Role of the L-arginine/nitric oxide pathway in renal ischaemia-reperfusion in rats.

OBJECTIVE: To study the role of the L-arginine/nitric oxide (NO) pathway during renal ischaemia-reperfusion in rats. DESIGN: Randomised experimental study. SETTING: Teaching hospital, Brazil. ANIMALS: 97 male Wistar rats randomly assigned to 4 groups for the assessment of renal dysfunction and to 6 groups for the assessment of the oxidative stress induced on renal cell membranes by ischaemia-reperfusion. INTERVENTIONS: The animals underwent sham-operation or renal ischaemia-reperfusion (n = 9 each) with or without pretreatment with L-arginine (a NO donor) or L-NAME (N(omega)-nitro-L-arginine methyl ester--an inhibitor of NO production) (n = 10 each). MAIN OUTCOME MEASURES: Serum creatinine concentrations and oxidative stress by chemiluminescence initiated by the tert-butyl hydroperoxide technique. RESULTS: Renal ischaemia-reperfusion significantly worsened renal dysfunction and increased oxidative stress in the ischaemia-reperfusion group after 24 and 96 hours of reperfusion compared with the control group (p < 0.05). Pretreatment with L-NAME slightly but not significantly increased serum creatinine concentrations after 24 and 96 hours of reperfusion together with activity of reactive oxygen species during renal ischaemia-reperfusion. L-arginine also significantly protected renal function and reduced the increment in the amount of chemiluminescence induced by giving L-NAME during 24 and 96 hours of reperfusion (p < 0.05). CONCLUSION: The L-arginine/NO pathway seems to have a slightly protective effect on the kidney after renal ischaemia-reperfusion injury in rats. These results need to be confirmed by studies in human beings.     

Leucocyte depletion improves renal function during reperfusion using an experimental isolated haemoperfused organ preservation system

BACKGROUND: Leucocytes have been implicated as mediators of renal ischaemia-reperfusion injury. This study aimed to demonstrate the effect of white cells in early renal reperfusion injury using an isolated haemoperfused porcine kidney model. METHODS: After 2 h cold storage, porcine kidneys were perfused with normothermic autologous blood using an isolated organ preservation system. This was designed using cardiopulmonary bypass technology, and perfusion commenced with a circulating serum creatinine level of 1000 micromol/l. In group 1 (n = 6) a leucocyte filter was included in the circuit and in group 2 (n = 6) non-filtered blood was used. RESULTS: The mean(s.d.) area under the curve for serum creatinine was lower in the leucocyte-depleted experiments (1286(214) versus 2627(418); P = 0.002). Leucocyte depletion also led to improved urine output (191(75) versus 70(32) ml/h; P = 0.002) and higher creatinine clearance (10.6(2.8) versus 1.9(1.0) ml/min; P = 0.002). Renal blood flow, oxygen consumption and acid-base homeostasis were all improved by perfusion with leucocyte-depleted blood, and histological tubular damage was ameliorated. CONCLUSION: These data show that the depletion of leucocytes from blood used to perfuse porcine kidneys improved postschaemic renal function, indicating that white cells play an important role in renal ischaemia-reperfusion injury.     

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.     

Cardioprotection by breathing hyperoxic gas—relation to oxygen concentration and exposure time in rats and mice

Objectives: Breathing a hyperoxic gas (≥95% O₂) protects against ischaemia-reperfusion injury in rat and mouse hearts. The present study investigated how oxygen concentration and duration of hyperoxic exposure influenced cardioprotection, and whether hyperoxia might induce delayed cardioprotection (after 24 h). Methods: Animals were kept in normal air or in a hyperoxic environment, and their hearts were isolated and Langendorff-perfused immediately or 24 h thereafter. Global ischaemia was induced for 25 min in rats and 40 min in mice, followed by 60 min of reperfusion. Infarct size was determined by triphenyl tetrazolium chloride staining. Results: In rats exposure to ≥95, 80, and 60%, but not to 40% of oxygen immediately before heart isolation and perfusion improved postischaemic functional recovery. Eighty or more percent of oxygen also reduced infarct size. A preconditioning-like effect could be evoked by 60 or 180 min of hyperoxia, giving both immediate and delayed protection. In the mouse heart protection could be induced by pretreatment for 15 or 30, but not by 60 min with ≥95% oxygen. The protective effect of hyperoxia in mice could be evoked in the immediate model only. Conclusions: Hyperoxia protects the isolated rat and mouse heart against ischaemia-reperfusion injury, but some species-different responses exist. The protection depends on both oxygen concentration in inspired air, and duration of hyperoxic exposure.     

Do prostaglandins have a salutary role in skeletal muscle ischaemia-reperfusion injury?

OBJECTIVES: the effects of prostaglandins (PG) E1, E2, and the prostacyclin analogue iloprost with and without the addition of free-radical scavengers catalase and superoxide dismutase on gastrocnemius blood flow and oedema were studied in a rodent model of hindlimb ischaemia-reperfusion. METHODS: male Sprague-Dawley rats underwent 6-h hindlimb ischaemia with 4-h reperfusion. Prostaglandins were infused prior to reperfusion and their effects on limb blood flow and oedema examined. RESULTS: control animals exhibited a triphasic pattern of muscle blood flow during reperfusion compared to normal animals. PGE1 did not abolish low reflow at 10 min, relative reperfusion was preserved but reperfusion injury was abolished at 120 min. Muscle blood flow was increased at 240 min compared to controls. Increased limb swelling was also seen. Addition of free-radical scavengers caused the abolition of low reflow. Similar results were seen with iloprost. PGE2 abolished low reflow at 10 min and increased perfusion at 120 min but did not prevent reperfusion injury at 240 min. CONCLUSIONS: PGE1 and iloprost enhance muscle blood flow at 4-h reperfusion, though neither abolishes low reflow; PGE2 improved flow at 10 and 120 min but not after 240 min. This study demonstrates a potentially beneficial role for prostaglandins in improving muscle blood flow in skeletal muscle ischaemia-reperfusion injury.     

Pre-treatment with hyperoxia before coronary artery bypass grafting - effects on myocardial injury and inflammatory response

BACKGROUND: In experimental studies, exposure to hyperoxia for a limited time before ischaemia induces a low-grade systemic oxidative stress and evokes an (ischaemic) preconditioning-like effect of the myocardium. We hypothesised that hyperoxia before cardioplegia could protect the myocardium against necrosis and stunning caused by ischaemia-reperfusion. METHODS: Forty patients undergoing coronary artery bypass grafting were randomly exposed to an oxygen fraction of 0.4 or > 0.96 in inspired air on an average of 120 min before cardioplegia. Blood for troponin I, creatine kinase-MB, lactate, glutathione and interleukin-6 was sampled from arterial and coronary sinus cannulae during 20 min of reperfusion. Additional arterial samples were drawn 60 min after declamping and in the first post-operative morning. The cardiac index and right and left ventricular stroke work indices were measured before sternotomy and up to 12 h post-operatively. RESULTS: Troponin I, creatine kinase-MB and lactate did not differ between the groups. Hyperoxic pre-treatment had no impact on the post-operative haemodynamic indices measured with the thermodilution pulmonary artery catheter. More oxidised glutathione was released in the hyperoxia group in the first minute of reperfusion (P = 0.015). Hyperoxic pre-treatment abolished the myocardial release of interleukin-6 during 20 min of reperfusion (P = 0.021 vs. controls). In the first post-operative morning, interleukin-6 was higher in the hyperoxia group [127.0 (86.0-140.0) vs. 85.2 pg/ml (66.6-94.5 pg/ml); P = 0.016]. Conclusions: Exposure to >96% oxygen before cardioplegia did not attenuate ischaemia-reperfusion injury of the heart in patients undergoing coronary artery bypass grafting. The only potentially beneficial effect observed was the decreased transmyocardial release of interleukin-6.     

Effects of halothane,isoflurane and sevoflurane on ischemiareperfusion injury in the perfused liver of fasted rats

Background : Although intraoperative ischemia-reperfusion of the liver generally occurs under general anesthesia, little is known about the direct effect of anesthetic agents on hepatic injury due to this phenomenon. The effect of volatile anesthetics on ischemia-reperfusion injury was studied using isolated liver perfusion. Methods : The liver was isolated from 24-h-fasted male Sprague-Dawley rats and perfused through the portal vein with a modified Krebs-Ringer bicarbonate solution in a recirculating perfusion-aeration system. Ischemia was induced by reducing the baseline perfusion pressure from 1.2 to 0.2 kPa followed by reperfusion to baseline level. The ischemia-reperfusion injury was assessed by LDH release from the perfused liver. We studied the effect of halothane, isoflurane and sevoflurane on the ischemia-reperfusion injury during 20 min of control conditions, exposure of the liver to 60 min of ischemia and reperfusion for 90 min. Results : Ischemia was evident by reduced portal vein flow and oxygen consumption, and caused an increase in lactate production. Reperfusion caused a transient reduction in lactate production and a significant increase in LDH release. All anesthetics reduced hepatic oxygen consumption and increased the net lactate production during control conditions. Volatile anesthetics also significantly attenuated LDH release during reperfusion. The suppression of LDH release was observed even when isoflurane was administered during the reperfusion period, but not when it was administered only during ischemia. Conclusion : These results indicate that volatile anesthetics may protect the fasted liver from early, neutrophil-independent, ischemia-reperfusion injury by acting during the reperfusion phase.     

Mice lacking the 110-kD isoform of poly(ADP-ribose) glycohydrolase are protected against renal ischemia/reperfusion injury

The role of poly(ADP-ribose) (PAR) glycohydrolase (PARG) in the pathophysiology of renal ischemia/reperfusion (I/R) injury is not known. Poly(ADP-ribosyl)ation is rapidly stimulated in cells after DNA damage caused by the generation of reactive oxygen and nitrogen species during I/R. Continuous or excessive activation of poly(ADP-ribose) polymerase-1 produces extended chains of ADP-ribose on nuclear proteins and results in a substantial depletion of intracellular NAD(+) and subsequently, ATP, leading to cellular dysfunction and, ultimately, cell death. The key enzyme involved in polymer turnover is PARG, which possesses mainly exoglycosidase activity but can remove olig(ADP-ribose) fragments via endoglycosidic cleavage. Thus, the aim of this study was to investigate whether the absence of PARG(110) reduced the renal dysfunction, injury, and inflammation caused by I/R of the mouse kidney. Here, the renal dysfunction and injury caused by I/R (bilateral renal artery occlusion [30 min] followed by reperfusion [24 h]) in mice lacking PARG(110), the major nuclear isoform of PARG, was investigated. The following markers of renal dysfunction and injury were measured: Plasma urea, creatinine, aspartate aminotransferase, and histology. The following markers of inflammation were also measured: Myeloperoxidase activity, malondialdehyde levels, and plasma nitrite/nitrate. The degree of renal injury and dysfunction caused by I/R was significantly reduced in PARG(110)-deficient mice when compared with their wild-type littermates, and there were no differences in any of the biochemical parameters measured between sham-operated PARG(110)(-/-) mice and sham-operated wild-type littermates. Thus, it is proposed that endogenous PARG(110) plays a pivotal role in the pathophysiology of I/R injury of the kidney.     

Enhanced post-ischaemic recovery in rabbit kidney after pretreatment with an indeno-indole compound and ascorbate monitored in vivo by 31P magnetic resonance spectroscopy

OBJECTIVE: To investigate whether combined pretreatment with lipid- and water-soluble antioxidants gave better restoration of energy phosphates after ischaemia-reperfusion of rabbit kidneys than single pretreatment with a lipid-soluble antioxidant. MATERIAL AND METHODS: Thirteen New Zealand white rabbits were used for the study. Changes in energy phosphates were measured in vivo using volume-selective 31P magnetic resonance spectroscopy. The indeno-indole compound H290/51 was chosen as a lipid-soluble antioxidant and ascorbate as a water-soluble antioxidant. RESULTS: The combined pretreatment led to significantly better restoration of the beta-adenosine triphosphate:inorganic phosphate ratio after 60 min of ischaemia and 120 min of reperfusion compared with the single pretreatment. Analyses of blood pressure and blood gas changes showed that the beneficial effect of combined pretreatment was not caused by a better general condition of the animals in that group but by a direct effect on the kidneys. CONCLUSIONS: Combined pretreatment with lipid- and water-soluble antioxidants leads to better restoration of energy phosphates in rabbit kidneys subjected to ischaemia-reperfusion compared with single pretreatment with a lipid-soluble antioxidant.     

利多卡因预先给药对肾脏缺血再灌注损伤大鼠肾组织CD44和TNF-α表达的影响

目的 评价利多卡因预先给药对肾脏缺血再灌注损伤大鼠肾组织CD44和TNF-α表达的影响.方法 健康雄性Wistar大鼠36只,体重300～350 g,随机分为3组(n=12):假手术组(S组)、肾脏缺血再灌注组(IR组)和利多卡因预先给药组(L组).采用无创动脉夹夹闭双侧肾动脉60min、恢复灌注4 h,建立大鼠肾脏缺血再灌注模型.L组于夹闭双侧肾动脉前5min时尾静脉注射利多卡因5 mg/kg;IR组于夹闭双侧肾动脉前5 min时尾静脉注射等容量生理盐水;S组不夹闭双侧肾动脉,于分离肾动脉后尾静脉注射等容量生理盐水.再灌注4 h时处死大鼠,取肾组织,光镜下观察病理学结果;采用免疫组化法测定肾组织CD44和TNF-α的表达水平.结果 与S组比较,IR组肾组织CD44和TNF-α的表达上调(P＜0.05),L组肾组织CD44和TNF-α的表达差异无统计学意义(P＞0.05);与IR组比较,L组肾组织CD44和TNF-α的表达下调(P＜0.05),肾组织损伤减轻.结论 利多卡因预先给药减轻大鼠肾脏缺血再灌注损伤与其抑制肾组织CD44和TNF-α的表达有关.     

Effect of low-dose N-acetyl-cysteine infusion on tourniquet-induced ischaemia-reperfusion injury in arthroscopic knee surgery

BACKGROUND: Temporary occlusion of blood flow is used during arthroscopic knee surgery in order to provide a bloodless surgical field. The resulting ischaemia-reperfusion causes lipid peroxidation, which contributes to tissue injury. The aim of the study was to investigate the effect of low-dose n-acetyl cysteine (NAC) infusion on oxidative stress by determining malondialdehyde (MDA) levels during arthroscopic knee surgery. METHODS: Thirty patients, ASA I - II, undergoing arthroscopic knee debridement under a tourniquet were divided into NAC and control groups. Anaesthesia was induced with propofol, fentanyl and vecuronium bromide and maintained with desflurane in an equal parts O(2)-N(2)O mixture. In the NAC group, an infusion of NAC, 5 mg kg(-1).h(-1), was started after intubation, and continued until extubation. An equal volume of saline was infused to the control group. Duration of ischaemia, anaesthesia time, total dose of NAC infused were also recorded. Venous blood and synovial membrane tissue samples were obtained 10 min after the onset of NAC infusion but before tourniquet inflation (t1), after 30 min of ischaemia (t2), and after 5 min of reperfusion following tourniquet release (t3). RESULTS: Plasma MDA levels were significantly lower in the NAC group on reperfusion. There were no differences between the groups in tissue MDA levels at ischaemia and reperfusion times. CONCLUSION: Low-dose n-acetyl cysteine infusion attenuates lipid peroxidation and ischaemia-reperfusion injury in arthroscopic knee surgery requiring tourniquet application.     

Recombinant soluble P-selectin glycoprotein ligand 1 moderates local and remote injuries following experimental lower-torso ischaemia.

BACKGROUND: A central role for the polymorphonuclear leucocyte (PMN) in skeletal muscle ischaemia-reperfusion has been demonstrated by the observation that PMN depletion reduced local and remote pulmonary vascular permeability. This study investigated the role of recombinant soluble P-selectin glycoprotein ligand-immunoglobulin fusion protein (rPSGL-Ig), a P- and E-selectin antagonist, in moderating injury. METHODS: Mice underwent 2 h of hindlimb ischaemia and 3 h of reperfusion. Muscle and lung vascular permeability index (PI) was assessed by extravasation of (125)I-radiolabelled albumin. Lung myelo peroxidase (MPO) activity was also measured. RESULTS: In mice treated with rPSGL-Ig 1 mg/kg before reperfusion (n = 12) muscle PI was reduced by 40 per cent, whereas it was moderated by 20 per cent in animals treated 30 min after reperfusion (n = 15). Lung PI in mice treated with rPSGL-Ig before (n = 12) and 30 min after (n = 15) reperfusion was reduced by over 99 and 98 per cent respectively. Lung MPO activity in mice treated with rPSGL-Ig before (n = 10) and 30 min after (n = 12) reperfusion was reduced by 68 and 58 per cent respectively. Treatment with rPSGL-Ig 1 h after reperfusion, or with m20ek.Fc 1 mg/kg (n = 9; negative control for rPSGL-Ig which is inactive for selectin binding) before reperfusion failed significantly to moderate local or remote organ injury. CONCLUSION: Selectin blockade moderated local skeletal muscle and remote lung injury following hindlimb ischaemia--reperfusion. Significantly, delayed antiselectin therapy also decreased injury.     

Effects of L-arginine on the kidney levels of malondialdehyde in rats submitted to renal ischaemia-reperfusion

OBJECTIVE: To evaluate the effects of L-arginine, a nitric oxide donor, on kidney levels of malondialdehyde (MDA, a product of cellular lipid peroxidation), serum creatinine levels, and urinary volume in rats undergoing unilateral renal ischaemia-reperfusion. MATERIALS AND METHODS: Wistar rats (117) were randomly distributed into three experimental groups (of four subgroups each) in which were assessed renal cell-lipid peroxidation (kidney levels of MDA), serum creatinine levels and urinary volume. The rats underwent unilateral nephrectomy followed by contralateral renal ischaemia-reperfusion with or with no pretreatment with L-arginine (200 mg/kg) given intraperitoneally. RESULTS: Pretreatment with L-arginine caused significantly higher kidney levels of MDA than in the untreated group (P < 0.05). Furthermore, L-arginine given before surgery attenuated the increase in serum creatinine and significantly increased urinary volume in rats subjected to renal ischaemia-reperfusion (P < 0.05). CONCLUSION: L-arginine tended to be of benefit for renal function during renal ischaemia-reperfusion in rats. Pretreatment with L-arginine (200 mg/kg intraperitoneally) seems to increase the renal damage by increasing kidney levels of MDA.     

Effect of desferrioxamine on reperfusion damage of rat heart mitochondria

Ischaemia of the myocardium leads to necrosis unless oxygen supply is restored but it has only recently been realised that reperfusion is not without danger. The greatest rate of myocardial damage, as measured by mitochondrial function, occurred during the first 5 minutes of reperfusion in rat hearts subjected to normothermic ischaemic cardiac arrest. Addition of desferrioxamine to the perfusate after 5 minutes of reperfusion did not reverse the mitochondrial damage. It is therefore concluded that desferrioxamine prevents mitochondrial damage caused by ischaemia-reperfusion but does not reverse the damage already present.     

Energy metabolism and renal ischemia.

In renal preservation, the longer the organ is cold stored the greater the damage to the organ. The mechanism of hypothermic-induced kidney injury is not known. In this study the effects of long-term preservation (up to 120 h) of the dog kidney on mitochondrial functions in an homogenate of kidney cortex tissue was investigated. Kidneys were exposed to either warm ischemia (0 to 90 min) cold ischemia (0, 72, 96, and 120 h). The mitochondrial oxygen uptake was measured in an homogenate. In both warm and cold ischemia there were changes in the mitochondrial utilization of oxygen. The changes were characterized as a decrease in uncoupler stimulated oxygen uptake by up to 40%, an increase in oligomycin-sensitive respiration by up to about 150%, and a decrease in the respiratory control ratio (uncoupler control ratio) from about 3 to 1. These changes in mitochondrial utilization of oxygen were partially reversed by including albumin in the respiration medium. Albumin binds free fatty acids and these may originate, during ischemia, from the action of phospholipases during ischemia. The changes in mitochondrial oxygen uptake may result from both the loss of membrane-bound phospholipids and the accumulation of free fatty acids. The changes in mitochondrial activity between 72 h (viable kidneys on transplantation) and 96 to 120 h preservation (nonviable kidneys) were not significant. Furthermore, reperfusion of kidneys preserved for 72 to 120 h resulted in a restoration of mitochondrial oxygen uptake to near normal (control) values. Thus, it does not appear that the limitation of successful long-term renal preservation is due to mitochondrial injury caused by cold ischemia.     

Der postischämische Reperfusionsschaden Biochemische und methodische Grundlagen

The term "ischemic reperfusion injury" encompasses all toxic events in a cell that occur during ischemia and subsequent reoxygenation. These reactions have a significant effect, for example, on the rate of organ survival in kidney transplantation. Reactive oxygen intermediates (ROI) play an important role in the process of postischemic reperfusion. The basic mechanisms of generation and detoxification of ROI as well as the possibilities for their registration and quantification under conditions of ischemic reperfusion injury in the rat kidney are demonstrated in this report. A prerequisite to developing cytoprotective strategies is understanding the precise course of these mechanisms to minimize damage caused by ischemia and the subsequent reperfusion, thus retaining the organ's function to the greatest extent.     

Lipoteichoic acid from Staphylococcus aureus reduces renal ischemia/reperfusion injury

BACKGROUND: The aim of this study was to investigate whether in vivo administration of a low, sub-lethal dose of lipoteichoic acid (LTA), a bacterial wall-fragment derived from the Gram-positive bacterium Staphylococcus aureus, protects the kidney against the renal dysfunction and injury caused by ischemia/reperfusion (I/R). METHODS: Male Wistar rats were administered LTA from S. aureus (1 mg/kg, IP). After 24 hours, rats were subjected to bilateral renal ischemia (45 min) followed by reperfusion (6 h). Serum and urinary markers were measured for the assessment of renal function, tubular and reperfusion-injury. Renal sections were used for histological grading of renal injury and for immunohistochemical localization of P-selectin, inducible nitric oxide synthase (iNOS) and nitrotyrosine (indicative of peroxynitrite formation). Kidney myeloperoxidase (MPO) activity and malondialdehyde (MDA) levels were measured for assessment of polymorphonuclear (PMN) cell infiltration and lipid peroxidation, respectively. Nitric oxide (NO) production was determined by measurement of plasma nitrite/nitrate levels. RESULTS: LTA pretreatment significantly reduced renal dysfunction, tubular and reperfusion-injury caused by I/R of the kidney as well as histological evidence of renal injury. LTA also reduced the expression of P-selectin and kidney MPO activity associated with renal I/R. MDA levels were significantly reduced by LTA pretreatment suggesting a reduction in the lipid peroxidation and formation of reactive oxygen species (ROS). LTA pretreatment also markedly reduced both the expression of iNOS and the formation of nitrotyrosine associated with renal I/R. Although LTA significantly reduced plasma nitrite/nitrate levels associated with I/R, nitrite/nitrate levels remained at levels significantly higher than that measured from the plasma obtained from Sham-operated animals. CONCLUSIONS: These data suggest, to our knowledge for the first time, that LTA pretreatment for 24 hours significantly reduces renal I/R injury. We propose that the mechanism of the protective effect involves reduction of the production of NO, ROS and peroxynitrite subsequent to reduced P-selectin and iNOS expression and PMN recruitment. However, although LTA pretreatment resulted in a reduction of iNOS expression and NO production, we hypothesize that the remaining significant levels of NO contribute to the beneficial actions provided by LTA.     

Oxygen free radicals and glutathione in hepatic ischemia/reperfusion injury

Oxygen free radicals have been implicated as mediators of ischemia/reperfusion injury in a variety of organs. We investigated the role of oxidative injury and endogenous hepatic glutathione (GSH) in liver cell injury associated with complete hepatic ischemia and reperfusion. Forty-five minutes of complete hepatic ischemia followed by reperfusion caused an increase in serum GPT and a fall in hepatic GSH but no increase in hepatic lipid peroxidation products. Chemical depletion of hepatic GSH with diethyl maleate did not cause hepatocellular injury but augmented hepatic ischemia/reperfusion-induced SGPT release and promoted lipid peroxidation. Pretreatment with the selective, membrane-permeable oxygen radical scavenger dimethyl sulfoxide protected against the ischemia/reperfusion-induced drop in hepatic GSH but did not reduce SGPT release in normal rats. In rats sensitized to oxidative injury by depletion of endogenous GSH with diethyl maleate the oxygen radical scavenger protected against ischemia/reperfusion-induced lipid peroxidation and reduced the release of SGPT. These findings suggest that the rich hepatic supply with endogenous GSH has a crucial role in the protection against oxygen radical injury following short periods of total hepatic ischemia. Oxygen radical injury only occurs after depletion of these endogenous GSH stores.     

Renal cilia display length alterations following tubular injury and are present early in epithelial repair.

BACKGROUND: Renal cilia are flow sensors that are required for the maintenance of normal kidney architecture. Defects in this organelle are frequently associated with polycystic kidney disease, but the role of renal cilia during acute tubular injury has not been investigated. METHODS: We have analysed the presence and dimensions of renal cilia following renal ischaemia-reperfusion and ureteral obstruction injury in the mouse, and related these results to injury and repair of the renal tubule. The expression of genes encoding cilium-localized proteins was measured following ischaemia-reperfusion injury. RESULTS: Ischaemia-reperfusion injury was demonstrated to affect the length of cilia in the renal tubule and duct. The average length of renal cilia in the proximal tubule decreases 1 day (2.8 +/- 0.4 microm) and 2 days (3.0 +/- 0.2 microm) after injury, as compared to the control uninjured proximal tubule (4.2 +/- 0.3 microm). Later in the injury and repair process at 4 and 7 days, the average length of cilia increases in both the proximal (7 days = 6.2 +/- 0.3 microm) and distal tubule/collecting duct (4 days = 4.4 +/- 0.3 microm; 7 days = 5.5 +/- 0.4 microm; control 2.5 +/- 0.1 microm). The expression level of genes encoding cilium-localized products did not correlate with the increase in cilium length following ischaemia-reperfusion injury. Ureteral obstruction for 8 days also caused lengthening (8 days UUO = 5.8 +/- 0.3 microm; control 2.5 +/- 0.1 microm) of renal cilia in the distal tubule/collecting duct. During the repair process that follows ischaemia-reperfusion injury, cilia were present on the dedifferentiated cells that proliferate and adopt an epithelial phenotype to facilitate the repair of the ischaemic renal tubule. CONCLUSIONS: We propose roles for the renal cilium in responding to changes in the renal environment caused by injury, and in the repair process that re-establishes the epithelial layer of the damaged renal tubule.     

缺氧诱导因子在大鼠肾缺血再灌注损伤中的表达和意义

目的：探讨缺氧诱导因子-1α（HIF-1α）和氧自由基在肾脏缺血再灌注损伤中的表达情况。方法：SD大鼠随机分为假手术组、缺血组、缺血再灌注组、RNAi阴性对照缺血组及HIF-1αRNAi预处理后的缺血组和预处理后缺血再灌注组，利用RT—PCR和western blot技术检测各组大鼠肾脏中HIF-1α和bcl-2的表达情况。检测细胞超氧化物歧化酶（SOD）的活性和丙二醛（MDA）的含量，间接反应氧自由基的生成量。测定各组大鼠肾功能。取各组大鼠肾组织切片并做HE染色。结果：缺血组HIF-1α的表达在5组中最高（P〈0．05）。与缺血组对比，缺血再灌注后HIF-1α和bcl-2的表达降低，同时氧自由基的生成增加（P〈0．05），肾功能明显降低，但经HIF-1αRNAi预处理后上述指标与假手术组相比改变甚微，肾功能得到明显改善。HE染色可见缺血及再灌注组排列紊乱，胞浆萎缩，纹状缘消失，细胞间质增宽，经HIF-1αsiRNA预处理后，接近假手术组水平。结论：HIF-1α的过度表达可能导致氧自由基生成增加，肾功能受损，从而加重缺血再灌注肾脏的损伤，而HIF-1αRNAi可以减轻并改善这一状况，其机制可能与上调凋亡抑制基因有关。