractional excretion (%) and urine/plasma osmolality ratio measurementswere made and results compared to the pre-ischaemia values.Both kidneys were then removed for histological study. The nifedipine group restored diuresis of 0.43±0.23 ml/minwithin 30 min, while this degree of diuresis (0.64±0.16ml/min) was achieved by the placebo group at 120 min. Urinevolume as well as creatinine and urea clearances of the nifedipinegroup were significantly higher in all studied periods comparedto the placebo group (P<0.01 or P<0.025). Urine sodiumand FE_Na (%) were not different between the two groups, andurine/plasma osmolality ratio was above 1.1 in all studied periodsfor both groups. The microscopic study did not show any significantdifferences between the two groups. We conclude that nifedipineis effective in the protection of renal function when it isadministered during experimental in situ preservation.     

Early renal post-ischaemic tissue damage and dysfunction with contribution of A1-adenosine receptor activation in rat

Aim:   This study investigated the effect of a selective A₁-adenosine receptor (A₁-AR) antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), on the renal dysfunction and histological damage induced by ischaemia/reperfusion at an early stage.
Methods:   Pentobarbital anaesthetised rats were prepared for measuring renal functional variables. Ischaemia was induced by bilateral renal artery clamping for 30 min followed by a 4 h reperfusion period. In DPCPX-treated rats, it was infused (i.v.) at 10 µg/kg per min before and after renal ischaemia. Both kidneys were examined using light and electron microscopy.
Results:   The renal ischaemic challenge resulted in major histological and ultrastructural damages, which were associated with decreased creatinine clearance, absolute potassium-excretion and effective free-water reabsorption, but increased fractional sodium-excretion and urine flow during reperfusion period. In DPCPX-treated rats, the histological and ultrastructural damage to the kidneys was improved along with the decrease in creatinine clearance and increase in fractional sodium-excretion being smaller, but the increase in urine flow being larger than those of the non-treated rats, while absolute potassium-excretion and effective free-water reabsorption were equal to those of the sham-operated rats.
Conclusion:   These findings suggest that endogenous activation of A₁-AR contributes to the early development of renal ischaemia/reperfusion injury.     

Effects of the free radical scavenger dimethyl sulphoxide on experimental normothermic ischaemia of the liver

BACKGROUND/AIMS: This study assessed the effects of intermittent or continuous hepatic ischaemia and reperfusion with or without dimethyl sulphoxide (DMSO) pre-treatment in a rat ischaemic model. METHODS: One hundred and eighty rats were divided into three groups undergoing hepatic ischaemia of a total duration of 60, 90 and 120 min. Each group of rats was subdivided to receive either a continuous Pringle manoeuvre or intermittent liver pedicle clamping of 30 or 15 min. Ten minutes before ischaemia induction, 10 rats from each group were pre-treated with DMSO (500 mg/kg, b.w.) intravenously. RESULTS: With continuous hepatic pedicle clamping, survival rates inversely correlated with the duration of ischaemia, with greater survival in the intermittently clamped groups (p < 0.05). DMSO pre-treatment did not affect survival but resulted in a significant reduction in liver enzyme (aspartate aminotransferase, alanine aminotransferase) release on the first postoperative day following total ischaemic times of 90 min or greater (p < 0.05). After 120 min of total ischaemia, DMSO pre-treatment resulted in higher preservation of adenosine 5'-triphosphate liver content (p < 0.05). CONCLUSION: DMSO may be used to prolong tolerance to inflow occlusion and to limit the adverse effects of ischaemia and reperfusion cycles in an experimental hepatic ischaemia model.     

Mesna: a novel renoprotective antioxidant in ischaemic acute renal failure.

BACKGROUND: Reactive oxygen species (ROS) play a key role in renal ischaemia-reperfusion injury. After establishing the in vitro anti-oxidative potential of mesna, a sulfhydryl-containing compound, its effect on kidney function and morphology in a rat model of ischaemic acute renal failure (ARF) was examined. METHODS: Mesna (180 mg/kg) was administered at different time points relative to ischaemia and/or reperfusion onset. Kidney function was assessed by glomerular filtration rate (GFR) and fractional sodium excretion (FE(Na)) before a 45-min period of unilateral renal artery clamping and following 90 min of reperfusion. Mesna was administered by bolus, 30 min before the induction of ischaemia, 5 min before ischaemia, 5 min before reperfusion, and 5 min after the onset of reperfusion. RESULTS: Mesna improved function of the ischaemic kidney at each administration. When mesna was administered 5 min before the onset of reperfusion, GFR reached 90-100% of its pre ischaemic value and FE(Na) was improved by 75%. The beneficial effect of mesna was also demonstrated by light and electron microscopy. Kidneys treated with mesna 5 min before reperfusion resembled ischaemic non-reperfused kidneys and showed subtle morphological and ultrastructural changes compared with ischaemic-reperfused kidneys. Mesna had no haemodynamic effect on renal blood flow and did not induce any osmotic diuresis. CONCLUSIONS: We suggest that mesna acts as an antioxidant. Its antioxidant potential together with optimal protection achieved when administered 5 min before reperfusion, supports the conclusion that mesna scavenges ROS generated at the onset of reperfusion, thus diminishing reperfusion injury and organ damage.     

Attenuation of reperfusion injury by renal ischaemic preconditioning: the role of nitric oxide

OBJECTIVE: To determine the effect on nitric oxide (NO) release and renal NO synthase (endothelial, eNOS and inducible, iNOS) activity of renal ischaemia-reperfusion (I/R) in vivo in an animal model, and to examine the possible involvement of NO in ischaemic preconditioning (IP) of the kidney. MATERIALS AND METHODS: In a right-nephrectomized rat model, 42 animals were randomized in four groups: controls; IP-only (4 min of ischaemia followed by 11 min of reperfusion, total of four cycles); renal warm ischaemia (45 min) and 6 h reperfusion; ischaemia (45 min) preceded by IP pretreatment. Serum NO metabolites were assayed 2 and 6 h after ischaemia or the control equivalent. NOS expression in the kidney was detected immuno-histochemically, and damage assessed morphologically in sections stained with haematoxylin and eosin. Kidney function was assessed by the levels of serum creatinine, urea and electrolytes. RESULTS: Compared with before ischaemia, the concentration of serum NO metabolites at 6 h was increased in the IP-only animals (P = 0. 016) and in the IP + I/R group (P = 0.002). There was greater eNOS expression in the IP-only group (P = 0.009) and in the IP + I/R group than in controls (P = 0.050). iNOS expression was greater in the IP-only animals than in the control group (P = 0.050). Histological assessment showed less evidence of cellular damage in IP + I/R animals than in the I/R-alone group (P = 0.020). Serum creatinine level was not significantly different between the IP-only group and the control. There were no differences after 2 h of reperfusion. CONCLUSION: Ischaemic preconditioning has a protective effect on renal structure and function, which may be produced by increased NO release arising from increased NOS expression by 6 h after reperfusion.     

The Effect of Diosmin Hesperidin on Intestinal Ischaemia — Reperfusion Injury

Aim: The effect of Diosmin Hesperidin on intestinal ischaemia reperfusion injury was evaluated in an experimental model in rats.

Material and methods: Forty Spraque-Dawley rats were divided into 4 groups of (n = 10) (sham, sham + Diosmin Hesperidin, Reperfusion, Reperfusion + Diosmin Hesperidin). Diosmin Hesperidin oral gavage was administrated at a dose of 50 mg/kg to rats 14 and 2 hours before the operation and 30 minutes of ischaemia and 30 minutes of reperfusion was performed in the groups when appropriate. Ileum samples were resected for histopathological evaluation and tissue malondialdehyde (MDA) and myeloperoxidase (MPA) level determination.

Results: Mean mucosal injury score of IR group (4,50 ± 0,23) was significantly higher than the other groups (p < 0.05). Although mean mucosal injury score of IR + DH group was higher than sham and sham + DH groups, difference was not statistically significant (p > 0.05). Tissue MDA and MPO activities of IR group were 45,55 ± 2.61 nmol/g/wet tissue and 1.68 ± 0.25 U/g/wet tissue respectively and were significantly higher than the other groups (p < 0.008). Although tissue MDA and MPO activities of IR + DH group was higher than sham and sham + DH groups, differences were not statistically significant (p > 0.008).

Conclusion: Diosmin Hesperidin seems to be effective in the prevention of intestinal reperfusion injury.     

-Arginine given after ischaemic preconditioning can enhance cardioprotection in isolated rat hearts

Objective: Ischaemic or pharmacological preconditioning with -arginine has been reported to be insufficient for optimal cardioprotection. The ability of nitric oxide (NO) to enhance ischaemic preconditioning was assessed, and the role of -arginine-induced ischaemic preconditioning in myocardial protection was determined. Methods: Isolated rat hearts were prepared and divided into six groups: control hearts (control, n=6) were perfused without global ischaemia at 37°C for 160 min; global ischaemia hearts (GI, n=6) were subjected to ischaemia for 20 min and reperfusion for 120 min; ischaemic preconditioned hearts (IP, n=6) received 2 min of zero-flow global ischaemia followed by 5 min reperfusion, before 20 min of global ischaemia; -arginine hearts (ARG, n=6) received 1 mmol/l -arginine for 5 min, before 20 min of global ischaemia; ischaemic preconditioning plus nitro- -arginine methyl ester hearts (IP+ -NAME, n=6) received 2 min of ischaemic preconditioning and 5 min reperfusion with 3 mmol/l -NAME in Krebs–Henseleit buffer, before 20 min of global ischaemia; and ischaemic preconditioning plus -arginine hearts (IP+ARG, n=6) received 2 min of ischaemic preconditioning and 5 min reperfusion with 1 mmol/l -arginine in Krebs–Henseleit buffer. Haemodynamic parameters and coronary flow were recorded continuously. Nitrites and nitrates (NOx) were measured 5 and 60 min after reperfusion, and infarct size was also determined. Results: In the IP+ARG group, significant amelioration and preservation of left ventricular peak developed pressure and coronary flow was observed compared with the GI, IP, ARG and IP+ -NAME groups. Infarct size in the IP+ARG group was reduced significantly compared with that in the GI, IP, ARG and IP+ -NAME groups. Significant preservation of NOx was observed during reperfusion in the IP+ARG group compared with the GI group. Conclusions: Inhibition of NO synthase with -NAME had little impact on ischaemic preconditioning, suggesting that endogenous NO is not a major mediator of ischaemic preconditioning. Nevertheless, enhancement of the effects of ischaemic preconditioning can be achieved with -arginine, a precursor of NO, improving post-ischaemic functional recovery and infarct size in the isolated rat heart.     

Protective effect of UR-12670 on chronic nephropathy induced by warm ischaemia in ageing uninephrectomized rats.

BACKGROUND: In young animals, renal ischaemia/reperfusion injury and mass reduction are associated with chronic lesions that mimic those found in chronic rejection. We have shown that the phospholipid platelet-activating factor (PAF) participates in young animals in such chronic nephropathy. Here we examine the long-term effects of the orally active PAF antagonist, UR-12670 in ageing uninephrectomized rats exposed to prolonged warm ischaemia. METHODS: Fifteen- to eighteen-month-old uninephrectomized male Sprague-Dawley rats were allocated into three groups and followed for 16 weeks: UNx, rats without ischaemia; UNxISC, ischaemic kidney (60 min), and UNxISC+UR, ischaemic kidney and UR-12670 from day 0 to the 16th week. Serum creatinine and proteinuria were monitored every 4 weeks. At the end of the study, conventional histology was performed and monocyte-macrophages were identified with the specific monoclonal antibody ED-1. RESULTS: The UNxISC group had severe acute renal failure with a high mortality rate, which was associated with incomplete restoration of renal function. Renal insufficiency in this group was sustained throughout the follow-up. Both UNx and UNxISC groups developed progressive proteinuria from the 12th week. Though UNxISC+UR group showed similar acute renal failure and mortality rate to the ischaemic non-treated group, serum creatinine decreased to levels similar to UNx group, which were maintained until the end of the study. Treatment of ischaemic kidneys with UR-12670 produced a slight decrease in 24-h proteinuria and a reduction in glomerulosclerosis, the mean tubulointerstitial score and number of monocyte-macrophages to values similar to UNx group. CONCLUSIONS: The chronic administration of the PAF antagonist UR-12670 attenuates the long-term effects of ischaemia-reperfusion injury in uninephrectomized ageing rats. The beneficial effect of this agent suggests that PAF contributes to the progression to late renal damage in this model.     

Effect of bencyclane fumarate on intestinal ischaemia reperfusion injury

Background: Post‐ischaemic intestinal tissue damage appears to be due to the formation of oxygen radicals. Free radical‐initiated lipid peroxidation following intestinal ischaemia/reperfusion (I/R) may disrupt mucosal integrity. Indirectly, the radicals trigger the accumulation of neutrophils within the affected tissue, initiating inflammatory processes that lead to severe mucosal lesions. We have investigated the protective effect of bencyclane fumarate, a vasodilating Ca²⁺ channel blocker, which has been used for the treatment of peripheral arterial occlusive diseases, on intestinal ischaemia reperfusion (IR) injury in rats. Methods: Forty‐eight Wistar albino rats were divided into three groups: a sham‐operated group (no IR injury, n = 16), an ischaemic control group (IR, n = 16), and BF‐treated group (pretreatment 5 mg/kg bencyclane fumarate + IR, n = 16). A marker for lipid peroxidation, namely malondialdehyde; free radical scavengers, glutathione peroxidase, catalase and superoxide dismutase levels; an index of polymorphonuclear neutrophils, myeloperoxidase activity and mucosal damage were investigated. Results: Malondialdehyde levels, myeloperoxidase activity and the severity of mucosal damage were decreased in the BF group. In addition, in the BF group, glutathione peroxidase, catalase and superoxide dismutase levels were higher compared with the IR group. Conclusion: The pretreatment of rats with bencyclane fumarate before intestinal ischaemia attenuates the mucosal damage in intestinal IR injury, probably by altering lipid peroxidation, neutrophil accumulation and antioxidant activity.     

Role of combination of L-arginine and α-tocopherol in renal transplantation ischaemia/reperfusion injury: a randomized controlled experimental study in a rat model

What’s known on the subject? and What does the study add? Renal ischaemia/reperfusion (I/R) injury is an inevitable consequence of kidney transplantation. It contributes to delayed graft function (DGF), acute renal failure and graft rejection. The present study investigates for the first time the impact of a combination of l ‐arginine and alpha tocopherol on the renal ischemia/reperfusion injury in a rodent model of kidney transplation. We found that concomitant administration of l ‐arginine and α‐tocopherol has a more protective effect and synergistic antioxidant effect on ischaemia/reperfusion injury in transplanted rat kidneys.

OBJECTIVES

To investigate the role of l ‐arginine and α‐tocopherol in ischaemia/reperfusion injury in a kidney transplanted rat model.

MATERIALS AND METHODS

In total, 40 male Sprague‐Dawley rats subjected to renal transplantation received FK506 (tacrolimus) to overcome early acute rejection episodes. Animals were divided randomly into four groups (ten rats each). Group I were treated with FK506 (2 mg/kg/bw/day) and served as the control group. Group II were treated with l ‐arginine 300 mg/kg/bw. Group III were treated with α‐tocopherol 30 mg/kg/bw. Group IV were treated with l ‐arginine and α‐tocopherol. Urine and blood samples were taken at 0 (before operation), 2, 7 and 14 days post‐transplantation for estimation of urine sodium, creatinine, fractional excretion of sodium, serum creatinine, sodium and blood urea nitrogen. Histological examination and measurement of malondialdehyde in kidney tissues were also performed.

RESULTS

Serum creatinine and blood urea nitrogen significantly decreased in l ‐arginine and α‐tocopherol, as well as combination groups, compared to the control group. Malondialdehyde was significantly decreased in the combination group compared to l ‐arginine and α‐tocopherol alone. Histological examination of the control group showed that acute tubular necrosis was markedly decreased in transplanted kidneys treated with a combination of both l ‐arginine and α‐tocopherol.

CONCLUSIONS

Concomitant administration of l ‐arginine and α‐tocopherol has a more protective effect and synergistic antioxidant effect on ischaemia/reperfusion injury in transplanted rat kidneys.     

Remote ischaemic preconditioning of the hind limb reduces experimental liver warm ischaemia-reperfusion injury

BACKGROUND: Direct ischaemic preconditioning of the liver reduces ischaemia-reperfusion injury (IRI). Remote ischaemic preconditioning (RIPC) of a limb has been shown to reduce IRI to the heart. This study determined the effect of brief remote ischaemia to the limb in reducing early liver warm IRI. METHODS: Twenty-eight male rabbits were allocated to four groups: sham operated, RIPC alone, IRI alone, and RIPC plus IRI. RIPC was induced in the leg with a tourniquet, before liver IRI, by three alternate cycles of 10 min ischaemia followed by 10 min reperfusion. Liver IRI was produced by total inflow occlusion for 25 min. Markers of liver injury and systemic and hepatic haemodynamics were measured for 2 h after reperfusion. RESULTS: At 2 h, IRI alone was associated with increased serum levels of aminotransferases, and reduced mean arterial blood pressure, hepatic blood flow and peripheral oxygen saturation. There was significant improvement in these variables in animals that had RIPC before liver IRI, and hepatic venous nitrate/nitrite levels were also significantly higher. CONCLUSION: In this experimental model RIPC appeared to reduce liver IRI.     

Isoflurane-induced myocardial preconditioning is dependent on phosphatidylinositol-3-kinase/Akt signalling

Background. Isoflurane and other volatile anaesthetics havea cardioprotective effect and limit myocardial infarct sizeto the same extent as ischaemic preconditioning. Phosphatidylinositol-3-kinase(PI3K) was found to play a key role in myocardial protectionby ischaemic preconditioning. The aim of the present investigationwas to evaluate whether isoflurane-induced myocardial preconditioningis dependent on PI3K signalling. Methods. Using a model of regional myocardial ischaemia andreperfusion, New Zealand White rabbits were subjected to 40min of regional myocardial ischaemia followed by 120 min ofreperfusion. The rabbits were randomly assigned to one of thefollowing six experimental groups: sham-operated controls (n=5);ischaemia and reperfusion controls (n=8); isoflurane preconditioning(n=8); a PI3K inhibitor, wortmannin (0.6 mg kg^–1 i.v.)+ isoflurane (n=8); and wortmannin+ischaemia and reperfusion(n=8). An additional control group of sham operation+ wortmannin(n=5) was also included. Myocardial injury was assessed by measuringthe serum concentration of the MB fraction of creatine kinase(CK-MB) and infarct size was assessed by 2,3,5-triphenyl tetrazoliumchloride staining. Phosphorylation of Akt, a downstream targetof PI3K, was assessed by western blotting. Results. Isoflurane preconditioning was seen as reduced infarctsize compared with control animals: 24 (4) and 41 (5)% respectively(P<0.05). Wortmannin inhibited this cardioprotective effectwith myocardial infarct size at 44 (3)% (not significant). Aktphosphorylation was increased after isoflurane preconditioning,but administration of wortmannin blocked this effect. Conclusions. Our data demonstrate that isoflurane protects theheart against ischaemia and decreases myocardial infarctionby activation of PI3K.