Reearch Article

Autin Therapeutic. 2014;1(2): 7.

Selenium Supplementation to Chronic Kidney Dieae Patient on Hemodialyi ha no Effect on Superoxide Dimutae Activity and Malonyldialdehyde Concentration in Blood

Zachara BA1,2*, Gromadzinka J1, Waowicz W1, Swiech R3 and Zbrog Z3

1Departament of Toxicology and Carcinogenei, Nofer Intitute of Occupational Medicine, Poland

2Higher School of Health Science, Bydgozcz, Poland

3B Braun Avitum Dialyi Center, Lodz, Poland

*Correponding author: BA Zachara, Departament of Toxicology and Carcinogenei, Nofer Intitute of Occupational Medicine, Lodz, 33/67 Nowodworka St, 85120 Bydgozcz, Poland

Received: September 20, 2014; Accepted: November 07, 2014; Publihed: November 11, 2014

Abtract

Background: Hemodialyi (HD) i the mot common form of treating patient in the End-Stage Renal Dieae (ESRD). The mot common approach to meauring the effect of oxidative tre ha been to meaure the activity of Glutathione Peroxidae (GSH-Px), Superoxide Dimutae (SOD) and the product of lipid peroxidation &ndah; Malonyldialdehyde (MDA). In Chronic Kidney Dieae (CKD) patient&rquo; Selenium (Se) level in blood i frequently lower than in healthy ubject and it decreae gradually with the progre of the dieae.

Aim: The aim of our tudy wa to examine whether the adminitration of Se to patient on HD alter the activity of GSH-Px and SOD in red blood cell and the level of MDA in plama.

Patient and Method: Our tudy involved 3 group of ubject: 1) 52 CKD nondialyzed patient, 2) patient in ESRD upplemented for 3 month with Se-rich yeat, 200 μg/day (n = 30) or placebo (n = 28) and 3) 52 healthy ubject. The GSH-Px and SOD activitie in red blood cell hemolyate and lipid peroxidation product [expreed a Thiobarbituric Acid Reactive Subtance (TBARS)] in plama were aayed.

Reult: GSH-Px activity in RBC wa ignificantly lower in nondialyzed CKD patient a compared with control group, but wa even more reduced in patient on HD. SOD activity in red blood cell wa ignificantly lower in patient in ESRD than in the healthy ubject and in the nondialyzed patient. Se upplementation to the HD patient ha no effect on the change in SOD activity. TBARS level in plama in patient on HD wa ignificantly higher than in the control group and in nondialyzed patient. Se adminitration did not reduce thi level.

Concluion: Se upplementation to patient on HD ha no effect on the activity of SOD in red blood cell and doe not prevent lipid peroxidation.

Keyword: Chronic kidney dieae; Hemodialyi; Selenium upplementation; Superoxide dimutae; Malonyldialdehyde

Introduction

Oxidative tre i defined a an imbalance between prooxidant and antioxidant in favor of the oxidant, potentially leading to damaging biological ytem [1,2]. Oxidative tre i preent and it marker can be meaured in both healthy people and thoe with variou clinical diorder [3]. Oxidative tre ha been linked with damaged protein, DNA, lipid in cell membrane (mainly unaturated fatty acid; PUFA) and carbohydrate [3], and thu lead to the progreion of everal dieae including cardiovacular dieae, cancer, Chronic Kidney Dieae (CKD) and other [4]. Mot importantly, oxidative tre i believed to promote the endothelial dyfunction and atherocleroi and, therefore, cardiovacular complication [5]. Free radical formed during oxidative tre are alo reponible for DNA damage and, a a conequence, for cancer development [6-8]. Long period of Hemodialyi (HD) treatment are linked to DNA damage due to oxidative tre [9]. The relationhip between DNA damage and cancer development ha been widely documented [10].

Hemodialyi i the mot common form of treatment for End-Stage Renal Dieae (ESRD) patient, and i aociated with coniderable mortality due to cardiovacular dieae and cancer [11,12]. The mot common approach to the meaurement of oxidative tre and free radical ha been to meaure the product of lipid peroxidation and PUFA oxidation &ndah; the level of Malonyldialdehyde (MDA) [13], a well a the activity of antioxidant enzyme. The body&rquo; defene againt lipid peroxidation include the enzyme: uperoxide dimutae (SOD; EC 1.15.1.1), glutathione peroxidae (GSH-Px; EC 1.11.1.9) and catalae (CAT; EC 1.11.1.6) [3]. Thee enzyme detroy dangerou product of oxygen metabolim.

Superoxide dimutae play a central role in catalyzing the pontaneou dimutation of uperoxide (O2·-) into oxygen and hydrogen peroxide [14]. Thu, they are an important antioxidant defene in nearly all cell expoed to oxygen. Hydrogen peroxide i then detroyed by GSH-Px and CAT [15].

Selenium (Se) i an eential trace element required in microgram amount by all mammal. It i incorporated in the form of Selenocyteine (Sec) into 25 elenoprotein [16]. Enzymatic activity ha been aigned to 12 of thee elenoprotein, and ome of them have antioxidant activitie [17]. GSH-Px i the mot extenively characterized elenoprotein, being found in Red Blood Cell (RBC) and cytool of nearly all tiue of mammal, bird and everal other organim. It i called claical GSH-Px (cGSH-Px or GSH-Px1) [18-20]. Five ioform of GSH-Px have been identified [21], two are preent in the blood: GSH-Px 1 preent in red blood cell and GSHPx 3 preent in plama. Both have a tetrameric form and contain one elenium per ubunit (or four gram atom of Se per mole of enzyme) in the form of Sec [22-24].

Zinc (Zn) i incorporated in the catalytic ite of everal hundred metaloprotein [25]. Copper (Cu) i an eential element for all living organim, erving a a cofactor for many important metaloprotein and enzyme [26]. Cu and Zn form the active ite of one of the form of SOD &ndah; Cu2+/Zn2+ SOD (called SOD 1), preent in cytool and red blood cell [27]. Concentration of Se and Zn in blood plama of CKD patient are decreaed [28,29] but the tatu of copper doe not eem to be influenced by CKD [30].

The aim of thi tudy wa to determine the activity of SOD and GSH-Px in RBC, the level of Zn, Cu and MDA concentration in plama of HD patient upplemented for 3 month with 200 μg of Se per day.

Material and Method

Patient and control

A 3-month, randomized double-blind, placebo-controlled trial wa carried out. The tudy involved 3 group of ubject. Group 1 compried of 52 of CKD nondialyzed patient in different tage of the dieae (creatinine level: 1. 00 &ndah; 10.99 mg/dL; mean: 5.16 mg/ dL); group 2: 58 CKD patient in ESRD (creatinine level: 4.20 &ndah; 16.60 mg/dL; mean: 9.43 mg/dL) on regular HD, divided into 2 ubgroup: 30 patient upplemented with 200 μg Se/day in the form a high- Se yeat tablet (produced by Pharma Nord, Bioelenium, Denmark) for 3 month, and 28 patient upplemented with placebo tablet containing identical yeat with no added Se (Pharma Nord). The patient were dialyzed 3 time a week for 4 hour. Group 3 conited of 52 healthy ubject. The tudy wa approved by the Intitute Ethic Committee for Medical Reearch (No. 18/2003) and all the participant gave their written conent.

Method

Blood ample were drawn from all the participant into vacutainer tube containing lithium heparin a an anticoagulant. From the healthy control and the patient with CKD not on dialyi, blood wa taken once, and from the patient undergoing dialyi, three time: before tarting the tudy and after 1 and 3 month of tablet upplementation. Blood wa centrifuged (+4o C, 5 000 r.p.m., 10 min), the plama wa harveted and tored at &ndah;20oC until analyi. The red blood cell were wahed three time with an exce of chilled 0.9% aline olution and were then hemolyzed by freezing and thawing and centrifuged again. Hemoglobin wa meaured by the routine cyanmethemoglobin method. Creatinine concentration wa determined by routine laboratory method uing Jaffy reaction (a kit produced by Cormay, Lublin, Poland). SOD activity in RBC hemolyate wa determined according to the method of Beauchamp and Fridovich [31] and wa expreed in U/g Hb. The GSH-Px activity in red cell hemolyate wa aayed by the coupled method of Paglia and Valentine [32] uing tert-butyl hydroperoxide a a ubtrate. One unit of the enzyme activity wa expreed a 1 mol NADPH oxidized/ min/g Hb of hemolyate (U/g Hb). Lipid peroxidation in the plama wa monitored by determining the end product of lipid peroxidation &ndah; malonyldialdehyde &ndah; decribed by Waowicz et al [33]. The value were expreed a Thiobarbituric Acid Reactive Subtance (TBARS)in nmol/mL. The concentration of Zn and Cu were meaured by flame atomic aborption pectrometry [34] uing Pye Unicam SP9 800 apparatu. The accuracy of the method wa checked with erum reference material (Seronorm, Nycomed, lot 704121).

Statitical analyi

Comparion of the level under tudy were made by multivariate analyi of variance [35] at three time point (before the tudy, one month and three month after tudy). When ignificant difference were found between the group, the difference were teted at all time point. The tet were baed on Shapiro-Wilk&rquo; tatitic, ignificance being et at 0.05. All tatitic were conducted uing the STATA 9 package.

Reult

GSH-Px activity in RBC wa ignificantly lower in the patient with CKD not on dialyi (P < 0.02) a compared with the control, but wa even more reduced in the patient on HD (P < 0.0001) (Figure 1). Dialyi treatment led to an increae in GSH-Px activity in both ubgroup, however the increae in activity wa much higher (P < 0.0001) in the ubgroup upplemented with Se in comparion with the placebo group (P < 0.005).

Figure 1: GSH-Px activity in RBC of the healthy control, the CKD nondialyzed patient and the patient on hemodialyi upplemented with placebo (white column) and elenium (dark column). Statitic: a, the CKD patient v. control, P < 0.02; b, the HD 0 (both ubgroup taken together) v. the control and the nondialyzed CKD patient, P < 0.0001; c, HD 3 v. HD 0 (upplemented with placebo), P < 0.005; d, HD 3 v. HD 0 (upplemented with Se), P < 0.0001.

    
    
Figure 1: GSH-Px activity in RBC of the healthy control, the CKD nondialyzed patient and the patient on hemodialyi upplemented with placebo (white column) and elenium (dark column). Statitic: a, the CKD patient v. control, P < 0.02; b, the HD 0 (both ubgroup taken together) v. the control and the nondialyzed CKD patient, P < 0.0001; c, HD 3 v. HD 0 (upplemented with placebo), P < 0.005; d, HD 3 v. HD 0 (upplemented with Se), P < 0.0001.
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SOD activity in RBC of the nondialyzed patient with CKD did not differ from the activity in the healthy control. In the dialyzed patient, at the baeline (both ubgroup taken together), SOD activity wa lower only by 8.5% compared with control (P < 0.02) (Figure 2). Hemodialyi lead to a mall reduction in enzyme activity in both ubgroup, but after 3 month, thee value were not ignificantly different from the activity found at HD 0. It hould be emphaized that elenium upplementation to the patient on HD had no effect on the change in the activity of thi enzyme.

Figure 2: SOD activity in RBC of the healthy control, the nondialyzed CKD patient and the patient on hemodialyi upplemented with placebo (white column) and elenium (dark column). Statitic: a, the HD 0 (both ubgroup) v. the control and the nondialyzed CKD patient, P < 0.02; b, HD 3 v. HD 0, P = 0.1 (NS).

    
    
Figure 2: SOD activity in RBC of the healthy control, the nondialyzed CKD patient and the patient on hemodialyi upplemented with placebo (white column) and elenium (dark column). Statitic: a, the HD 0 (both ubgroup) v. the control and the nondialyzed CKD patient, P < 0.02; b, HD 3 v. HD 0, P = 0.1 (NS).
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Zinc concentration in plama of the nondialyzed CKD patient and the patient on HD wa ignificantly lower compared with the healthy control (P < 0.0001) (Figure 3). In the dialyzed patient the concentration wa lower than in the nondialyzed (P < 0.05). In the placebo group it did not change during the 3 month tudy. In the dialyzed patient Se upplementation led to a gradual increae in the concentration of Zn, which after 3 month had reached a valueignificantly higher compared with the HD 0 (P < 0.01).

Figure 3: Zinc concentration in plama of the healthy control, the nondialyzed CKD patient and the patient on hemodialyi upplemented with placebo (white column) and elenium (dark column). Statitic: a, the CKD nondialyzed patient and the HD 0 (both ubgroup taken together) v. the control, P < 0.0001; b, HD 0 (both ubgroup) v. CKD, P < 0.05; c, HD 3 (+Se) v. HD 0 (+Se), P < 0.01.

    
    
Figure 3: Zinc concentration in plama of the healthy control, the nondialyzed CKD patient and the patient on hemodialyi upplemented with placebo (white column) and elenium (dark column). Statitic: a, the CKD nondialyzed patient and the HD 0 (both ubgroup taken together) v. the control, P < 0.0001; b, HD 0 (both ubgroup) v. CKD, P < 0.05; c, HD 3 (+Se) v. HD 0 (+Se), P < 0.01.
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Copper concentration in plasma in all groups was nearly the same and ranged from 1.27 to 1.37 mg/L (Figure 4). The values in both subgroups remained constant during the course of the dialysis.

Citation: Zachara BA, Gromadzinska J, Wasowicz W, Swiech R and Zbrog Z. Selenium Supplementationto Chronic Kidney Disease Patients on Hemodialysis has no Effect on Superoxide Dismutase Activity and Malonyldialdehyde Concentration in Blood. Austin Therapeutics. 2014;1(2): 7. ISSN:2472-3673