Selenium and high dose vitamin E administration protects cisplatin-induced oxidative damage to...

Toxicology 195 (2004) 221–230

Selenium and high dose vitamin E administration protectscisplatin-induced oxidative damage to renal,

liver and lens tissues in rats

Mustafa Nazıroglua,∗, Aziz Karaoglub, Asude Orhan Aksoyb

a Department of Physiology, Veterinary Faculty, Fırat University, TR-23119, Elazıg, Turkeyb Department of Oncology, Medical Faculty, Fırat University, TR-23119, Elazıg, Turkey

Received 19 August 2003; received in revised form 23 October 2003; accepted 23 October 2003

Abstract

Cisplatin is one of the most active cytotoxic agents in the treatment of cancer but its clinical use is associated with nephrotoxicity.Several studies suggest that supplementation with antioxidant can influence cisplatin induced nephrotoxicity. In the present study,we investigated the effect of selenium with high dose vitamin E administration on lipid peroxidation (MDA) and scavengingenzyme activity in kidneys, liver and lens of cisplatin-induced toxicity in rats.

Forty female Wistar rats were used. They were randomly divided into five groups. The first and second groups were used ascontrol and cisplatin (6 mg/kg BW) intraperitoneally administrated groups. Groups III, IV and V received intraperitoneally fivedoses of selenium (1.5 mg/kg BW) and a high dose of vitamin E (1000 mg/kg BW) combination before, simultaneously andafter with cisplatin, respectively.

Glutathione peroxidase (GSH-Px), vitamin E and�-carotene levels in the kidney, lens and liver, vitamin A and reducedglutathione (GSH) levels in the kidney were significantly (P <0.05 to<0.001) lower in the cisplatin group than in the controlwhereas there was a significant increase in kidney, liver and lens MDA levels in rats treated with cisplatin. The decreasedantioxidant enzymes and vitamins and increased MDA levels in the kidney, lens and liver of animals administered with cisplatinwere significantly (P <0.05 to<0.001) improved with selenium and a high dose vitamin E injection.

In conclusion, this data demonstrates that there is an increase in lipid peroxidation in the kidney, liver and lens of animalsadministered with cisplatin whereas there is a decrease in antioxidant vitamins and enzymes. However, intraperitoneally injectedselenium combined with a high dose of vitamin E seem to produce a significant improvement on antioxidants concentrations inrats treated before, simultaneously and after with cisplatin. The selenium with high dose vitamin E injection may play a role inpreventing cisplatin-induced nephropathy and cataract formation in cancer patient.© 2003 Elsevier Ireland Ltd. All rights reserved.

Keywords: Oxidative stress; Cisplatin-nephrotoxicity; Lens; Liver; Selenium; Vitamin E

Abbreviations: GSH, reduced glutathione; GSH, Px, glutathione peroxidase; MDA, malondialdehyde; RBC, red blood cells; ROS,reactive oxygen species; SOD, superoxide dismutase; SeVE, selenium and high dose vitamin E

∗ Corresponding author. Present address: Klinikum der RWTH, Institut für Physiologie, Pauwelsstr. 30, 52057 Aachen, Germany.Tel.: +49-241-8088-812; fax:+49-241-8082-434.

E-mail address: [email protected] (M. Nazıroglu).

0300-483X/$ – see front matter © 2003 Elsevier Ireland Ltd. All rights reserved.doi:10.1016/j.tox.2003.10.012

222 M. Nazıroglu et al. / Toxicology 195 (2004) 221–230

1. Introduction

Cisplatin (Cis-diamminedichloroplatinum II) showspromise as an antitumor agent for several types ofcancer and has been widely used for chemotherapy.Activity has been demonstrated against a varietyof tumors, particularly for head and neck, testic-ular, ovarian, bladder and small cell lung cancers(Weijl et al., 1997; Kuhlmann et al., 1997). However,high-doses administered to patients produce nephro-toxic and hepatotoxicy side effects, and the dose ofcisplatin must often be limited (Yoshida et al., 2000).The alterations in the kidney and liver functionsinduced by cisplatin are characterized by signs of in-jury, such as glutathione status, and cisplatin-inducednephropathy is closely associated with an increasein lipid peroxidation (Weijl et al., 1997; Antunesand Darin, 2000; Antunes et al., 2001; Mora et al.,2003).

The pathogenesis of renal damage caused by cis-platin is generally considered to be oxidative damage(Yoshida et al., 2000). Administration of cisplatincauses an increase in lipid peroxide (MDA) levels anda decrease in the activity of enzymes that prevent orprotect against lipid peroxidation in the tissues. There-fore, the administration of antioxidants such as ebselen(Yoshida et al., 2000), vitamin C (Antunes and Darin,2000) and selenium (Caffrey and Frenkel, 2000;Antunes et al., 2001) before treatment with cisplatinhas been used to protect against nephrotoxicity inhuman and experimental animals. In the kidney, thesetreatments are reported to diminish the increase inMDA and the decrease in protective enzyme activitythat are induced by cisplatin.

The selenium is an essential dietary trace elementwhich plays an important role in a number of bio-logical process (Combs and Combs, 1986). There isa long-standing association between selenium com-pounds and cancer chemo-prevention (Conklin, 2000).Many experimental studies in animals have demon-strated the ability of selenium to prevent carcinogen-esis, and epidemiological studies have suggested thata decreased selenium status in humans is associatedwith an increased risk of cancer (Weijl et al., 1997). Arecent widely publicized chemo-prevention study hasshown that selenium supplements can decrease the in-cidence of certain types of cancer (Conklin, 2000). Inaddition, selenium in one study selenium (Caffrey and

Frenkel, 2000) was utilized in preventing the nephro-toxic side effects of cisplatin although selenium in an-other selenium study (Antunes et al., 2001) had noeffect.

The most frequently studied antioxidant vitaminsare vitamins A and E. Vitamin A serves a prohormonefor retroretinoids and intersects with signal transduc-tion at cytoplasmic and membrane sites (Czernichowand Hercberg, 2001). Vitamin E is the major lipophilicchain-breaking antioxidant present within cell mem-branes (Packer and Landvik, 1990). In contrast to vi-tamins A and C, vitamin E can be safely used inhigh doses in the prevention of diseases such as di-abetes and cardiovascular disease (Czernichow andHercberg, 2001). It suppresses the oxidative stress ofmembranes. Given to potential role of the ROS in me-diating tissue damage, cells contain The enzymes, su-peroxide dismutase (SOD) and glutathione peroxidase(GSH-Px), in the cells are important components ofseveral naturally occurring antioxidant defense mech-anisms to prevent oxidative injury (Weijl et al., 1997).There is also a hypothesis that decreased levels of nat-ural antioxidants vitamins and diminished scavengingenzyme capacity may be responsible for the excessof ROS observed in cisplatin-induced toxicity (Weijlet al., 1997; Antunes and Darin, 2000; Antunes et al.,2001). Supplementation of the antioxidant vitamin Cand selenium, has been reported to inhibit lipid per-oxide in various conditions such as cisplatin-inducednephrotoxicity (Caffrey and Frenkel, 2000; Antunesand Darin, 2000).

If cisplatin increase MDA, increased amounts ofenzymatic and non-enzymatic antioxidants should beoxidized and their levels in kidney, lens and livershould be diminished. To test this hypothesis, ratswere studied after cisplatin administration to ascertainwhether cisplatin in rats increases MDA and rate ofkidney, liver and lens enzymatic and non-enzymaticantioxidants disappearance. The second aim of thecurrent study was to test the effects of a moderatedose selenium and high dose vitamin E (SeVE) andits possible beneficial effect on antioxidant defensesystem by evaluating MDA and scavenging enzymeactivity in rats administrated before or simultaneouslyor after with cisplatin. A combination of vitamin Ewith selenium was chosen since vitamin E, is a freeradical scavenger (Weijl et al., 1997), and it wasfound,Antunes et al. (2001)informed that a selenium

M. Nazıroglu et al. / Toxicology 195 (2004) 221–230 223

supplementation on its own did not have protectiveeffects against cisplatin-induced toxicity.

2. Materials and methods

2.1. Chemicals

All chemicals were obtained from Sigma Chemi-cal Inc. (St. Louis, MO) and all organic solvents andsodium selenite from Merck Chemical Inc. (Darm-standt, Germany). The inject able form of vitamin E(dl-�-tocopheryl acetate) was obtained from F. Hoff-man La Roche (Istanbul, Turkey). The inject able formof cisplatin was purchased from Eczacibasi (Istan-bul, Turkey). All reagents were of analytical grade.All reagents except the phosphate buffers were pre-pared each day and stored in a refrigerator at+4◦C.The reagents were equilibrated at room temperaturefor 0.5 h before use when the analysis was initiatedor reagents containers were refilled. Phosphate bufferswere stabilised at+4◦C for 1 month.

2.2. Animals

Medical Faculty Experimentation Ethics Committeeof our university approved experimental proceduresof the study. Forty adult female Wistar rats bred inour laboratory were used. Housing was at 22–24◦Cwith light from 8:00 to 20:00 h with free access towater. At the start of the experiment, the rats aged12 weeks weighed 150–165 g. They were randomlydistributed into five groups and housed individuallyin stainless- steel cages in a pathogen-free UniversityLaboratory Animal Research facility. All animals werefed a commercial diet (Elazıg Feed Factory, Elazıg,Turkey) including the ingredients shown inTable 1during the experiment.

2.3. Study groups and sampling

Control I (n = 8): normal controls; received fivedoses of physiologic saline (2.5 ml per animal)intraperitoneally (i.p.).

Group A (n = 8): received cisplatin (5 mg/kg BW)and four doses of physiologic saline i.p. and theanimals were killed 5 days after a dose cisplatininjection.

Table 1Diet composition

Ingredients (%)

Corn 25.1Barley 20.0Soybean 36.0Wheat 9.7Fish flour 3.2Meat-bone flour 2.4Calcium-phosphate 1.7Salt 1.1Methionine 0.2Lysin 0.2Vitamin/mineral mixa 0.2

a In per kg mixture: vitamin A 12,000,000 IU; vitamin C 50 g;vitamin D3 2,400,000 IU; vitamin E 30 g; vitamin K3 2.5 g andvitamins B1 3 g; B2 7 g; B6 4 g and B12 15 mg; nicotinamide4 g; calcium-D pantothane 8 g; Folic acid 1 g; biotin 45 mg; Mn80 g; Fe 40 g; Zn 60 g; Cu 5 g; I 0.4 g; Co 0.1 g; Se 0.15 g andantioxidant (Butyhydroxytoluol) 10 g.

Group B (n = 8): received five doses of sele-nium (1.5 mg/kg BW) and high dose of vitaminE (1000 mg/kg BW) before with cisplatin and theanimals were killed 5 days after cisplatin injec-tion.

Group C (n = 8): received five doses of sele-nium (1.5 mg/kg BW) and high dose of vitaminE (1000 mg/kg BW) simultaneously with a dosecisplatin and the animals were killed 5 days aftercisplatin injection.

Group D (n = 8): received five doses of sele-nium (1.5 mg/kg BW) and high dose of vitaminE (1000 mg/kg BW) after with a dose cisplatinand the animals were killed 5 days after cisplatininjection.

The rats were anaesthetised with ether for 5 min.The kidney, liver and lens samples were removed, pro-tected against light. Liver, lens and kidney sampleswere washed three times in cold isotonic saline (0.9%(v/w)), then right kidney and lens samples were storedat −30◦C for <3 months pending measurement ofenzymatic activity. The left kidney and lens sampleswere used for immediate lipid peroxidation and vita-min assay.

2.4. Homogenate preparation

For the enzymatic analyses in lens (50 mg), kidneyand liver (250 mg) tissues were minced on a glass


and homogenized by a glass–glass homogenizator(Çalıskan glass technique, Ankara, Turkey) in coldphysiological saline on ice. Then the tissues werediluted with a nine-fold volume of phosphate buffer(pH 7.4).

2.5. Lipid peroxidation assay

Lipid peroxidation (as malondialdehyde, MDA)levels in kidney and lens homogenate were measuredwith the thiobarbituric-acid reaction by the methodof Placer et al. (1966)as described in previous stud-ies (Nazıroglu and Çay, 2001; Nazıroglu, 2003). Thequantification of thiobarbituric acid reactive sub-stances was determined by comparing the absorptionto the standard curve of malondialdehyde (MDA)equivalents generated by acid catalyzed hydrolysisof 1,1,3,3 tetramethoxypropane. The values of MDAwere expressed as�mol/g protein. Every sample wasassayed in duplicate, and the assay coefficients ofvariation for MDA were less than 3%.

2.6. Reduced glutathione (GSH), glutathioneperoxidase (GSH-Px) and protein assay

The GSH content of the kidney and lens ho-mogenate was measured at 412 nm using the methodof Sedlak and Lindsay (1968)as described own stud-ies (Nazıroglu and Çay, 2001; Nazıroglu, 2003). Thesamples were precipitated with 50% trichloraceticacid and then centrifuged at 1000× g for 5 min.The reaction mixture contained 0.5 ml of supernatant,2.0 ml of Tris–EDTA buffer (0.2 M; pH 8.9) and0.1 ml of 0.01 M 5,5′-dithio-bis-2-nitrobenzoic acid.The solution was kept at room temperature for 5 min,and then read at 412 nm on the spectrophotometer.GSH-Px activities of the kidney and lens were spec-trophotometrically measured at 37◦C and 412 nmaccording to theLawrence and Burk (1976).

The protein content in the kidney and lens was mea-sured by method ofLowry et al. (1951)with bovineserum albumin as the standard.

2.7. Plasma vitamins A and E and β-caroteneanalyses

Vitamins A (retinol) and E (�-tocopherol) were de-termined in the kidney samples by a modification of

the method described by method ofDesai (1984). Be-fore vitamin analyses liver, lens and kidney sampleswere minced on glass plate. Two hundred microgramof kidney homogenate samples was saponified by theaddition of 0.3 ml 60% (w/v in water) KOH and 2 mlof 1% (w/v in ethanol) ascorbic acid, followed byheating at 70◦C for 30 min. After cooling the sam-ples on the ice, 2 ml of water and 1 ml of n-hexanewere added and mixed with the samples and they thenrested for 10 min to allow phase separation. N-hexaneextract was dried under nitrogen at 40◦C and redis-solved using 0.2 ml methanol. Twenty-microliter por-tions of the methanol extracts were chromatographedon high-performance liquid chromatography. Fluo-rimetric detection of vitamin A used excitation andemission wavelengths of 330 and 480 nm, respec-tively. The relevant wavelengths for�-tocopheroldetection were 292 and 330 nm. Calibration was per-formed using standard solutions of all-trans retinoland�-tocopherol in methanol.

The levels of�-carotene in kidney samples weredetermined according to method ofSuzuki and Katoh(1990). Two milliliter of hexane mixed with 0.25 g tis-sue and absorbance of hexane was measured at 453 nmin the spectrophotometer.

2.8. Statistical analyses

All results are expressed as means± S.D. To de-termine the effect of treatment, data were analyzedusing one-way ANOVA repeated measures.P-valuesof less than 0.05 were regarded as significant. Signifi-cant values were assessed with Tukey’s multiple rangtest. Data was analyzed using SPSS statistical program(version 10.0 software, SPSS Inc. Chicago, Illinois,USA).

3. Results

3.1. Changes in antioxidant enzyme and MDA levelsand kidney weight

The changes in the MDA and GSH levels andGSH-Px activity in the kidney and lens were shownin Figs. 1–3. The MDA concentrations in the tissueswere used as a measure of lipid peroxidation. Whencompared to control groups MDA levels in the kidney,


Fig. 1. The effects of selenium and high dose vitamin E administration on MDA levels in the kidney, liver and lens of rats administeredcisplatin (A) and before (B), simultaneously (C) and after (D) with cisplatin. (mean± S.D., n = 8). (a) P < 0.05, (b) P < 0.01, (c)P < 0.001 as compared with group control. (d) P < 0.05, (e) P < 0.01, (f ) P < 0.001 as compared with group A.

liver, and lens were significantly (P < 0.01) higherin groups administered with cisplatin. On the otherhand, 5 days of SeVE administration was associatedwith a fall in the MDA levels of liver, lens and kidney.The effects of after treatment (D group) of femalerats with SeVE on cisplatin-mediated changes in thelevels of liver and kidney MDA was higher than inthe B and C groups.

Cisplatin toxicity changed with GSH-Px and GSHlevels in the kidney, liver and lens. GSH level in the

Fig. 2. The effects of selenium and high dose vitamin E administration on glutathione peroxidase activities in the kidney, liver and lensof rats administered cisplatin (A) and before (B), simultaneously (C) and after (D) with cisplatin. (mean± S.D., n = 8). (a) P < 0.05, (b)P < 0.01, (c) P < 0.001 as compared with group control. (d) P < 0.05, (f ) P < 0.001 as compared with group A.

kidney was significantly (P < 0.05) lower in the Agroup than in the control group although GSH-Px ac-tivity in the liver was not affected by cisplatin admin-istration. GSH-Px activities in the kidney, liver andlens and GSH levels in kidney were positively af-fected by SeVE supplementation. GSH-Px activity inthe kidney, liver and lens, GSH levels in the kidneywere significantly (P < 0.05 andP < 0.001) higherin the B, C and D groups than in both the control andA groups.


Fig. 3. The effects of selenium and high dose vitamin E administration on reduced glutathione levels in the kidney, liver and lens of ratsadministered cisplatin (A) and before (B), simultaneously (C) and after (D) with cisplatin. (mean± S.D., n = 8). aP < 0.05 as comparedwith group control.dP < 0.05 as compared with group A.

3.2. Changes in vitamin concentrations

The changes in vitamins A, E and�-carotene in thekidney are shown inFigs. 4–6. In this study, compar-ison was made between five groups (control, A, B, Cand D) for the parameter of�-carotene and vitamins Aand E concentrations. There were significant changesin the kidney vitamin E (P < 0.01) concentrations of

Fig. 4. The effects of selenium and high dose vitamin E administration on vitamin E concentrations in the kidney, liver and lens ofrats administered cisplatin (A) and before (B), simultaneously (C) and after (D) with cisplatin. (mean± S.D., n = 8). (a) P < 0.05, (b)P < 0.01, (c) P < 0.001 as compared with group control. (f ) P < 0.001 as compared with group A.

A, B, C and D groups. Vitamin E concentrations inliver, kidney and lens were significantly (P < 0.05 andP < 0.01) lower in the A groups and than in the con-trol group. However, vitamin E concentration in thekidney, liver and lens was significantly (P < 0.001)higher in the B, C and D groups than both in boththe control and A groups. Therefore, we observed thatcisplatin administration induced a vitamin E decreas-


Fig. 5. The effects of selenium and high dose vitamin E administration on vitamin A concentrations in the kidney and liver of ratsadministered cisplatin (A) and before (B), simultaneously (C) and after (D) with cisplatin. (mean±S.D., n = 8). (a) P < 0.05 as comparedwith group control. (d) P < 0.05 as compared with group A.

Fig. 6. The effects of selenium and high dose vitamin E administration on�-carotene concentration in the kidney, liver and lens of ratsadministered cisplatin (A) and before (B), simultaneously (C) and after (D) with cisplatin. (mean± S.D., n = 8). (a) P < 0.05 and (b)P < 0.01 as compared with group control. (d) P < 0.05 and (e) P < 0.01 as compared with group A.

ing effect in the kidney, liver and lens. The highestvitamin E level in the kidney was in the B group.

There were significant differences in the�-caroteneand vitamin A concentrations in the liver and kidneyof the control and A group.�- carotene concentrationsin the liver and kidney were significantly (P < 0.05andP < 0.01) lower in the group A than in the controlgroup. In the group A, liver vitamin A concentrationwas significantly (P < 0.05) lower than in the con-trol group although there was no significant differencein kidney vitamin A concentration The decreased�-carotene and vitamin A concentrations were signifi-

cantly (P < 0.05 toP < 0.001) increased in the SeVEadministered groups.

4. Discussion

Different strategies have been proposed to inhibitcisplatin-induced toxicity. Development of therapiesto prevent the action of generation of free radicalsmay influence the progression of oxidative renal dam-age, along with the appearance of acute renal dam-age induced by cisplatin. Natural antioxidants such as


vitamin E and selenium can be easily and safety in-creased in tissues by intraperitoneal supplementationsince ideal way such as before, simultaneously andafter with cisplatin is known. In this study, we investi-gated the effects of SeVE on rats treated with before,simultaneously and after with cisplatin.

Although the exact mechanism of cisplatin-inducednephrotoxicity is not well understood, several stud-ies have suggested the involvement of lipid peroxi-dation and free radical formation. Cisplatin generatesROS such as superoxide anion and hydroxyl radicals,and stimulates renal lipid peroxidation (Halliwell andGutteridge, 1999; Weijl et al., 1997). It is accepted thatboth correlate to oxidative stress and cause an imbal-ance between the generation of oxygen derived rad-icals and the organism’s antioxidant potential. It hasbeen shown in various studies that cisplatin admin-istrations are associated with increased formation offree radicals, and with heavy oxidative stress (Weijlet al., 1997; Antunes and Darin, 2000; Antunes et al.,2001; Mora et al., 2003). As a result of an increasein the formation of free radicals in cisplatin-inducedtoxicity, the balance normally present in cells betweenradical formation and protection against them is dis-turbed (Kuhlmann et al., 1997; Conklin, 2000). Thiswill lead to oxidative damage of cell components,e.g. proteins, lipids, and nucleic acids (Packer andLandvik, 1990).

Some investigators have reported an associationbetween MDA and cisplatin-induced complications(Kuhlmann et al., 1997; Antunes and Darin, 2000;Silva et al., 2001) while other (Mora et al., 2003) refutethis. A cause of the different reports, cisplatin-inducedtoxicity in each case was associated with elevated lev-els of different indirect measurement of lipid peroxi-dation. Our findings show that MDA levels in the liver,lens and kidney of the A group were higher than inthe control group (Fig. 1). A possible explanation forthe enhancement of MDA concentration may be de-creased formation of antioxidants in cisplatin-inducedtissues, which in view of augmented activity of ROSallows a consequent increase in MDA production.

In various studies which investigated the role of ox-idant stress in the kidney, lipid peroxides are reportedto be increased (Kuhlmann et al., 1997; Antunes andDarin, 2000). On the other hand, an insufficiency in en-zymatic and non enzymatic antioxidant systems withcancer is also reported (Combs and Combs, 1986;

Weijl et al., 1997). Like these reports (Combs andCombs, 1986; Weijl et al., 1997; Kuhlmann et al.,1997; Antunes and Darin, 2000), GSH-Px in the lensand kidney were lower in the cisplatin injected animalsthan in the control animals although the MDA lev-els were increased. Several physio-pathogenic mecha-nisms have been proposed to explain increased MDAand decreased antioxidant levels in the cisplatin toxic-ity. Meanwhile, two distinct pathophysiological mech-anisms have been recognized as primary promoters ofcellular damage, i.e. inhibition of protein synthesis andglutathione protein depletions; in contrast, mitochon-drial damage, inhibition of membranous transport pro-teins and lipid peroxidation are considered merely assequelae of established cell damage (Kuhlmann et al.,1997).

The decreased liver and lens�-carotene and vita-min E concentration found in rats treated with cis-platin rats compared to control group rats indicate thatlipid-phase antioxidant defense is impaired duringcisplatin treatment. In similar to our results, a numberof studies have reported that vitamin E concentrationin plasma or tissue from cancer patients and ani-mals treated with cisplatin.Elsendoorn et al. (2001)reported no changes in vitamin E levels whereasothers have reported decreases (Bogin et al., 1994;Appenroth et al., 1997; Iqbal et al., 1998; Halliwelland Gutteridge, 1999). In this study,�-carotene con-centrations in the kidney and liver and vitamin Econcentrations in the lens and liver of rats treated withcisplatin had significantly lower compared to controlgroup rats. These findings are similar to the results ofother investigators studying antioxidants vitamins inrelation to risk factors in subjects and animals treatedwith cisplatin (Bogin et al., 1994; Halliwell andGutteridge, 1999; Appenroth et al., 1997; Iqbal et al.,1998; Silva et al., 2001).

Vitamin E concentration in the kidney was signif-icantly higher in animals treated with cisplatin thanin control group animals. There is a possible expla-nation for the higher vitamin E concentration in an-imals treated with cisplatin than in control group. Itis that the increased�-tocopherol concentrations inanimals treated with cisplatin were due to upregula-tion of the�-tocopherol transfer protein in response toincreased oxidative stress. Although this mechanismhas not been well elucidated,Tamai et al. (2000)re-ported upregulation of the transfer protein in response


to increased oxidative stress associated with diabetesmellitus. The observed increase of vitamin E in thekidney vitamin E is consistent with the findings ofother investigators studying response of plasma vita-min E to cisplatin administration, where an increasein post cisplatin administration plasma�-tocopherolconcentrations was reported (Bogin et al., 1994; Iqbalet al., 1998; Halliwell and Gutteridge, 1999).

The selenium-containing enzyme GSH-Px protectscells against ROS. In this study, decreased activities ofthe key antioxidant, GSH-Px, were found in the kid-ney, liver and lens of rats treated with cisplatin. Thisresult indicates that the increase in MDA in the kidney,lens and liver of rats treated with cisplatin may be re-lated to the decrease in the activity of GSH-Px, whichscavenges hydroperoxides and lipid peroxides. In thisstudy, administration of SeVE exhibits GSH-Px-likeactivity which prevented the decrease of GSH-Px ac-tivity in the kidney, lens and liver.Husain et al. (1998)andYoshida et al. (2000)showed that the reduction ofGSH-Px activity in the renal tissue of rats treated withcisplatin alone was prevented by co-treatment with eb-selen, which is a selenium compound.

One of the most important intracellular antioxidantsystem is the glutathione redox cycle. Glutathione isone of the essential compounds for maintaining cellintegrity because of its reducing properties and partici-pation in the cell metabolism (Conklin, 2000). The ex-act mechanisms of the cisplatin-induced changes in re-nal GSH concentration are not completely elucidated.Thus, glutathione may modulate metal reduction, andthe thiol portion is very reactive with several chem-ical compounds, mainly with alkylating agents suchas cisplatin (Halliwell and Gutteridge, 1999). In thisstudy, GSH levels in the kidney of rats treated withcisplatin were significantly differed compared to con-trol group rats. These findings are similar to resultsof other investigators studying antioxidants vitaminsin relation to risk factors in diabetic subjects and ani-mals treated with cisplatin (Halliwell and Gutteridge,1999; Antunes and Darin, 2000; Caffrey and Frenkel,2000; Silva et al., 2001).

In conclusion, we have shown that experimental cis-platin administration is a consideration with increasedlipid peroxidation in rats. We, therefore, suggest thatoxidative stress is a cause in the cisplatin inducedpathophysiology. Simultaneously, we have studied theco-administration of SeVE as an approach to ame-

liorate cisplatin-induced nephrotoxicity. These agentsmay protect against cisplatin-induced toxicity by inhi-bition of the inactivation of glutathione and antioxidantsystem by cisplatin, upregulation of GSH-Px levelsand vitamin E in the kidney. The moderate seleniumand high dose vitamin E supplementations may playa role in cisplatin induced nephropathy and cataractformation of cancer patient.

References

Antunes, L.M.G., Darin, J.D.C., Bianchi, M. de L.P., 2000.Pharmacol. Res. 41, 405–411.

Antunes, L.M.G., Darin, J.D.C., Bianchi, M. de L.P., 2001.Pharmacol. Res. 43, 145–150.

Appenroth, D., Fröb, S., Kersten, L., Splinter, E.K., Winnefeld,K., 1997. Protective effects of vitamin C and E on cisplatinnephrotoxicity in developing rats. Arch. Toxicol. 71, 677–683.

Bogin, E., Marom, M., Changes in serum, Y., 1994. Liver andkidneys of cisplatin-treated rats effects of antioxidants. Eur. J.Clin. Chem. Clin. Biochem. 32, 843–851.

Caffrey, P.B., Frenkel, G.D., 2000. Selenium compounds preventthe induction of drug resistance by cisplatin in human ovariantumor xenograftsin vivo. Cancer Chemother. Pharmacol. 46,74–78.

Combs, G.F., Combs, B.S., 1986. The role of selenium in nutrition.Academic Press Inc., London, pp. 206–312.

Conklin, K.A., 2000. Dietary antioxidants during cancerchemotherapy: impact on chemotherapeutic effectiveness anddevelopment of side effects. Nutr. Cancer 37, 1–18.

Czernichow, S., Hercberg, S., 2001. Interventional studiesconcerning the role of antioxidant vitamins in cardiovasculardiseases: a review. J. Nutr. Health Aging 5, 188–195.

Desai, I.D., 1984. Vitamin E analysis methods for animal tissues.Methods Enzymol. 105, 138–147.

Elsendoorn, T.J., Weijl, N.L., Mithoe, S., ZwindermanVan Dam,A.H.F., De Zwarf, F.A., et al., 2001. Chemotherapy-inducedchromosomal damage in peripheral blood layphocytes of cancerpatients supplemented with antioxidants or placebo. Mutat.Res. 498, 145–158.

Halliwell, B., Gutteridge, J.M.C., 1999. Lipid peroxidation: aradical chain reaction. In: Halliwell, B., Gutteridge, J.M.C.(Eds.), Free radicals in biology and medicine. Clarendon Press,Oxford, UK, 1999, pp. 188–276.

Husain, K., Morris, C., Whitworth, C., Trammell, G.L., Rybak,L.P., Somani, S.M., 1998. Protection by ebselen againstcisplatin-induced nephrotoxicity: antioxidant system. Mol. Cell.Biochem. 178, 127–133.

Iqbal, M., Rezazadeh, H., Ansar, S., Athar, M., 1998.�-Toco-pherol (vitamin E) ameliorates ferric nitriloacetate (Fe-NTA)-dependent renal proliferative response and toxicity diminutionof oxidative stress. Hum. Exp. Toxicol. 17, 163–171.

Kuhlmann, M.K., Burkhardt, G., Köhler, H., 1997. Insights intopotential cellular mechanisms of cisplatin nephrotoxicity and


their clinical application. Nephrol. Dial. Transplant. 12, 2478–2480.

Lawrence, R.A., Burk, R.F., 1976. Glutathione peroxidase activityin selenium-deficient rat liver. Biochem. Biophys. Res.Commun. 71, 952–958.

Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J., 1951.Protein measurement with the Folin-Phenol reagent. J. Biol.Chem. 193, 265–275.

Mora, L. de O., Antunes, L.M.G., Franceskocato, H.D.C.,Bianchi, M. de L.P., 2003. The effects of oral glutamine oncisplatin-induced nephrotoxicity in rats. Pharmacol. Res. 47,511–522.

Nazıroglu, M., Çay, M., 2001. Protective role of intraperitoneallyadministered vitamin E and selenium on the antioxidativedefense mechanism in rats with diabetes inducedstreptozotocin. Biol. Trace Elem. Res. 79, 149–159.

Nazıroglu, M., 2003. Enhanced testicular antioxidant capacity instreptozotocin induced diabetic rats: protective role of vitaminsC, E and selenium. Biol. Trace Elem. Res. 94, 61–71.

Packer, L., Landvik, S., 1990. Vitamin E in biological systems.In: Emerit, I., Packer, L., Auclair, C. (Eds). Antioxidants intherapy and preventive medicine. Plenum Press, New York,pp. 93–104.

Placer, Z.A., Cushman, L., Johnson, B.C., 1966. Estimationof products of lipid peroxidation (malonyl dialdehyde) inbiological fluids. Anal. Biochem. 16, 359–364.

Sedlak, J., Lindsay, R.H.C., 1968. Estimation of total, proteinbound and non-protein sulfhydryl groups in tissue withEllmann’s reagent. Anal. Biochem. 25, 192–205.

Silva, C.R., Antunes, L.M.G., Bianchi, M. de L.P., 2001.Antioxidant action of bixin against cisplatin-induced chrosomeaberrations and lipid peroxidation in rats. Pharmacol. Res. 43,561–566.

Suzuki, J., Katoh, N., 1990. A simple and cheap method formeasuring vitamin A in cattle using only a spectrophotometer.Jpn. J. Vet. Sci. 52, 1282–1284.

Tamai, H., Kim, H.S., Hozumi, M., Kuno, T., Murata, T.,Morinobu, T., 2000. Plasma�-tocopherol level in diabetesmellitus. Biofactors 11, 7–9.

Weijl, N.I., Cleton, F.J., Osanto, S., 1997. Free radicals andantioxidants in chemotherapy-induced toxicity. Cancer Treat.Rev. 23, 209–240.

Yoshida, M., Itzuka, K., Hara, M., Nishijima, H., Shimada, A.,Nakada, K., Satoh, Y., Akama, Y., Terada, A., 2000. Preventionof nephrotoxicity of cisplatin by repeated oral administrationof ebselen in rats. Tohoku J. Exp. Med. 191, 209–220.

Selenium and high dose vitamin E administration protects cisplatin-induced oxidative damage to...

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