JJBS Jordan Journal of Biological Sciences

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1 JJBS Jorn Journl of Biologil Sienes Volume 10, Numer 3,Septemer 2017 ISSN Pges Assessment of Antigenotoxi Effet of Nnoselenium n Metformin on Dieti Rts Aeer H. A El-Rhim *, Omim M A-Elmoneim n Ngl A. Hfiz Cell Biology Deprtment, Ntionl Reserh Center, El Thrir Street, Dokki, Giz, Egypt Reeive Mrh 5, 2017 Revise July 24, 2017 Aepte August 4, 2017 Astrt The present stuy ws onute to investigte the protetive role of selenium nnoprtiles on type-2 ietes mellitus (T2DM) in mle rts trete with metformin rug ompre to the ontrol. Chromosoml errtions, DNA frgmenttion, mironuleus tests s well s omet ssy were rrie out. Rts were ivie into four groups: group1, norml ontrol in whih the rts were reeive norml sline solution, group 2, ontrol ieti group in whih the ieti rts were inue using 45mg/Kg oy weight Streptozotoin (STZ). Group 3, Metformin trete group in whih the ieti rts were trete with stnr orl hypoglyemi gent, metformin (100 mg/kg oy weight (.wt) while, Group 4, selenium nnoprtiles (SeNPs) trete group, in whih the rts were trete with nnoselenium stilize in liposome (0.1 mg/kg. wt) orlly plus metformin. Rts were srifie fter 21 ys following the lst injetion. The results emonstrte tht ietes use inrese in hromosoml errtions, mironuleus formtion, DNA frgmenttion s well s DNA mge using omet ssy, ut these prmeters erese in trete nimls with metformin. In ition, metformin + nnoselenium omine tretment use erese in ll stuie prmeters reltive to metformin lone. In onlusion, it ppere tht SeNPs omine with metformin hve ntiieti effet in experimentl moel of T2DM. This effet oul e srie to their ility in eresing frequenies of hromosoml errtions, eresing the numer of mironuleus s well s eresing the perentges of DNA mge. Thus, SeNPs my enefit in linil purposes espeilly in enhning the effet of T2DM. Keywors: Selenium nnoprtiles, Dietes, Bone mrrow, DNA mge, Rts. 1. Introution Dietes mellitus is the most ommon enorine isese in whih there is n inrese in the level of loo sugr over prolonge perio (hyperglyemi) (Worl Helth Orgniztion (WHO), 2014). There re mny symptoms of this isese, suh s inrese ingestion of foo, inrese thirst, musulr proteolysis, postprnil sikness n mge of mny orgns, suh s eyes, nerves, kineys n hert, these symptoms ffet the humn life s whole. If untrete, ietes uses mny prolems, suh s riovsulr iseses, kiney filure n eye mge (WHO, 2013), ieti ketoiosis n nonketetoti hyperosmolr om (Kithi et l., 2009). There re three types of ietes mellitus: type 1, whih results from filure in the pnres (WHO, 2013). Type 2 (the most ommon type of ietes) in whih the ells o not respon to insulin property, n the numer of ignose ses of this type hs inrese over the pst few ees (Olnsky et l., 2003), n it forms out 90% of the ietes ses (Shi n Frnk, 2014) istriute eqully etween women n men (Vos et l., 2012) n the numer is expete to reh 592 million in 2035 (IDF, 2014). Type 3 is gesttionl ietes n this type ours in pregnnt women who hve ietes history in their fmily (WHO, 2013). The tretment of type 2 ietes epens on using mny rugs, suh s sulfonylures, Insulin, peroxisome prolifertor-tivte reeptor- γ gonists n metformin. Metformin (imethyligunie) is the most esrie orl rug use in the tretment of type 2 ietes (The Amerin Soiety of Helth-System Phrmists, 2016; Mruthur et l., 2016) to ontrol the loo sugr level with or without Insulin in onjuntion with life moifition, suh s weight ontrol, speil iet n physil tivity (Srs et l., 2001). Metformin ereses the resistne of insulin, hepti gluose output n enhnes the uptking of gluose (Biley n Turner, 1996; Niset et l., 2004); these to erese fsting n loo gluose y 20 to 40 perent, eresing of hemogloin n oy weight n inrese high ensity lipoprotein (HDL) (Howlett n Biley, 1999). * Corresponing uthor. e-mil: eerel_rhim@yhoo.om.

2 Jorn Journl of Biologil Sienes. All rights reserve - Volume 10, Numer 3 Selenium (Se) is n importnt element for helth with importnt iologil n iohemil funtions ue to its ntioxint properties (Chunying et l., 2006). Se is si omponent of Selenoprotein whih reues oxitive stress (Rymn, 2012, Ahme et l., 2016). High selenium level in loo plsm hs een orrelte with prevention of severl iseses, suh s ners, riovsulr iseses, musle isorers s well s ietes mellitus (Nvrro-Alro n n Lo pez- Mrtı nez, 2000). The reltion etween selenium n T2DM is ontroversil (Wng et l., 2016). So untrete ietes uses oxitive stress whih les to omplitions of the isese n the ntioxint intke is onsiere s eing fvorle for the therpy of ietes (Mueller et l., 2009), n results in eresing insulin, homeostsis moel of ssessment-insulin resistne, serum high-sensitivity C- retive protein, n inresing the quntittive insulin sensitivity hek inex sore s well s the onentrtion of the totl ntioxint pity (Frrokhin et l., 2016). It is ler tht Selenium plys protetive role ginst T2DM (Steinrenner n Sies, 2009). Furthermore, it hs een foun tht the hlf-life of some therpeutis n e improve y onjugting them to nnorriers (Mkhluf et l., 2008; Liu et l., 2012). It is ovious tht selenium nnoprtiles (SeNPs) hve een onsiere promising tool in rug therpies of type 2 ietes; this is ue to its low toxiity n high therpeuti properties (Ro et l., 2014). SeNPs hve een wiely stuie euse they hve vne iologil tivity n high photoeletri performne, ntioxint tivity, ntiner, n immunesystem enhnements (Go et l., 2002; Gtes et l., 2002). By ompring Se with SeNPs, it ws foun tht SeNPs n serve s potentil hemopreventive gent with reue risk of Se toxiity (Zhng et l., 2005; Wng et l., 2007). The purpose of the present stuy is to investigte the tion of nnoselenium, omine with metformin, on the inution of hromosoml errtions, formtion of mironulei n DNA frgmenttion in streptozotoin inue T2DM rts ompre to those helthy ontrols. 2. Mterils n Methos 2.1. Experimentl Animls For the present stuy, ult femle lino rts of Westr Strin weighing g were otine from the Holing Compny for Biologil Prouts n Vines (VACSERA, Ciro, Egypt). After n limtiztion perio of one week, the nimls were lssifie into four groups of equl verge oy weight n house in wireottome ges in room uner stnr onition of illumintion with 12 hours light-rk yle t 25+1 C. They were provie with wter n roent how liitum. All nimls reeive re in ompline with the Egyptin rules of niml experiments whih were pprove y the ethil ommittee of meil reserh of the Ntionl Reserh Centre, Ciro, Egypt Inution of Dietes Rts were fste for h efore inution ietes y using streptozotoin (STZ) from Sigm. St. Louis, MO, USA. (CAS no ). The rts reeive single intr-peritonel (i. p) injetion of 45 mg/kg/wt of STZ, whih ws freshly prepre n issolve in 0.05 M itrte uffer, ph 4.5 s esrie previously (Wei et l., 2003). Bloo gluose ws ontoure every 2 ys using n Auhek loo gluose meter (Rohe Dignostis, Bsel, Switzerln). Rts with loo gluose levels > 15 Mm (200mg/l) for 7 onseutive ys were onsiere ieti Experimentl Design Rts were ivie into four groups: group1, norml ontrol group in whih the rts reeive norml sline solution; group 2, ieti ontrol group in whih the rts were ieti using Streptozotoin (45mg/Kg.wt) n left untrete; group 3, group trete with Metformin in whih the ieti rts were trete with stnr orl hypoglyemi gent, metformin from Merk, CAS no , USA (100 mg/kg.wt) (Kosegw et l., 1999); n group 4, selenium nnoprtiles (SeNPs)-trete group, in whih the rts were trete with nnoselenium stilize in liposome (0.1 mg/kg. wt) orlly (Loeshner et l., 2014) in ition to metformin tretment. After 14 ys of ily tretment, overnight fsting nimls euthnize uner mil ether nesthesi Biosynthesis of Selenium Nnoprtiles Nnoselenium ws prepre oring to the metho of Dwivei et l.(2011) y simple wet hemil metho. Soium selenosulphte (N2SeSO3) (Sigm. St. Louis, MO, USA) preursor rete with ifferent orgni roxyli is in queous meium y using polyvinyl lohol to stilize selenium nnoprtiles. Then, the synthesize nnoprtiles were seprte from their solution y entrifugtion t 15,000 rpm n reisperse in queous meium with sonitor Chrteriztion of Selenium Nnoprtiles Chrteriztion of selenium nnoprtiles ws rrie out y exmintion of smple uner Trnsmission Eletron Mirosope (TEM) (JEOL-100 CX) Liposome Preprtion Elementl selenium nnoprtiles in molr rtio7:2 were use to prepre neutrl mutilmellr vesiles using the metho of Kim et l. (1985). Briefly, 10 mg of high purity L-lph-iplmitoyl phosphtiyl holine (DPPC) from (Lipoi KG-Germny) n 1 mg of selenium were trnsferre to 50 ml roun ottom flsk. Then, 15 ml of Chloroform ws e, n the flsk ws shken until ll lipis issolve in Chloroform. The solvent evporte uner vuum using rotry evportor until thin ry film of lipis ws forme. The flsk left uner vuum for 12 h to ensure the evportion of ll tres of hloroform. 10 ml of uffer (10 mm Trizm, ph 7) e to the flsk in mehnilly shken for 1 h t temperture of 45 C. The suspension entrifuge t 8000 rpm for 20 min n the superntnt isre. The liposome re-suspene in 10 ml uffer solution. Control liposome ws prepre following the sme lssil metho s efore using only

3 2017 Jorn Journl of Biologil Sienes. All rights reserve - Volume 10, Numer liquots of 10 mg of DPPC. The onentrtion of free selenium/ml of uffer ws juste to e 1/mg/ml DNA Frgmenttion Assy DNA frgmenttion ws mesure y spetrophotometer using iphenylmine (DPA) metho, oring to the metho of (Pernones et l., 1993) with some moifitions. Smples of liver were homogenize in lyses uffer ontining 5mM Tris HCl, ph 8.0, 20mM EDTA n 0.5% Triton X-100. Then, entrifuge t 1500 g for 20 min. Pellets were resuspene in 0.5N perhlori i n 5.5N perhlori i ws e to superntnt, entrifuge gin t 1500 g for 10 min to remove proteins. Smples were hete t 90 C n fter ool rete with iphenylmine (DPA) for 16 20h t room temperture. Asorne ws mesure t 600 nm using UV-oule em spetrophotometer (Shimzu 160 A). DNA frgmenttion in smples = (frgmente DNA in superntnt.)/[(frgmente DNA in superntnt intt + DNA in pellet)] were expresse s perentge of totl DNA ppering in the superntnt frtion Detetion of Oxitive DNA Dmge (Comet Assy) Aoring to the metho of Singh et l. (1988), 0.5 g of rushe smples were trnsferre to 1 ml ie-ol PBS. This suspension ws stirre for 5 min n filtere. Cell suspension (100 μl) ws mixe with 600 μl of low-melting grose (0.8% in PBS). 100 μl of this mixture ws spre on pre-ote slies. The ote slies were immerse in lyses uffer (0.045 M TBE, ph 8.4, ontining 2.5% SDS) for 15 min. The slies were ple in eletrophoresis hmer ontining the sme TBE uffer, ut evoi of SDS. The eletrophoresis onitions were 2 V/m for 2 min n 100 ma. Stining ws one using 20μg/ml ethiium romie t 4 C. A totl of 100 rnomly pture omets from eh slie ws exmine t 400 x mgnifition using fluoresene mirosope onnete to CCD mer to n imge nlysis system [komet 5 imge nlysis softwre evelope y Kineti Imging, Lt. (Liverpool, UK)]. A omputerize imge nlysis system quires imges, omputes the integrte intensity profiles for eh ell, estimtes the omet ell omponents n, then, evlutes the rnge of erive prmeters. To quntify the DNA mge, Til Length (TL), the perentge of migrte DNA (Til DNA %) n Til Moment (TM) were evlute. Til length (length of DNA migrtion) is relte iretly to the DNA frgment size n presente in mirometers. It ws lulte from the entre of the ell. Finlly, the progrm lultes til moment Bone Mrrow Chromosoml Aerrtion Assy At first, the mie were injete with 4 mg/kg.wt. olhiine two hours efore srifie. Metphse ells were prepre oring to the stnr tehnique of Preston et l. (1987). Bone mrrow ells were spirte from oth femurs of eh niml; then, the ells were entrifuge t 1000 rpm for 10 min. n resuspene in pre-wrme (37 C), hypotoni solution (0.075M potssium hlorie) for 20 min t 37 C. The smples were entrifuge n fixe in ol 3:1 methnol: glil eti i. Eh smple ws wshe five times fixtive. The slies were stine in 10% uffere Giems (ph 7.0), irrie n mounte in DPX. Chromosome errtions were ientifie oring to riteri esrie y Svge (1975) The Mironuleus Test Bone mrrow slies were prepre oring to the metho esrie y Hyshi et l. (1983). The one mrrow ws wshe with 1 ml of fetl lf serum n then smere on len slies. The slies were left to ir-ry n then fixe in methnol for 5 minutes followe y stining in My-Grunwl- Gemis for 5 minutes then wshe in istille wter n mounte. For eh niml, t lest 2000 polyhromti erythroytes (PCEs) per niml were exmine for the presene of mironulei Sttistil Anlysis Sttistil nlysis ws rrie out with SPSS softwre. Dt were nlyze using one-wy nlysis of vrine (ANOVA) followe y Dunn's post ho test for omprison etween ifferent tretments. The vlues were expresse s men ± S.E n ifferenes were onsiere s signifint when < P Results 3.1. Chromosomes Exmintion The nlysis of one mrrow hromosomes in ieti nimls epite vrious types of hromosoml errtions, whih inlue gp, rek, eletions, frgments, entromeri ttenutions n enomitosis s struturl errtions s well s polyploiy n preiploiy s numeril errtions. The results in tle 1 represent the hromosoml errtions nlysis n showe tht the frequenies of struturl n numeril hromosoml errtions were signifintly inrese (P< 0.05) in ieti nimls (STZhllenge group) ompring to ontrol group. In ontrst, the ieti nimls trete with Metformin h signifint (P< 0.05) ereses in the frequenies of ll struturl n numeril hromosoml errtions thn ieti group. In ition, metformin/nnoselenium group h the lowest frequenies of most struturl errtions (gp, frgment, enomitosis, C.A, totl struturl errtions) n totl numeril hromosome errtions reltive to Metformin group.

4 P P Jorn Journl of Biologil Sienes. All rights reserve - Volume 10, Numer 3 Tle 1. Effet of metformin n nnoselenium tretments on the frequeny of hromosome errtions in STZ- inue ieti rts Tretment Control D Gp 0.67±0.21P 3.67±0.21P Brek 0.33±0.21P 2.67±0.21P Struturl errtions Deletion Frgment Centromeri ttenution Enomitosis 0.33±0.21P 2.67±0.33P 0.17±0.16P 2.50±0.22P 0.50±0.22P 4.50±0.22P 0.67±0.21P 3.67±0.21P Totl 2.17±0.40P 18.33±0.42P Preiiploiy 1.33±0.21P 4.67±0.21P Numeril errtions Polyploiy 0.33±0.21P 2.50±0.22P Totl 1.67±0.21P 7.17±0.30P Totl errtions 3.84±0.30P 25.50±0.56P D+M 2.00±0.26P 1.50±0.22P 1.33±0.21P 1.50±0.22P 2.33±0.21P 2.00±0.26P 9.17±0.47P 1.50±0.22P 0.67±0.21P 2.17±0.16P 11.34±0.42P D+M+Se 1.17±0.30P 1.46±0.19P 1.17±0.23P 0.50±0.22P 0.83±0.16P 1.17±0.30P 4.50±0.34P 1.17±0.16P 0.33±0.21P 1.33±0.21P 5.83±4.77P D: Dieti rts y using streptozotoin (STZ); D+M: Dieti group trete with metformin rug; D+M+Se: Dieti group trete with metformin n nnoselenium prtiles. All t re expresse s men ± SEM. Mens ering ifferent letters supersripts re signifintly ifferent t (P < 0.05) Influene of Dietes on the DNA Dmge (Comet Assy) The DNA mge resulte from ietes ws investigte y omet ssy. the lengths of the omets (DNA tils) epene on the effet of ieti in rts, longer tils initing more DNA mge. Figure 1 shows representtive exmples of ieti rts. Control liver rt ells showe no tils (Figure A). Tils ppere in ieti rts (Figure B) n were sustntilly less in ieti tretment one (Figures C n D). The men vlue of til length in Dieti group ws inrese rpily n signifintly more thn tht of their ontrols, s shown in tle 2, n more DNA mge ws oserve in the til when ompre with ontrol group (Figure 1). As shown in tle 2, the extent of DNA mge, mesure in TM, inrese rpily in ieti group when ompre to ontrol. Trete groups with metformin or metformin+ nnoselenium showe signifint erese in omet TM vlues when ompre to ieti group (P<0.05) ut is still signifint inrese ompre with the ontrol. The mximum extent of DNA mge, inrese rpily in ieti group when ompre to ontrol. Then, the extent of mge erese signifintly (P<0.05) in oth trete groups with metformin or metformin+nnoslenium ut it i not reh the ontrol level. Tle 2. Comet prmeters in liver ells of metformin, nnoselenium tretments in STZ- inue ieti rts s mesure y omet ssy. Tretment Control D D+M D+M+Se Tile % 2.33± 0.33P 21.0± D: Dieti rts y using streptozotoin (STZ); D+M: Dieti group trete with metformin rug; D+M+Se: Dieti group trete with metformin n nnoselenium prtiles. All t re expresse s men ± SEM. Mens ering ifferent letters supersripts re signifintly ifferent t (P < 0.05) Mironuleus Test 0.58P 17.67± 0.33P 15.0± 0.58P Untile % 97.67± 0.33P 79.0± 0.58P 82.33± 0.33P 85.0± 0.58P Til length µm 2.16± 0.09P 5.27± DNA % 1.33± 0.22P 6.57± Til moment (unit) 2.84± 0.41P 34.65± The effet of ietes n tretments with metformin n nnoselenium on mironulete polyhromti erythroytes (MnPCEs) formtion in the one mrrow ells of mle rts is shown in Figure 2. In this Figure, it is emonstrte tht there were few formtions of MN in rts elong to ontrol group (3.33±0.21), ut MN formtion signifintly inrese (P<0.05) in ieti nimls (26.00±0.25) ompring to ontrol group. Menwhile, trete groups with metformin signifintly reue the MNPCE when ompre with ieti group. Aitionlly, MNPCE in the omine metformin n nnoselenium (14.00±0.25) signifintly reue more thn the group trete with metformin lone (20.67±0.33). 0.18P 4.5± 0.29P 3.77± 0.28P 0.28P 4.27± 0.15P 3.47± 0.26P 2.41P 19.25± 1.66P 13.19± 1.88P Figure 1. Effet of metformin, nnoselenium tretments in STZinue ieti rts on one mrrow DNA s etete y omet ssy. (A) Norml untrete ell; (B) ieti ell, () ieti rt trete with metformin n (D) ieti rt trete with nnoselenium n metformin. Figure 2. Frequenies of mironulete polyhromti erythroytes (MPCEs) in rt one mrrow ells in ll experimentl groups.

5 0T 2017 Jorn Journl of Biologil Sienes. All rights reserve - Volume 10, Numer DNA Frgmenttion The effets of nnoselenium n metformin rug on ieti rts were investigte y DNA frgmenttion ssy (Figure 3). The result emonstrte tht frgmente DNA, in the ontrol group (8.33±0.33), ws lower thn tht in the other trete groups. Also, frgmente DNA in ieti nimls (24.33±0.33) ws signifintly higher thn tht of the ontrol group (P< 0.05). On the other hn, trete nimls with metformin erese the DNA frgmenttion ompring with ieti nimls (P<0.05). In ition, the omine tretment with metformin /nnoselenium (12.17±0.47) erese the frgmenttion of DNA ompre to the tretment with metformin lone (18.33±0.33) (P<0.05). Figure 3. Perentges of DNA frgmenttion in rt liver ells in ll experimentl groups. 4. Disussion The present stuy inite tht the ministrtion of STZ inrese geneti ltertions (mironulei frequeny, frgmente DNA, s well s hromosoml errtions). The genotoxi effets of STZ, oserve in the present stuy, re in greement with the results otine from previous stuies (Vikrm et l., 2007; Atti et l., 2009).0T Also, 1TMrtínez-Pérez0T et l. (2007)0T foun tht there were high levels of mironuleus frequeny in type 2 ietes with no mirovsulr or mrovsulr omplitions. In ition, Cori et l. (2014) inite tht there is n ssoition etween T2DM n DNA Dmge y stuying the frequeny of mironulete polyhromti erythroytes. This genotoxiity my e ue to the presene of hyperglyemi onition in T2DM tht use proution of retive oxygen speies (ROS) (Phm-Huy et l., 2008; Roertson, 2004). Aumultion of ROS use n oxitive stress, whih plys ruil role in ellulr mehnism of tissue injury in wie spetrum of isese sttes. It my le to onstnt thret to ll living orgnisms n ntioxint efense system is employe y the oy to eliminte it (Ceoni et l., 2003). In ition, stress uses inrese resistne of insulin n isturne in pnreti β-ell funtion (Houstis et l., 2006). Oxitive stress n ttk ll types of mromoleules inluing DNA y ttking the ell memrne, nuleus n then geneti mterils, whih les to hromosoml errtions (Atti, 2010; Otton et l., 2004). DNA oxitive mging ours ue to moifitions in nuleotie ses or sugrs, these moifitions le to muttions whih my use formtion of tumors if they our in somti ells (Selvkumr et l., 2006), then erly ging my our whih les to eth (Rehmn et l., 1999). Moreover, ROS mge the ell y ifferent pthwys, suh s vne formtion of vne glytion en proution, hexosmine pthwy, protein kinse n lipi peroxition (Pioni et l., 2003), whih is result of ttking ROS to the resiues of polyunsturte ftty i of ell memrne. The imlne etween proution of ROS n the ility of ntioxint to etoxify their effet use oxitive stress (Ehty, 2007; Roerts n Sinhu, 2009). By using omet ssy, the present stuy revele tht ietes use DNA mge. This result is similr to tht otine from the stuy of Srs et l. (2001) who reporte inrese in oxitive DNA mge in type 1 n type 2 ietes, n these mge ws higher in type 2 ietes ompre to type 1 ietes. Also, Mrr et l. (2002) foun tht oxitive stress is inrese in ietes n this les to riovsulr isese. Dieti nimls trete with Metformin showe erese of most frequenies of struturl n numeril hromosome errtions n reue the iniene of MN formtion in PCEs in the ieti nimls. These my e ue to the lowering of loo gluose y Metformin s result of stimulting insulin relesing from funtioning pnreti et ells (Chunying et l., 2006; Kirpihnikov et l., 2002) n hepti gluose output s well s enhning peripherl gluose uptke (Biley n Turner, 1996), these to erese fsting loo gluose y 20 to 4%, erese oy weight, erese low ensity lipoprotein n inrese high ensity lipoprotein (Howlett n Biley, 1999). Therefore, ietes ptient hs three times norml rte of gluoneogenesis ut metformin reues this y one-thir (Hunl et l., 2000). Metformin inrese the insulin sensitivity n reue the oxitive stress levels n the tivity of tlse n superoxie ismutse when ompre with ietes rts (Vilel et l., 2016). So, the ul therpy with metformin rug promotes more enefits to oxitive stress ontrol in rts. The urrent t revele tht the tretment with SeNPs in ition to metformin rug resulte in signifint reution in the geneti ltertions (mironuleus formtion, hromosoml errtions) s well omet ssy. This my e ue to the ntioxint property of ompoun whih oul ply signifint role in eresing the nuler injury use y ietes (Aler et l., 2009). These oservtions re onsistent with those of Aelleem et l. (2016) who foun tht selenium nnoprtiles possesses ntioxint n nti-ieti tivities y eresing oxitive stress iomrkers s well s loo gluose level. Similrly, Ahme et l. (2016) foun tht selenium nnoprtiles hve ntiieti poteny, repression of oxitive stress, potentiting of the ntioxint efense system, n inhiition of pnreti inflmmtion. So, selenium plys protetive role ginst type 2 ietes (Steinrenner n Sies, 2009). The protetive roles of selenium in mmmlin ell re ue to its funtion in the tive site of mny ntioxint enzymes, suh s thioreoxin reutse, glutthione n GR (Flor

6 Jorn Journl of Biologil Sienes. All rights reserve - Volume 10, Numer 3 et l., 2002). Dietry selenium my use n inrese in glutthione peroxise, whih hs the ility to etoxify ROS (Köhrle n Gärtner, 2009) tht n interfere with insulin signling n result in regultion of gluose levels n prevention of ietes (Golstein et l., 2005). Selenium is lso si omponent of selenoprotein, n importnt enzyme in the oy whih protets from oxitive stress n inflmmtion (Rymn, 2012). The nutritionl efiieny of selenium uses musulr ystrophy, enemi ftl riomyopthy, n hroni egenertive iseses when use lone or in omintion (Rymn, 2002). 5. Conlusion In onlusion, the present stuy emonstrte the ntiieti potentil of SeNPs elivere in liposomes in the experimentl moel of T2DM. This effet ppere in eresing the frequenies of hromosoml errtions, in ition to eresing the numer of MNPCEs s well s repiring mge DNA whih is represent in DNA frgmenttion ssy n omet ssy. Thus, SeNPs my enefit in linil purposes, espeilly in enhning the effet of T2DM. Referenes Aelleem RMA, elhmee HF, Askr ME, Hssn SHM n Eltl AI Moultory role of selenium nnoprtiles n grpe see extrt mixture on oxitive stress iomrkers in ieti irrite rts. Inin J Phrmeutil Eu Res., 50 (1): Aler AI, Shw EJ, Stokes T n Ruiz F Newer gents for loo gluose ontrol in type 2 ietes: summry of NICE guine. BM J. 338: 1668 Ahme HH, A El-Mksou MD, Ael Moneim A.E n Agln HA Pre-Clinil stuy for the ntiieti potentil of selenium nnoprtiles. Biol Tre Elem Res., 177(2): Atti SM Deleterious effets of retive metolites. Oxitive Me Cellulr Longevity, 3 (4): , Atti SM, Hell GK n Alhier A A Assessment of genomi instility in norml n ieti rts trete with metformin. Chemio-Biologil Intertions, 180 (2): Biley CJ n Turner RC Metformin. N Engl J Me., 334:579. Ceoni C, Borso A, Crgoni A n Ferrri R Oxitive stress in riovsulr isese. Arh. Biohem. Biophys., 420: Chunying C, Hongwei Y, Jiujing Z, Bili LQ, Shuiping L, Peiqun Z n Zhifng C The roles of serum Se n selenoproteins on Hg toxiity in environmentl n ouptionl exposure. Environ. Helth Perspet.,114 (2): Cori SCT, Bstos AS, Orrio SRP, Seolin R, Rquel A. Sntos D, Tkhshi CS n Srel-Cming RM Elevte mironuleus frequeny in ptients with type 2 ietes, yslipiemi n Perioontitis. Mutgenesis, 29 (6): Dwivei C, Shh CP, Singh K, Kumr M n Bjj PN An orgni i-inue synthesis n hrteriztion of selenium nnoprtiles. J Nnotehnol., 2011: 1 6. Ehty KS Mitohonril unoupling proteins-wht is their physiologil role? Free Ril Biol Me., 43 (10): Frrokhin A, Bhmni F, Tghizeh M, Mirhshemi SM, Ari MH, Rygn F, Aghvo E n Asemi Z Selenium supplementtion ffets insulin resistne n serum hsrp in ptients with type 2 ietes n oronry hert isese. Horm Met Res., 48 (4): Flor SJS, Knnn GM, Pnt BP n Jiswl DK Comine ministrtion of oxli i, suimer n its nlogue for the reversl of gllium rsenie inue oxitive stress in rts. Arh Toxiol., 76: Go X, Zhng J n Zhng L Hollow sphere selenium nnoprtiles: their in-vitro nti hyroxyl ril effet. Av Mter., 14: Gtes B, Myers B, Cttle B n Xi Y Synthesis n hrteriztion of uniform nnowires of trigonl selenium. Av Funt Mter., 12: Golstein BJ, Mhev K n Wu X Reox prox: insulin tion is filitte y insulin-stimulte retive oxygen speies with multiple potentil signling trgets. Dietes 54: Hyshi M, Sofuni T n Ishite M Jr An pplition of riine ornge fluoresent stining to the mironuleus test. Mutt Res., 120: Houstis N, Rosen ED n Lner ES Retive oxygen speies hve usl role in multiple forms of insulin resistne. Nture, 440: Howlett HC n Biley CJ A risk-enefit ssessment of metformin in type 2 ietes mellitus. Drug Sf., 20 (6): 489. Hunl R, Krssk M, Dufour S, Lurent D, Leon V, Chnrmouli V, Inzuhi S, Shumnn W, Petersen K, Lnu B n Shulmn G Mehnism y whih metformin reues gluose proution in type 2 ietes. Dietes, 49 (12): IDF. Interntionl Dietes Feertion. Retrieve 29 Novemer Kim S, Jos RE n White SH Preprtion of multilmellr vesiles of efine size-istriution y solventspherule evportion. Biohim Biophys At., 812 (3): Kirpihnikov D, MFrlne SI n Sowers JR Metformin: n upte. Ann Intern Me., 137 (1): Kithi AE, Umpierrez GE n Miles JM Hyperglyemi rises in ult ptients with ietes. Dietes Cre, 32 (7): Köhrle J n Gärtner R Selenium n thyroi. Best Prt Res Clin Enorinol Met., 23 (6): Kosegw I, Chen S, Awt T, Negishi K n Ktym S Troglitzone n metformin, ut not glienlmie, erese loo pressure in Otsuk Long Evns Tokushim ftty rts. Clin Exp Hypertens., 21(3): Liu W, Li X, Wong YS, et l Selenium nnoprtiles s rrier of 5-fluorouril to hieve ntiner synergism. ACS Nno., 6: Loeshner K, Hrup N, Hnsen M, Pereir SA, Gmmelgr B, Møller LH, Mortensen A, Lm HR n Lrsen EH Asorption, istriution, metolism n exretion of selenium following orl ministrtion of elementl selenium nnoprtiles or selenite in rts. Metllomis, 6(2): Mkhluf SBD, Au-Mukh R, Ruinstein S, Breitrt H n Genken A Moifie PVA-Fe3O4 nnoprtiles s protein rriers into sperm ells. Smll, 4:

7 2017 Jorn Journl of Biologil Sienes. All rights reserve - Volume 10, Numer Mrr G, Cotroneo P, Pitoo D, Mnto A, Di Leo MA, Ruotolo V, Cputo S, Girin B, Ghirln G n Sntini SA Erly inrese of oxitive stress n reue ntioxint efenses in ptients with unomplite type 1 ietes: se for gener ifferene. Dietes Cre, 25: Mrtínez-Pérez LM, Cer-Flores RM, Gllegos- Criles EC, Dávil-Roríguez MI, Irr-Costill E n Cortés- Gutiérrez EI Frequeny of mironulei in Mexins with type 2 ietes mellitus. Prgue Me. Rep., 108: Mruthur NM, Tseng E, Hutfless S, Wilson LM, Surez-Cuervo C, Berger Z, Chu Y, Iyoh E, Segl JB n Bolen S Dietes meitions s monotherpy or metformin-se omintion therpy for type 2 ietes: A Systemti Review n Met-nlysis. Annls of Internl Meiine 7 june. Mueller AS, Mueller K, Wolf NM n Plluf J Selenium n ietes: n enigm? Free Ri Res., 43(11): Nvrro-Alro n M n Lo pez-mrtı nez MC Essentility of selenium in the humn oy: reltionship with ifferent iseses. Si Totl Environ., 249: Niset JC, Sturtevnt JM n Prins JB Metformin n serious verse effets. Me J Aust., 180: Olnsky L, Mrhetti A n Lu H Multienter retrospetive ssessment of thizoliineione monotherpy n omintion therpy in ptients with type 2 ietes: omprtive sugroup nlyses of glyemi ontrol n loo lipi levels. Clin Ther. 25: Otton R, Sorino F.G, Verlengi R n Curi R Dietes inues poptosis in lymphoytes. J. Enorinol., 182: Pernones CE, Iller VA, Pekhm D, Stunz LL n Ashmn R F Regultion of poptosis in vitro in mture murine spleen T ells. J Immunol.,151: Phm-Huy LA, He H n Phm-Huy C Free rils, ntioxintsin isese n helth. Inter J Biomeil Si., 4 (2): Pioni, L, Qugliro L n Ceriello A Oxitive stress in ietes. Clin Chem L Me., 41: Preston RJ, Den BD, Gllowy S, Holen H., MFee AF n Shelly M Mmmlin in vivo ytogeneti ssys: Anlysis of hromosome errtions in one mrrow ells. Mutt. Res., 189: Ro L, M Y, Zhung MJ, Luo TJ, Wng YY n Hong A Chitosn-eorte selenium nnoprtiles s protein rriers to improve the in vivo hlf-life of the peptie therpeuti BAY for type 2 ietes mellitus. Inter J Nnomeiine, 9: Rymn M The rgument for inresing selenium intke. Pro Nutr So., 61: Rymn MP Selenium n humn helth. Lnet, 379: Rehmn A, Nourooz-zeh J, Moller W, Tritshler H, Pereir P n Hlliwell B Inrese oxitive mge to ll DNA ses in ptients with type 2 ietes mellitus. FEBS Lett., 448: Roerts CK n Sinhu KK Oxitive stress n metoli synrome. Life Si., 84 (21): Roertson R Chroni oxitive stress s entrl mehnism for gluose toxiity in pnreti islet et ells in ietes. J Biol Chem., 279: Srs S, Yilmz M, Oztok U, Cki N n Krky AE Assessment of DNA strns rekge y omet ssy in ieti ptients n the role of ntioxint supplementtion. Mutt Res., 490: Svge JRK Clssifition n reltionship of inue hromosoml struturl hnges. J Me Genet., 12: Selvkumr E, Prhlthn C, Vrlkshmi P et l., Moifition of ylophosphmie-inue lstogenesis n poptosis in rts y lph-lipoi i. Mutt Res., 606: Shi Y n Frnk BHU The glol implitions of ietes n ner. The Lnet, 383(9933): Singh NP, MCoy MT, Tie RR n Shneier EL A simple tehnique for quntittion of low levels of DNA mge in iniviul ells. Exp Cell Res., 175: Steinrenner H n Sies H Protetion ginst retive oxygen speies y selenoproteins. Biohim Biophys At., 1790: The Amerin Soiety of Helth- System Phrmists. "Metformin Hyrohlorie. Retrieve Jn Vikrm DN, Tripthi P n Jen Rmro G B Evlution of streptozotoin genotoxiity in rts from ifferent ges using the mironuleus ssy. Regultory Toxiol Phrmol., 49: Vilel DD, Peixoto LG, Teixeir RR, Bptist NB, Cixet DC, e Souz AV, Mho HL, Pereir MN, Silv RS n Espinol FS The role of metformin in ontrolling oxitive stress in musle of ieti rts. Oxitive Me Cellulr Longevity, Artile ID , 9 pges. Vos T, Flxmn AD, Nghvi M, Lozno R, Mihu C, Ezzti M, Shiuy K, Wng H, Zhng J n Yu H Elementl selenium t nno size possesses lower toxiity without ompromising the funmentl effet on selenoenzymes: omprison with selenomethionine in mie. Free Ri Biol Me., 42: Wng XL, Yng TB, Wei J, Lei GH n Zeng C Assoition etween serum selenium level n type 2 ietes mellitus: non-liner ose-response met-nlysis of oservtionl stuies. Nutr J., 15 (1): 48. Wei M, Ong L, Smith MT, Ross FB, Shmi K, Hoey AJ, Burstow D n Brown L The streptozotoin-ieti rt s moel of the hroni omplitions of humn ietes. Hert Lung Cir., 12 (1): Worl Helth Orgniztion. "Aout ietes. Retrieve 4 April Worl Helth Orgniztion. "Dietes Ft sheet N 312". Otoer Zhng J, Wng H, Yn X n Zhng L Comprison of short-term toxiity etween Nno-Se n selenite in mie. Life Si., 76:

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