Ischemic preconditioning reduces endoplasmic reticulum stress and upregulates hypoxia inducible factor-1α in ischemic kidney: the role of nitric oxide.

BACKGROUND: Although recent studies indicate that renal ischemic preconditioning (IPC) protects the kidney from ischemia-reperfusion (I/R) injury, the precise protective mechanism remains unclear. In the current study, we investigated whether early IPC could upregulate hypoxia inducible transcription factor-1α (HIF-1α) expression and could reduce endoplasmic reticulum (ER) stress after renal I/R and whether pharmacological inhibition of nitric oxide (NO) production would abolish these protective effects. METHODS: Kidneys of Wistar rats were subjected to 60 min of warm ischemia followed by 120 min of reperfusion (I/R group), or to 2 preceding cycles of 5 min ischemia and 5 min reperfusion (IPC group), or to intravenously injection of NG-nitro-L-arginine methylester (L-NAME, 5 mg/kg) 5 min before IPC (L-NAME+IPC group). The results of these experimental groups were compared to those of a sham-operated group. Sodium reabsorption rate, creatinine clearance, plasma lactate dehydrogenase (LDH) activity, tissues concentrations of malonedialdehyde (MDA), HIF-1α and nitrite/nitrate were determined. In addition, Western blot analyses were performed to identify the amounts of Akt, endothelial nitric oxide synthase (eNOS) and ER stress parameters. RESULTS: IPC decreased cytolysis, lipid peroxidation and improved renal function. Parallelly, IPC enhanced Akt phosphorylation, eNOS, nitrite/nitrate and HIF-1α levels as compared to I/R group. Moreover, our results showed that IPC increased the relative amounts of glucose-regulated protein 78 (GRP78) and decreased those of RNA activated protein kinase (PKR)-like ER kinase (PERK), activating transcription factor 4 (ATF4) and TNF-receptor-associated factor 2 (TRAF2) as judged to I/R group. However, pre treatment with L-NAME abolished these beneficial effects of IPC against renal I/R insults. CONCLUSION: These findings suggest that early IPC protects kidney against renal I/R injury via reducing oxidative and ER stresses. These effects are associated with phosphorylation of Akt, eNOS activation and NO production contributing thus to HIF-1α stabilization. The beneficial impact of IPC was abolished when NO production is inhibited before IPC application.

Ischemic postconditioning does not attenuate ischemia-reperfusion injury of rabbit small intestine.

OBJECTIVE: To determine whether ischemic postconditioning can attenuate intestinal ischemia-reperfusion (I-R) injury and has a beneficial effect on tissue blood flow during reperfusion. STUDY DESIGN: In vivo experimental study. ANIMALS: New Zealand White rabbits (n=6). METHODS: Rabbits were anesthetized with pentobarbital, to avoid the preconditioning effects of volatile anesthetics, and ventilated with room air. Rectal temperature, hemodynamics, and normocapnia were maintained. After celiotomy, 3 jejunal segments were isolated in each rabbit for the following groups: (1) control, (2) I-R, and (3) I-R with postconditioning. I-R was induced by a 45-minute occlusion of the segment jejunal artery followed by 2-hour reperfusion. The postconditioning segment had 4 cycles of 30-second reperfusion and 30-second reocclusion during the initial 4 minutes of reperfusion. Stable isotope-labeled microspheres were used to measure intestinal blood flow at baseline, end occlusion, and end reperfusion. At the end of reperfusion, intestine segments were harvested and the rabbits euthanatized. A semiquantitative histopathologic evaluation (0-5) was conducted by a single, blinded observer. Wet-to-dry weight ratios were calculated to assess intestinal edema. RESULTS: There was no significant difference in grade of necrosis, tissue wet-to-dry weight ratios, or blood flow at any time point between ischemic and postconditioning groups. CONCLUSIONS: Ischemic postconditioning was ineffective in this model of intestinal I-R. CLINICAL RELEVANCE: Further experimental studies will need to be performed before clinical application of postconditioning for intestinal ischemia.