The protective effects of carnosine and melatonin in ischemia-reperfusion injury in the rat liver
Introduction
Ischemia starts a series of biochemical reactions that cause cell dysfunction and may progress to cell death. The restoration of blood flow to the ischemic tissue is requisite to remove toxic metabolites produced during the ischemic period. However, severe metabolic disorders may appear due to the infusion of toxic metabolites and various inflammatory mediators that are formed during ischemia into the systemic circulation and reperfusion may cause further tissue damage (Carden and Granger, 2000). Ischemia-reperfusion damage (IRD) is the cellular damage that appears due to the restoration of oxygen delivery to the hypoxic organ. IRD in the liver was first observed in the liver transplant that was experimentally performed by Toledo-Pereyra et al. (1975). Congestion, progressive thrombosis and graft necrosis leading to organ failure developed in the transplanted liver (Bilzer and Gerbes, 2000). The pathophysiology of liver IRD consists of many mechanisms that cause liver damage. Various cellular and molecular interactions such as Kupffer cell activation, formation of reactive oxygen species (ROS), cytokine and chemokine secretion, vasoconstriction, impairment of nitric oxide and endothelin balance, accumulation of neutrophil leukocytes, alteration in mitochondrial permeability, unbalanced influx of calcium (Ca2+) into the cell and pH paradox may be involved in the process. These complex mechanisms cause cell death, organ dysfunction and eventually organ loss (Kaplowitz, 2000; Jaeschke, 2003; Teoh and Farrell, 2003).
Carnosine (CAR) (β-alanyl-l-histidine), first described in the 1900s, was the first discovered of all neuropeptides (Gulewitsch and Amiradzibi, 1900). Muscles and nerves are diffusely distributed in tissues. Water soluble CAR functions in the cytosol where the oxidation mediators (metals and oxygen radicals) are abundantly located. Due to its biological function of scavenging active oxygen radicals, it has an antioxidant property. It acts as a scavenger of hydroxyl and superoxide radicals and, most effectively, of the singlet oxygen molecule. CAR has been observed to have a protective effect, based on its scavenging effect on hydroxyl and superoxide radicals, in brain, kidney, liver and skeletal muscle (Boldyrev et al., 1999; Fouad et al., 2007; Fujii et al., 2003, Fujii et al., 2005; Stvolinsky et al., 1999, Stvolinsky et al., 2000; Zalesova and Kuleva, 1998).
Pineal gland is the main source of melatonin (MEL) (N-acetyl-5-metoksitriptamin) in the circulation. It is also produced in small amounts in the retina, gastrointestinal system and by leukocytes (Reiter et al., 2007). MEL is tiny (Reiter et al., 2001a, Reiter et al., 2001b; Reiter and Tan, 2004) and highly lipophilic, and for this reason, it is found abundantly in all parts of the cell. MEL protects the DNA, lipids and proteins against oxidative damage (Reiter et al., 2000, Reiter et al., 2001a, Reiter et al., 2001b; Tan et al., 1993). The free radical scavenging and antioxidant effects of MEL have been shown in many studies (Cuzzocrea et al., 2000; Reiter and Maestroni, 1999; Cabeza et al., 2001; Pei et al., 2002).
In the present study, we aimed to compare the protective effects of CAR with MEL against liver IRD through histological (hematoxylin and eosin), immunohistochemical (apostain), electron microscopical and biochemical (ALT, alanine aminotransferase; AST, aspartate aminotransferase; GSH, glutathione; MPO, myeloperoxidase) parameters.
Section snippets
Animals and tissues
Female Wistar albino rats, weighing 200–250 g, were obtained from the Animal Research Laboratory of Dokuz Eylül University Medical School. The animals were synchronized to a 12:12 h light–dark cycle and housed at 20–22 °C before the beginning of the experiment. The animals were fed with a standard pellet diet and tap water ad libitum. The experiment protocol was approved by the Ethics Committee of Research on Laboratory Animals of Dokuz Eylül University Medical School.
Five study groups, each
Light microscopic examination
In the IR group, hepatocyte necrosis areas randomly disseminated in the liver parenchyma with disintegrated cell cordons, neuthrophil infiltration, expansion of sinusoids and congestion were detected. Hepatocyte damage was not observed in the liver parenchyma of the IR+CAR group except for isolated necrotic hepatocytes. Sinusoid expansion was much decreased compared to the IR group. It was observed that parenchymal integrity was maintained in the IR+MEL group. Expansion of sinusoids and cell
Discussion
Liver pathophysiology consists of many mechanisms that have an impact on liver damage at different levels. These mechanisms include cellular and molecular interactions, Kupffer cell activation, ROS formation, cytokine and chemokine secretion, vasoconstriction, nitric oxide, imbalance between endothelin and nitric oxide, accumulation of neutrophil leukocytes, alteration in the mitochondrial permeability, balanced calcium influx into the cell and pH paradox. These complex mechanisms result in
Acknowledgment
The authors acknowledge the financial support from Dokuz Eylul University Research Foundation Grant no. 04.KB.SAG.090.
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