Shock/Sepsis/Trauma/Critical Care
Protective Effect of Rutin on the Ischemia/Reperfusion Induced Damage in Rat Kidney

https://doi.org/10.1016/j.jss.2009.03.022Get rights and content

Reactive oxygen species (ROS) are suggested to participate in ischemia/reperfusion (I/R) injury in the kidney. This study was designed to investigate the effect of rutin, a bioflavonoid, in I/R induced renal injury. Wistar albino rats were unilaterally nephrectomized, and 2 wk later they were subjected to 45min of left renal pedicle occlusion followed by 3h of reperfusion. Either rutin (1g/kg) or saline was administrated (i.p.) 1h prior to ischemia. At the end of the reperfusion period, kidney samples were taken for determination of renal malondialdehyde (MDA) and glutathione (GSH) levels, manganese-superoxide dismutase (MnSOD) activity and histological examination. Serum creatinine, blood urea nitrogen (BUN), and lactate dehydrogenase (LDH) concentrations were measured for the evaluation of renal function. I/R caused a significant decrease in GSH level and MnSOD activity, which was accompanied by a significant increase in MDA level of kidney tissues. Similarly, serum BUN and creatinine levels, as well as LDH were elevated in the I/R group compared with the control group. Pretreatment of rats with rutin (1g/kg/ i.p.) significantly attenuated renal dysfunction, reduced elevated MDA levels, and restored the depleted MnSOD activity and GSH levels. These beneficial changes in the biochemical parameters were also associated with parallel changes in histopathological appearance. These findings suggest that ROS play a causal role in I/R induced renal injury, and that rutin exerts renal-protective effects, probably by inhibiting ROS and antioxidant activities.

Introduction

Renal ischemia due to arterial occlusion or organ transplantation is a common cause of renal cell death and acute renal failure (ARF) and, in the case of transplantation, of delayed graft function or graft rejection [1]. Reperfusion aggravates the damage [2]. The mechanisms proposed to explain I/R injury include anoxia, release of reactive oxygen species (ROS) such as superoxide radical (O2), hydrogen peroxide (H2O2), and hydroxyl radical (–OH) during reperfusion, neutrophil accumulation, and subsequent release of additional ROS and lytic enzymes [3]. ROS react with biomolecules such as cell membrane lipid as well as proteins, carbohydrates, nucleic acids, and thiols resulting in organic radical formation, lipid peroxidation, enzyme inactivation, glutathione oxidation, and cell destruction 4, 5. Several studies have demonstrated that ROS have an important role in I/R injury, especially through lipid peroxidation 1, 6. They can lead to cellular injury by attacking membranes through peroxidation of polyunsaturated fatty acids, which can alter both membrane structure and function of mitochondrial, lysosomal, and plasma membranes [3]. Cellular defense against oxidative injury is provided by several mechanisms. Antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT), as well as nonenzymatic compounds such as reduced glutathione (GSH), ascorbic acid (AA), and α-tocopherol all help to cope with potential damage [7]. GSH is present in all mammalian cells, especially in renal cells, hepatocytes, and erythrocytes [8]. The increased production of ROS during I/R injury results in consumption and depletion of endogenous antioxidants. In this situation, the cells require exogenous antioxidant to protect them from ROS-induced damage.

Rutin (quercetin-3-rhamnosyl glucoside) is a kind of flavonoid glycoside found in buckwheat, many vegetables, fruits, and plant-derived beverages such as tea and wine [9]. Rutin is also known as Vitamin P and has antiplatelet, antiviral, and antihypertensive properties, as well as strengthing the capillaries, which is the result of its high radical scavenging activity and antioxidant capacity [10]. In addition, hypolipidaemic, cytoprotective antispasmotic and anticarcinogenic activities have also been reported. These properties are beneficial in preventing diseases and protecting the stability of the genetic material [11]. The micronucleus (MN) assay is an efficient biomarker for diagnosing genome instability in the cell [12]. Fenech [13] has suggested that MN frequency can be normalized or reduced on supplementation with specific micronutrients such as rutin, a-tocopherol, ascorbic acid. and that there is an optimal level of micronutrient intake for minimizing genome damage rate. Furthermore, La Casa et al. [14] clearly indicate that rutin significantly reduced the gastric mucosal damage produced by intragastric instillation of the necrotizing agent, and increased GPx activity. Robak and Gryglewski [15] have shown that rutin is a scavenger of the SOD-sensitive free radicals, which are generated during the activity of xanthine-oxidase. Dugas et al. [16] measured the antioxidant activity of a series of flavonoids against peroxyl radicals generated. In their study, the most active compound was the quercetin, followed by rutin. They suggest that potential role for dietary intake of rutin and quercetin containing foods in lowering the risk of certain pathophysiologies that have been associated with free radical-mediated disease. There are also studies that show a dose-response effect in inhibiting low-density lipoprotein (LDL) peroxidation of rutin 17, 18 Moreover, Milde et al. [19] suggested that rutin is a promising flavonoid for reducing the risk of atherosclerosis due to its inhibiting on LDL oxidation.

Despite all that, there is no study designed to investigate the effect of rutin on I/R damage in the rat kidney. Thus, the present study was undertaken to investigate the effect of rutin on I/R induced ARF in the kidney of male rats, by determining biochemical parameters and by histological examination.

Section snippets

Materials

Rutin, Na2EDTA, TCA, KCl, Trizma Base, DTNB, SDS, TBA, and 1,1,3,3 TEP were obtained from Sigma-Aldrich, Inc. (St. Louis, MI). Acetic acid, methanol, ethanol, xylene, entellane were obtained from Merck and Company (Whitehouse, NJ). Protein assay kit was purchased from BioRad Laboratories (Hercules, CA). All other chemicals were of analytical grade and purchased from local commercial sources.

Experimental Animals

In this study, 40 male Wistar albino rats weighing 250–300 g were used. The rats were obtained from the

Renal Function Studies

The creatinine, BUN, and LDH levels are shown in Table 1. Previous uninephrectomy did not alter the creatinine, BUN, and LDH levels in the Sham + NP group compared with the Sham group. Animals that underwent renal I/R exhibited significant increases (P < 0.05) in the serum concentration of creatinine compared with the Sham and Sham + NP groups, suggesting a significant degree of glomerular dysfunction. This was reflected by a significant increase (P < 0.05) in serum levels of BUN, also indicating

Discussion

The ARF produced by I/R is a clinical and experimental syndrome characterized by renal dysfunction, extensive tubular damage, tubular cell necrosis, and glomerular injury 25, 26. ROS are believed to play a central role in I/R injury in addition to several other factors such as neutrophils, platelets, reactive nitrogen species, the coagulation system, and xanthine-oxido-reductase enzyme system. ROS exert many deleterious effects on renal tissue [27], because they react with proteins, membrane

Acknowledgments

This work was supported by grants from Hacettepe University Research Foundation. This research is a part of the Ph.D. thesis of A. Korkmazc.

The authors thank Professor Dr. Şevket Ruacan for his support in histopathologic investigation. The authors also thank to Karen McDonough for her critical reading of the manuscript.

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