Tissue and Cell 46 (2014) 170177

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Review

Effect of Sodium Selenite and Vitamin E on the ReAn Ultrastructure Study

Adel M. HAnatomy Depa

a r t i c l

Article history:Received 5 NoReceived in reAccepted 13 MAvailable onlin

Keywords:Sodium seleniVitamin EBax proteinUltrastructure

in E tps. G, resp

for 2ally. oupspsul

ical stlatioith los disc

selenite-induced changes in cortical tissues. Clinically, it is advised to add vitamin E to avoid seleniumoverdose hazards.

2014 Elsevier Ltd. All rights reserved.

1. Introdu

Seleniumantioxidantcance in varHumans ansoilplantof the minevitamin E istive role aga2004).

Vitaminantioxidantselenium, athe free rad2004). Thusother (Tiwa

CorresponAnatomy DepaSaudi Arabia. T

E-mail add(Mustafa H.N.)

http://dx.doi.o0040-8166/ ction

is a critical trace element for human health, potent, and increasingly recognized for its clinical signi-ious pathophysiologic conditions (Tos-Luty et al., 2003).d domestic animals acquire selenium mainly from thewater system in which plants are an immediate sourceral (Tian et al., 2005). As a chain breaking, soluble lipid,

an efcient antioxidant that has a well-known protec-inst a wide range of human diseases (Niki and Noguchi,

E and selenium complement each other in their role ass. Vitamin E inhibits free radical production, whereass a constituent of glutathione peroxidase, neutralizesicals present in the biological matrix (Naziroglu et al.,, vitamin E and selenium have a sparing effect on eachry et al., 2006). Vitamin E is thought to be involved

ding author at: King Abdulaziz University, Faculty of Medicine,rtment, Ground Floor Building No. 8, P.O. Box: 80205, Jeddah 21589,el.: +966 566 764 762.resses: hesham977@hotmail.com, hesham977@yahoo.com.

in an unlimited range of physiological and biochemical functions.The molecular mechanism of these functions could be mediated byeither antioxidant activity or by its action as a membrane stabilizer,as its basic function is to protect all membrane lipids and unsatu-rated fatty acids against oxidative degradation (Wang and Quinn,2000).

Daily required selenium for adults is 200 g/day. The upperlimit of the tolerable intake of selenium is 400 g/day, as deter-mined by the Food and Nutrition Board of the Institute of Medicinebased on the avoidance of hair and nail brittleness and loss and ini-tial signs of chronic selenium toxicity (Nuttall, 2006). The toxicityoccurs when a human is exposed to excess consumption, overdosethrough selenium medications, or excessive and unregulated useof supplementation. The most common reported symptoms aregastrointestinal disturbances, garlic odor in the breath, lethargy,irritability, and nervous irregularities. However, extreme cases canlead to hepatic cirrhosis, edema of the lung, and death (Reid et al.,2004). Selenium can exert a pathogenic effect either in deciencyor in excess (i.e., excess of this chemical may lead to changes similarto those observed in its deciency). These changes are caused byfree radical degeneration, inhibition of synthesis of proteins, andintensication of lipid peroxidation (El-Demerdash, 2004). Inor-ganic selenium as sodium selenite (Na2SeO3) is frequently used,along with vitamin E, for supplementation in animals identied

rg/10.1016/j.tice.2014.03.0022014 Elsevier Ltd. All rights reserved.ussein, Hamid A. Saleh, Mustafa H.N.

rtment, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia

e i n f o

vember 2013vised form 13 March 2014arch 2014e 20 March 2014

te

a b s t r a c t

This study examined the use of vitamrats (n = 50) was divided into ve grouselenite (2 mg/kg) for 2 and 4 weeks(2 mg/kg) and vitamin E (100 mg/kg)anti-BCL2 and examined ultrastructurpositive after 4 weeks and nearly all grstudy revealed lesions in Bowmans carevealed damage and necrosis of cortlular changes were seen, such as vacuospace scattered through the cortex wand cellular rupture. Present ndingnal Cortex in Rats:

o alleviate toxic effects of sodium selenite. Adult male albinoroup 1 was control, Groups 2 and 4 were treated with sodiumectively, Groups 3 and 5 were treated with sodium selenite

and 4 weeks, respectively. Renal tissues were studied usingPositive Bax immunoreactivity was detected after 2 and more

improved with co-administration of vitamin E. Ultrastructurale and proximal convoluted tubules. The submicroscopic studyructures after 2 and 4 weeks, respectively. After 4 weeks, cel-n and moderate degeneration of cells, widening of the urinaryss of cellular details, formation of apical buds, degeneration,losed an ameliorative effect of adding vitamin E to sodium

A.M. Hussein et al. / Tissue and Cell 46 (2014) 170177 171

with selenium deciency or in animals living in selenium decientareas (Hemingway, 2003).

The toxic outcomes of selenium in vitro, despite being non-specic, are avoided by the presence of vitamin E (Beyrouty andChan, 2006peroxidatio2003). Thershould be tof selenium

2. Materia

2.1. Chemic

All chemselenite (Nafrom Sigma

2.2. Animal

Adult mobtained froUniversity. ative humidroom (25 the laboratallowed frelibitum.

2.3. Experim

The animcages of tetreatment sdaily dose were givenbody weigh4 weeks, rof sodium 0.5 ml distildissolved intively.

Sodium (100 mg/kgously publisof vitamin Eicity (El-Dethe morninmals. At 2 athe kidneyshistopathol

King AbBiomedicalprotocol. Thmals in sciHealth (NIHreported heAbdulaziz Uexperiment

2.4. Histolo

Renal saxed in 10grades of a

sections (5 m thick) were stained with hematoxylin and eosinstain for routine histological inspection (Ibrahim et al., 2011).The sections were viewed and photographed with an Olympuslight microscope (Olympus BX53, Tokyo, Japan) and an attached

raph werngh

mun

rexposis inatlonaribetionsor 1 t thaationrdizare co

jar secu

The a and essurthe smeras repure.

ensing wernobeA) wtissudy u5, Bi3 drmidinkedted fereas r

oxylixam

preconte

ectro

al sa gluempbuffeehydide,thicklue. omeds. T

citricros). Supra-nutritional doses of vitamin E can reduce lipidn and mortality in selenite-treated animals (Saito et al.,efore, the aim of this study is to determine if vitamin Eaken daily alongside selenium to alleviate the toxicity

overdose.

l and methods

als

icals, including vitamin E (-tocopherol) and sodium-selenite: Na2SeO3) used in this study were purchasedAldrich Chemical Co. (St. Louis, MO, USA).

s

ale Wistar rats (n = 50), weighing 200225 g, werem the Experimental Animal Laboratory, King AbdulazizThey were housed under standard lightening and rel-ity (50 15%) conditions in a temperature-controlled

2 C). The animals were allowed to acclimatize toory environment for 10 days, and then they weree access to standard dry pellet diet and water ad

ental design

als were randomly separated into ve polypropylenen individuals each and were administered 0.5 ml ofolution via oral gavage. Group 1 (control) received aof corn oil for a period of 4 weeks. Groups 2 and 4

daily doses of sodium selenite at a dose of 2 mg/kgt diluted in 0.5 ml distilled water for periods of 2 andespectively. Groups 3 and 5 were given daily dosesselenite at a dose of 2 mg/kg body weight diluted inled water and 0.5 ml vitamin E (100 mg/kg body weight)

corn oil orally for periods of 2 and 4 weeks, respec-

selenite (2 mg/kg body weight) and vitamin E body weight) doses were chosen according to previ-hed data (Turan et al., 2003; Yilmaz, 2005) and the dose

was previously shown to be effective in lowering tox-merdash, 2004). The chemicals were administered ing (between 8:00 AM and 9:00 AM) to non-fasted ani-nd 4 weeks after treatment, the rats were sacriced and

were removed, collected and weighed, and sampled forogical examination.dulaziz Universitys Faculty of Medicine Hospital

and Research Ethics Committee approved the studye general guidelines for the use and care of living ani-entic investigations from the National Institutes of

Publication No. 80-23) were followed. All experimentsre complied with current laws and regulations of Kingniversity, Saudi Arabia, on the care and handling ofal animals.

gical and morphometric evaluation

mples were dissected longitudinally. Then they were% neutral buffered formalin, dehydrated in ascendinglcohol, and embedded in parafn wax. Next, parafn

photogImagesics, Sha

2.5. Im

Ove(apoptdetermmonocas desc

Secoven fproducrehydrstandaing moCoplinjar was2013).120 Cthe prallow and imstep wproced

TheimmerThe Po(DiamiCA, USin the antiboorb465and 2the huHRP-liincubadrops wstain whematwere ebrown(Albam

2.6. El

Renin 2.5%room tphate were dlene ox(1 m idine b(Ultratper griby leadtron m2012). machine (Olympus E-330, Olympus Optical Co. Ltd.).e analyzed using Image-Pro Plus v4.5 (Media Cybernet-ai, China).

ohistochemistry using an anti-Bax antibody

ression of Bax protein accelerates apoptotic cell deathinducers). Immunohistochemical detection of Bax andion of the expression level were achieved using a mousel anti-Bax antibody and avidinbiotin complex stainingd by the manufacturer.

were cut into 4 m slices and then xed in a 65 Ch. Trilogy (Cell Marque, CA, USA, Cat# 920p-06) is at combines the 3 pretreatment steps: deparafnization,, and antigen unmasking. Using this product enhancestion of the pretreatment procedure, thereby, produc-nsistent and reliable results. The slides were placed in alled with 200 ml of the trilogy working solution, and therely positioned in an autoclave (Morsy and El-Moselhy,utoclave was adjusted so that the temperature reachedmaintained this temperature for 15 min, after whiche was released and the Coplin jar was removed tolides to cool for 30 min. Sections were then washedsed in Tris-buffered saline (TBS) to adjust the pH; thiseated between each step of the immunohistochemistry

dogenous peroxidase activity was determined bythe slides in 3% hydrogen peroxide for 10 min.

-StainTM 1.0 Poly HRP (Horseradish Peroxidase) DABnzidine) Kit Cat# 54-0017 (Genemed Biotechnologies,as used to visualize any antigen-antibody reactiones (for qualitative detection of antigen). The primarysed for Bax immunohistochemical staining (Bax Cat #orbyt, Cambridge, United Kingdom) was diluted 1:500,ops were applied. The slides were then incubated inty chamber overnight at 4 C. Subsequently, polyclonal

antibody conjugates were applied to each slide andor 20 min. The DAB chromogen was prepared, and 23

applied to each slide and incubated for 2 min. The DABinsed off, counterstaining was performed with Mayern, and then a cover slip was attached before the slidesined under a light microscope. The cells that displayedipitation were considered positive for Bax expression

et al., 2013).

n microscopy

mples were cut into small pieces (1 mm3) and xedtaraldehyde (pH 7.4) in phosphate buffer for 2 h aterature. Post-xation was performed in the same phos-r containing 1% osmium tetroxide (OsO4). The tissuesrated in graded ethanol solutions, transferred to propy-

and nally embedded in Epon 812. Semithin sections) were cut using a glass knife and stained with tolu-Ultrathin sections were obtained using LKB ultratome

NOVA, LKB 2188, Bromma, Sweden) and spread on cop-he sections were stained with uranyl acetate followedate and examined at 80 kV with a transmission elec-cope (JEM-2000EX; JEOL, Tokyo, Japan) (Hirose et al.,

172 A.M. Hussein et al. / Tissue and Cell 46 (2014) 170177

Table 1Body weights of rats treated with sodium selenite.

Treatment duration Body weight (g) Kidney weight (g)

Control group(n = 10)

Sodium selenitegroup (n = 10)

Sodium seleniteand vitamin Egroup (n = 10)

Control group(n = 10)

Sodium selenitegroup (n = 10)

Sodium seleniteand vitamin Egroup (n = 10)

1st day 196.10 1.101 196.10 1.101 196.10 1.101 2 weeks 199.40 0.699 194.50 0.527 199.00 0.816 0.628 0.002 0.619 0.001 0.626 0.003

P < 0.0001 P < 0.207 P < 0.0001 P < 0.1284 weeks 209.50 0.527 204.10 0.876 208.60 0.516 0.648 0.002 0.639 0.001 0.646 0.003

P < 0.0001 P < 0.005 P < 0.0001 P < 0.128

Values are expressed as a mean standard deviation. The analysis was made using one-way ANOVA test (LSD).

Table 2Comparison of the mean values of kidney histological parameters (m) of the experimental groups with the control group.

Groups Renal corpuscle Urinary space PCT

Control group 82.96 7.83 9.54 2.26 39.28 3.24Group receiving sodium selenite for 2 weeks 65.59 8.04 **P < 0.01 7.11 2.28 35.11 2.89Group receiving sodium selenite for 4 weeks 54.66 9.33 ***P < 0.001 7.4 2.35 18.63 1.66 ***P < 0.001Group receiving sodium selenite + vitamin E for 2 weeks 70.52 12.70 *P < 0.05 6.34 1.47 **P < 0.01 36.3 3.49Group receiving sodium selenite + vitamin E for 4 weeks 60.81 9.34 *P < 0.05 5.64 1.54 ***P < 0.001 21.85 2.78 ***P < 0.001

Values are expressed as a mean standard deviation. *P < 0.05 differences signicant. **P < 0.01 differences considerably signicant. ***P < 0.001 difference very signicant.For statistical signicance, experimental groups have been compared to their respective control group.

2.7. Statistical analysis

Data are expressed as a mean standard deviation. Statisticalanalysis of all data was performed using SPSS for Windows (Ver-sion 22; SPSS Inc., Chicago, IL, USA). One-way analysis of variance(ANOVA) was used, followed by post hoc analysis using LSD test.Values were considered statistically signicant when the p-valuewas less than 0.05.

3. Results

3.1. Effect of sodium selenite on rat general health

In sodium selenite-treated groups, few clinical signs, such asreduced activity, cumulative weakness, petty diarrhea, and hairloss, were observed. Death was not observed in any of the experi-mental groups during the treatment period (4 weeks).

Fig. 1. Immunin the kidney selenite 2 mg/ochemical Bax staining. (A) Positive Bax immunoreactivity in the kidney of the groups treagroups treated with sodium selenite 2 mg/2 week and vitamin E. (C) Strong positive Bax4 week. (D) Weak Bax immunoreactivity in kidney groups treated with sodium selenite 2ted with sodium selenite 2 mg/2 week. (B) Faint Bax immunoreactivity immunoreactivity in the kidney of the groups treated with sodium

mg/4 week and vitamin E. (Bax immunostaining 200).

A.M. Hussein et al. / Tissue and Cell 46 (2014) 170177 173

Fig. 2. (A) An electron micrograph of a section of the group treated with sodium selenite 2 mg/2 week showing the Bowmans capsule of the glomeruli formed by hypertrophiedbasal lamina (BL) with narrowing of the urinary space and attened, elongated parietal epithelium (N). The urinary space is narrow or nearly absent (thin black arrows), andthe capillary tuft is moderately congested Red Blood Corpuscles (RBC). Note: The visceral epithelium shows dentate nucleus (DN). (B) An electron micrograph of a section ofthe kidney of the group treated with sodium selenite 2 mg/kg and vitamin E showing the glomerular tuft with apparent congestion of the capillaries (RBC) and the Bowmanscapsule showing thickening of the basal lamina (BL) with improved urinary space narrowing and hypertrophy of the podocyte. Note: The visceral epithelium shows dentatenucleus (DN). (C) An electron micrograph of a section of the group treated with sodium selenite 2 mg/4 week showing the Bowmans capsule greatly thickened basal lamina(thick white arrows) and parietal epithelium at and elongated (N). The urinary space is greatly reduced or nearly absent (thin white arrows) and the capillary tuft (RBC) iscongested with obvious moderate thickening of the basal lamina (BL) and hypertrophy of the podocyte. Note: The visceral epithelium shows dentate nucleus (DN). (D) Anelectron micrograph of a section of the group treated with sodium selenite 2 mg/4 week showing dilation of the capillary tuft (CT) and lled with plasma and the shrinkageand atrophy of the podocyte (Pod). Shrinkage and condensation of the cytoplasm of the parietal epithelium (PE) includes thickening of the Bowmans capsule basal lamina(BL), narrowing of the urinary space (US), and numerous invaginated corrugated membrane proles (C) at the base. (E) An electron micrograph of a section of the grouptreated with sodium selenite 2 mg/4 week and vitamin E showing the Bowmans capsule with apparently moderate thickening of the basal lamina (BL) with seeminglyimprovement of the urinary space narrowing and less congestion of the capillary tuft (RBC). The size of the mesangial cell and matrix are reduced.

174 A.M. Hussein et al. / Tissue and Cell 46 (2014) 170177

A.M. Hussein et al. / Tissue and Cell 46 (2014) 170177 175

3.2. Evaluation of body and kidney weights

After 2 and 4 weeks, the body weights in the sodium selenite-treated groups were signicantly lower than in the controland sodium selenite + vitamin E-treated groups (P < 0.0001 forall). While insignicant differences in body weight were seenbetween the control and sodium selenite + vitamin E-treatedgroups (P < 0.207) after 2 weeks, signicant differences wereseen after 4 weeks (P < 0.005). After 2 and 4 weeks, the kidneyweights in the sodium selenite-treated group were signicantlylower than in the control and sodium selenite + vitamin E-treatedgroups (P < 0.0001 for all); meanwhile, insignicant differenceswere seen between the control and sodium selenite + vitamin E-treated groups (P < 0.128 for both) (Table 1).

3.3. Morphometric results

The kidneys from the control group showed normal structure ofthe cortex. While, the experimental groups kidneys revealed his-tological alterations that were mainly cortical. Renal corpuscle andproximal convoluted tubules are markedly affected while the distalconvoluted tubules showed minimal or no detectable histologicalalterations.in these gro

Renal coin size in th4 weeks (Ptreated witdiminutionspace showwith sodiumweeks (P < 0eter revealefor 4 weeksafter treatm(Table 2).

3.4. Immun

To detereffects of vdetected usBax was mthe intensigroup was augmentednied by ren

reduced renal expression of Bax, which was likely associated withits anti-apoptotic effects. Bax-stained cells were not observed inthe control group, while sodium selenite-treated rats for 2 weekshad dense immune-positive cells (Fig. 1A) and those treated withsodium selenite for 4 weeks showed more dense immune-positivecells (Fig. 1C). The groups exposed to sodium selenite along withvitamin E for 2 or 4 weeks had minimal immune-positive cells(Fig. 1B, D).

3.5. Ultrastructure results

The renal cortex, being a major portion of the kidney, wasstudied with particular regard to the cortical ultrastructuralappearances. Ultrastructural submicroscopic observations of therenal tissues of the control rats disclosed a healthy picture of theBowmans capsule, tubular system, and endothelial cells in theglomerular capillaries with a highly rich and fenestrated endothe-lium with large pores. The epithelium of the proximal renal tubularwas distinguished by a dense brush border, rounded vesicularnucleus near the base or the central portion, endocytic vesiclesin the apical portion, rare lysosomes, and elongated, rounded

ondr cister 2 we wit

spaal co

ant mreoveexpoith imted c

reve abunr 4 we witace

ocyted de

ed mr 4 d Bobasaing al coondr

(Fig

Fig. 3. (A) An week basal lamina ( ontainelectron dense ) is dhaving abunda . (B) Aselenite 2 mg/ ule. Twith preserved nd seecontains large cell s(BL) is thin wi graphrevealing the t a (BL)characteristic bundais all over the agniweek revealin lar epapparent nucle d, andmitochondria up tretubular epithe ted nofew electron d ucleucell surface ha ickenbase of the cel In addition, the medulla showed minimal or no changesups.rpuscle diameter showed highly signicant reductionose treated with sodium selenite for 2 (P < 0.001) and

< 0.0001) compared to the control. Moreover, thoseh sodium selenite and vitamin E showed signicant

in size compared to the control while the urinaryed highly signicant improvement with those treated

selenite and vitamin E for 2 weeks (P < 0.001) and 4.0001). In addition, proximal convoluted tubular diam-d diminution in diameter after sodium selenite alone

(P < 0.0001) with apparent signicant improvementsent with sodium selenite and vitamin E (P < 0.0001)

ohistochemical results

mine the mechanism underlying the anti-apoptoticitamin E, Bax (a marker protein of apoptosis) wasing immunohistochemistry. The results revealed thatainly expressed in the cytoplasm of renal cells, andty of Bax expression in the sodium selenite-treatedhigher than that in the control group. Sodium selenite

renal expression of Bax, which was probably accompa-al apoptosis. Moreover, co-administration of vitamin E

mitochculum

Aftecapsulurinaryproximabund

Mogroup ina, wcongestubulecristae

Aftecapsulnary spof podrevealeclump

Afteexposeof the narrowproximmitochlamina

electron micrograph of a section of the group treated with sodium selenite 2 mg/2 BL) is thin with numerous invaginated membranes in the cell cytoplasm. The cell c

rounded bodies varying in size that seem to be lipid components (L). The nucleus (Nnt, closely packed microvilli (Vi) and congestion of the peritubular capillary (RBC)2 week and vitamin E revealing the tubular epithelium of proximal convoluted tub

cristae and an overall decrease in electron dense rounded bodies that vary in size a nucleolus (Nu) nuclei, and regains the characteristic appearance of chromatin. Theth numerous invaginated membranes in the cell cytoplasm. (C) An electron microubular epithelium of proximal convoluted tubule with thickening of the basal laminelectron density of the nuclear chromatin, the mitochondria (M) is obviously less acytoplasm, and the surface microvilli (Vi) is clumped and not straight. (D) Higher mg the tubular epithelium of proximal convoluted tubule. The nucleus (N) of the tubuolus (Nu), the myelin like gure (MF) and the surface microvilli (Vi) are long, clumpe(M) and apparent vacuoles (V). (E) An electron micrograph of a section of the grolium of proximal convoluted tubule. The cell contains abundant pleomorphic elongaense rounded bodies varying in size that seem to be of lipid components (L). The ns abundant, closely packed long microvilli (Vi). The basal lamina (BL) is sparingly thl.ia with preserved cristae and rough endoplasmic reti-rnae placed between the mitochondria.eeks, sodium selenite treated group exposed Bowmansh hypertrophy of the basal lamina, narrowing of thece with congested capillaries. While the epithelium ofnvoluted tubule revealed pleomorphic mitochondria,icrovilli and thin basal lamina (Figs. 2A and 3A).r, after 2 weeks, sodium selenite and vitamin E treatedsed Bowmans capsule with thickening of the basal lam-provement of the urinary space narrowing and less

apillaries. While the epithelium of proximal convolutedaled apparently healthy mitochondria with preserveddant microvilli and thin basal lamina (Figs. 2B and 3B).eeks, sodium selenite treated group exposed Bowmansh greatly thickened basal lamina, markedly reduced uri-with congested capillaries and shrinkage and atrophys. While the epithelium of proximal convoluted tubulegenerated ballooned mitochondria with lost cristae,icrovilli and thick basal lamina (Figs. 2C, D and 3C, D).weeks, sodium selenite and vitamin E treated groupwmans capsule with apparently moderate thickeningl lamina, seemingly improvement of the urinary spaceand less congested capillaries. While the epithelium ofnvoluted tubule revealed noticeably healthy unaffectedia, plentiful microvilli and sparingly thickened basals. 2E and 3E).

revealing the tubular epithelium of proximal convoluted tubule. Thes abundant pleomorphic elongated mitochondria (M) and numerousentated, vesicular, and contain a large nucleolus (Nu). The cell surfacen electron micrograph of a section of the group treated with sodiumhe cell contains abundant pleomorphic elongated mitochondria (M)m to be of lipid components (L). The nucleus (N) is rounded, vesicular,urface has abundant, closely packed microvilli (Vi). The basal lamina

of a section of the group treated with sodium selenite 2 mg/4 week of the tubule. The nucleus (N) of the tubular epithelial cell has lost itsnt, vacuolated, lost cristae and more electron dense, vacuolation (V)cation of a section of the group treated with sodium selenite 2 mg/4ithelium has lost its electron density of the nuclear chromatin and an

not straight as usual, and there are more electron dense pleomorphicated with sodium selenite 2 mg/4 week and vitamin E revealing theticeably healthy unaffected mitochondria (M) with preserved cristae,s (N) is rounded and vesicular with an apparent nucleolus (Nu). Theed with numerous invaginated corrugated membrane proles at the

176 A.M. Hussein et al. / Tissue and Cell 46 (2014) 170177

4. Discussion

Amongst the necessary human micronutrients, selenium ispeculiar due to its valuable physiological activity and toxicity. Itmay have anat concentrcarcinogeni

The currweight andreported thtionship bediminishedthe secretiogrowth fact(Wang et aweight of tfound to beative impro2 weeks anagrees withselenite (To

Moreoveeffects of soThese effeccapsule, wieters of proThese resulweeks. Thewith the corelatively imvoluted tubfaint or no e

In our treated groHowever, csion of Bax results indiof Bax.

Previousinhibited ththe level o(GSH) and blocked theand the actgations havapoptosis oour study, vinduced by injuries indof oxidativeapoptosis.

On the uvitamin E, ing of the the urinaryshowed mowith seemi4 weeks.

The ceobserved inKhattab (2in the formappearancedegeneratioations in ththe tubules

damage of the brush borders with darkly stained pyknotic nuclei.Swollen microvilli with many vesicles and swollen, condensedmitochondria with dark matrices and indistinguishable cristaewere observed (Khattab, 2007). Other authors also observed

ion ancet al., oteince o

sel

late sel

cellsviou

lysoraterophodiesed m

ranee au

terst2007brushr resurtex

and furia ry to

In aethyney,r cellitial

of th et a

stude. Tht al., res aed teort

clus

hist caustionamin ent

sel in a

exac

rs co

autho. All

t of

authti-carcinogenic properties at low concentrations, whileations higher than those needed for nutrition, it can bec and genotoxic (Valdiglesias et al., 2010).ent study suggests signicant correlation between body

selenium intake. Sakr and El-Abd (2009) previouslyese ndings in which they found a proportional rela-tween selenium intake and body weight reduction with

food intake. The inhibitory effects of sodium selenite onn of growth hormone and somatomedin C (insulin-likeor 1-binding proteins) may explain growth retardationl., 2007). In addition, a signicant decrease in kidneyhe selenium-treated animals was seen and this was

related to the duration of treatment. There was rel-vement with the co-administration of vitamin E afterd more amelioration was seen after 4 weeks and this

the study of Tos-Luty who studied the effect of sodiums-Luty et al., 2003).r, the present work denotes that the morphologicaldium selenite on the renal tissues are mainly cortical.ts include a decrease in the diameter of the Bowmansdening of the urinary space, and decrease in the diam-ximal convoluted tubules in comparison to the control.ts appeared after 2 weeks and were dramatic after 4se latter ndings agree with that of Khattab (2007), but-administration of vitamin E, these parameters wereproved. On the contrary, the effects on the distal con-ules were minimal, while the renal medulla showedffects, disagreeing with the ndings of Khattab (2007).ndings, the expression of Bax in the sodium selenite-up was increased compared to the control group.o-administration of vitamin E decreased the expres-compared to the sodium selenite-treated group. Thesecate that vitamin E might inuence the renal expression

ndings concluded that the presence of vitamin Ee formation of reactive oxygen species (ROS), decreasedf lipid peroxide, elevated the level of glutathioneactivities of superoxide dismutase and catalase, and

reduction in the mitochondrial membrane potentialivation of caspase-3 (Meng et al., 2011). Some investi-e indicated that vitamin E protects tissues by causingf injured cells (Reddy Avula and Fernandes, 2000). Initamin E prevented the up-regulation of Bax expressionsodium selenite treatment. This may protect cells fromuced by sodium selenite through the down-regulation

stress and the prevention of mitochondrial-mediated

ltrastructure observation and with the co-addition ofthe Bowmans capsule showed amelioration thicken-basal lamina with improvement of the narrowing of

space after 2 weeks. Moreover, the Bowmans capsulere amelioration of the thickening of the basal laminangly improvement of the urinary space narrowing after

llular and submicroscopic ultrastructure changes the second and fourth week agree with the work of007) that described slight histopathological changes

of cloudy swelling of the tubular cells and smudgy of the renal tubular cells. In addition, vacuolarn and narrowing of the cellular lamina and alter-e nuclear appearance was seen. The epithelial cells of

were partially detached with necrotic changes and

dilatatappearLuty ethe prpresensodium2003).

Thesodiumtubuleand obdenseddegenehypertsion bcontainmemb

Somand inet al., of the Similaney coweeksglucosby inju2007).as dimthe kidtubulainterstborderFujieda

OurselenitLuty emeasuadvancto supp

5. Con

Thecortexproporof vitaof pressodiumdualitylate its

Autho

All equally

Conic

Theof the endoplasmic reticulum and changes in the of the mitochondria, as well as the liposomes (Tos-2003). These ndings were determined to be due to

lipid membranes of those cellular organelles or thef glutathione peroxidase, which can be a site for theenite activity (Naziroglu et al., 2004; Tos-Luty et al.,

r ndings agree with Khattab who studied the effect ofenite at low and high doses. The proximal convoluted

showed coagulative necrosis, electron opaque bodies,sly demolished organelles, except for some heavily con-somes. The cytoplasm of these cells contained manyd mitochondria, some swollen and disrupted and othersied with condensed matrices and electron-dense inclu-. The nuclei of some affected cells showing shrinkageany vacuoles with chromatin clumped at the nuclear

(Khattab, 2007).thors ascribed focal areas of tubular dilatation, atrophy,itial brosis as due to selenium decient diets (Fujieda). Other authors reported sloughing or internalization

border as an effect of toxic materials (Khattab, 2007).lts have been studied on the histology of the rat kid-

after treatment with a selenium decient diet for 112ound that selenium deciency induces protein urea andwith renal calcication, which may be mostly induced

the proximal tubule via oxidative stress (Fujieda et al.,ddition, the inhalation of selenium derivatives, suchl selenide, has been associated with tubular injury of

resulting in swelling and vacuolation of the proximals and showing focal areas of tubular dilation, atrophy,brosis, and sloughing or internalization of the brushe proximal tubules (Cherdwongcharoensuk et al., 2005;l., 2007).y indicates a duration-dependent inuence of sodiumese results agree with other authors ndings (Tos-2003). In addition, more statistics and morphometricre recommended to evaluate the concept and morechniques are recommended as stereological techniques

our ndings.

ion

opathological and ultrastructural changes of the renaled by the oral administration of sodium selenite arete to the duration of its intake. The coadministrationE showed amelioration of these changes. The resultswork afford an improved awareness on the activity ofenite and aid in elucidating the causes underlying thection of it as antioxidant or toxic compound so to regu-t use in nutrition and medicine.

ntributions

rs participated in the design of this work and performed authors read and approved the nal manuscript.

interest statement

ors declare that they have no conict of interest.

A.M. Hussein et al. / Tissue and Cell 46 (2014) 170177 177

Acknowledgements

This study was funded by the Deanship of Scientic Research(DSR), King Abdulaziz University, Jeddah, Saudi Arabia (Grant No.15-140-D1432). The authors, therefore, acknowledge with thanksDSR technical and nancial support.

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Effect of Sodium Selenite and Vitamin E on the Renal Cortex in Rats: An Ultrastructure Study1 Introduction2 Material and methods2.1 Chemicals2.2 Animals2.3 Experimental design2.4 Histological and morphometric evaluation2.5 Immunohistochemistry using an anti-Bax antibody2.6 Electron microscopy2.7 Statistical analysis

3 Results3.1 Effect of sodium selenite on rat general health3.2 Evaluation of body and kidney weights3.3 Morphometric results3.4 Immunohistochemical results3.5 Ultrastructure results

4 Discussion5 ConclusionAuthors contributionsConflict of interest statementAcknowledgementsReferences