Flurbiprofen, a Cyclooxygenase Inhibitor, Protects Mice from Hepatic Ischemia/Reperfusion Injury by Inhibiting GSK-3β Signaling and Mitochondrial Permeability Transition

Abstract

Flurbiprofen acts as a nonselective inhibitor for cyclooxygenases (COX-1 and COX-2), but its impact on hepatic ischemia/ reperfusion (I/R) injury remains unclear. Mice were randomized into sham, I/R and flurbiprofen (Flurb) groups. The hepatic artery and portal vein to the left and median liver lobes were occluded for 90 min and unclamped for reperfusion to establish a model of segmental (70%) warm hepatic ischemia. Pretreatment of animals with flurbiprofen prior to I/R insult significantly decreased serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH), and prevented hepatocytes from I/R-induced apoptosis/necrosis. Moreover, flurbiprofen dramatically inhibited mitochondrial permeability transition (MPT) pore opening, and thus prevented mitochondrial-related cell death and apoptosis. Mechanistic studies revealed that flurbiprofen markedly inhibited glycogen synthase kinase (GSK)-3β activity and increased phosphorylation of GSK-3β at Ser9, which, consequently, could modulate the adenine nucleotide translocase (ANT)-cyclophilin D (CyP-D) complex and the susceptibility to MPT induction. Therefore, administration of flurbiprofen prior to hepatic I/R ameliorates mitochondrial and hepatocellular damage through inhibition of MPT and inactivation of GSK-3β, and provides experimental evidence for clinical use of flurbiprofen to protect liver function in surgical settings in addition to its conventional use for pain relief.

Hydrogen Sulfide Preconditioning Protects Rat Liver against Ischemia/Reperfusion Injury by Activating Akt-GSK-3β Signaling and Inhibiting Mitochondrial Permeability Transition

“…However, there is no direct evidence that H 2 S has an effect on systemic dynamics. Our study confirmed that intravenous injection of 25 μmol/kg NaHS had no effect on systemic hemodynamics at various time points in a rat model of 70% warm hepatic I/R, which is widely used in studies focused on hepatic I/R [25,28,44,45,46,47]. Given that the hepatic portal system was not completely blocked (with the blood supply maintained in the right lobe and the caudate lobe), the blood returns from the postcava to the right atrium unaffected.…”

“…Mitochondria were isolated by gradient centrifugation as we previously described [25]. Briefly, fresh liver tissues (1 g) were homogenized with 8 ml of isolation buffer containing 220 mmol/L D-mannitol, 70 mmol/L sucrose, 10 mmol/L Tris-HCl, 1 mmol/L EGTA, and 0.4% bovine serum albumin (pH 7.4).…”

“…A calcium retention capacity (CRC) assay was adapted from a previously described method [25]. Briefly, the CRC was defined as the amount of Ca 2+ required to trigger a massive Ca 2+ release in isolated liver mitochondria.…”

“…Therefore, sodium hydrosulfide (NaHS) can be administered as a water-soluble H 2 S donor. The diverse physiological functions of H 2 S make it capable of protecting the heart [19], brain [20], liver [21,22,23], kidney [24], and lung [25] against I/R injury when given at sub-toxic doses. In the liver, the underlying mechanisms of protection appear to include suppressing oxidative stress via antioxidant activities, reducing inflammatory mediators, such as tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10) and intercellular cell adhesion molecule-1 (ICAM-1), and reducing hepatocyte apoptosis.…”

Silencing of Long Noncoding RNA AK139328 Attenuates Ischemia/Reperfusion Injury in Mouse Livers

“…As one main downstream molecule of Akt, GSK-3 plays an important role in liver ischemia/reperfusion injury. Inhibition of GSK-3 has been shown to ameliorate liver ischemia/reperfusion injury [40-42]. In this study, we found that silencing of AK139328 resulted in inhibition of GSK-3 in ischemia/reperfusion treated mouse livers, which is consistent with Akt activation.…”