Available online at International Journal of Current Research Vol. 9, Issue, 10, pp , October, 2017

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1 z Aville online t Interntionl Journl of Current Reserch Vol. 9, Issue, 1, pp , Octoer, 217 INTERNATIONAL JOURNAL OF CURRENT RESEARCH ISSN: X RESEARCH ARTICLE COMPARATIVE STUDY ON THE INFLUENCE OF DIFFERENT DRUGS AND NUTRIENTS AGAINST ALUMINIUM- INDUCED D NEPHROTOXICITY AND HEPATOTOXICITY IN RATS 1, *Azz A. i, 1 Toq M. Elnhhs, 2 Aeer I. Ad El-Ftth, 1 Mon M. Kml nd 1Krem Au-Elfotuh 1Deprtment of Phrmcology nd Toxicology, Fculty of Phrmcy, -Azhr University, Ciro, Egypt 2Deprtment of Biochemistry, Fculty of Phrmcy, -Azhr University, Ciro, Egypt ARTICLE INFO ABSTRACT Article History: Bckground: Environmentl pollution with the different luminium () contining compounds especilly Received 22 nd July, 217 those in industril wste wter exposes people to higher thn norml levels of tht represents n Received in revised form environmentl nmentl risk fctor. Cosmetics, wre nd continers re lso sources of esides some foods nd 13 th August, 217 food dditives. In ddition to its known neurotoxicity, ffects other ody structures like skeletl system, Accepted 24 th Septemer, 217 lood cells, liver nd kidney. Accumultion of in kidney nd liver induces nephrotoxicity nd Pulished online 17 th Octoer, 217 heptotoxicity. toxicity. Coenzyme Q1 (CoQ1) is pseudo -vitmin sustnce primrily present in the mitochondri. ondri. It is powerful ntioxidnt nd cts s rdicl scvenger. Whet grss is nturl product tht contins crohydrtes, proteins, vitmins, minerls, enzymes nd hs ntioxidnt, nti-inflmmtory, Key words: nticncer ncer nd crdiovsculr protection ctivities. Coco is n excellent source of iron, potent ntioxidnts uminum, nd cn protect ginst mny diseses. Vinpocetine is n ntioxidnt nd nti inflmmtory while zinc is n Nephrotoxicity, essentil trce element involved in cell division nd its deficiency is oserved in mny types of liver disese. Heptotoxicity, Ojective: To evlute nd compre the potency of different drugs nd nutrients (CoQ1, whetgrss, Coenzyme Q1, coco, vinpocetine nd zinc) ginst nephro- nd hepto-toxicity induced y in rts. Whetgrss, Methods: Rts were divided to seven groups nd received dily for three weeks either sline for control Coco, group or Cl 3 (7 mg/kg, IP) for -toxicity model groups. Five groups of -toxicity model (treted Vinpocetine, groups) were orlly received together with ech of the following; CoQ1 (2mg/kg), whet grss Zinc. (1mg/kg), coco powder (24mg/kg), vinpocetine (2mg/kg) or zinc (32mg/kg). Biochemicl chnges in the serum levels of nine minotrnsferse (ALT), sprtte minotrnsferse (AST), lkline phosphtse htse (ALP), lctte dehydrogense (LDH) s well s totl iliruin, cholesterol, triglycerides, glucose, cretinine nd ure were mesured. Liver nd kidney specimens from ll groups were lso collected for the ssessment of heptic nd nephrotic level of inflmmtory meditors (TNF-α, IL-6β, nucler fctor kpp B (NF -κb), Cspse-3, oxidtive prmeters (MDA, SOD, TAC, NO) nd DNA frgmenttion. Histopthologicl chnges in liver nd kidney were lso evluted. Results: s: Three weeks of Cl 3 (7 mg/kg, IP) exposure induced nephro- nd hepto-toxicity in rts. Tretment y the ll used drugs showed protection ginst hzrds of Cl 3. The protective effects were indicted y the significnt decrese in ALT, AST, ALP, LDH s well s totl iliruin, cholesterol, *Corresponding uthor: triglycerides, glucose, cretinine nd ure levels which were incresed y. Liver nd kidney of the treted Azz A. i, groups showed decrese in MDA, NO, TNF-α, IL-6β, NF-κB, cspse-3 nd DNA frgmenttion which Deprtment of Phrmcology nd were incresed y, together with significnt increse in SOD nd TAC which were decresed y. The Toxicology, Fculty of Phrmcy, protection tion ginst oth nephro- nd hepto-toxicity ws more pronounced especilly with CoQ1 nd whet grss thn the other used drugs. Histopthologicl exmintions confirmed the iochemicl results of -Azhr University, Ciro, Egypt. toxicity nd of protection. Conclusion: Protection from nephrotoxicity, heptotoxicity nd the consequent degenertions induced y cn e chieved y using different drugs s CoQ1, whetgrss, coco, vinpocetine nd zinc, ut CoQ1 s well l s whet grss possesses the most superior protection. Copyright 217, Azz A. i et l. This is n open ccess rticle distriuted under the Cretive Commons Attriution License, which permits unrestricted use, distriution, nd reproduction in ny medium, provided the originl work is properly cited. Cittion: Azz A. i, Toq M. Elnhhs, Aeer I. Ad El-Ftth, Mon M. Kml nd Krem Au-Elfotuh, 217. Comprtive Study on the Influence of Different Drugs ginst uminium- induced Nephrotoxicity nd Heptotoxicity in Rts, Interntionl Journl of Current Reserch, 9, (1), INTRODUCTION uminum () is regrded s the third most undnt element nd the most common metl in the erth s crust (Frin et l., 22). Exposure to is principlly through drinking wter, food (spices, corn nd yellow cheese), phrmceuticl compounds (ntcids, deodornts, vccines nd llergen injections), utensils nd the environment (Yokel et l., 28, nd Exley, 211). uminum is considered n environmentl nd industril pollutnt tht cuses rod spectrum of toxicity. The kidney is the min orgn for excretion (Exley

2 58685 Azz A. i et l. Comprtive study on the influence of different drugs nd nutrients ginst luminium- induced nephrotoxicity nd heptotoxicity in rts et l., 1996). Erly reports reveled tht ws not toxic with regrd to kidney function (Triq et l., 1999). Lter reports demonstrted tht plsm iochemicl ltertions, renl trophy nd morphologicl chnges of the Bowmn s cpsule nd severl different renl tuules resulted from toxicity (Belïd-Nouir et l., 213). Impirment of renl function occurred s consequence of these pthologicl chnges in the kidney structure, s disturnce in the norml nti-oxidtive system ws induced through the formtion of rective oxygen species (ROS) (Cmpell et l., 24). Lipid peroxidtion in liver s well s in kidney ws induced y toxicity (El- Demerdsh, 24 nd Kneko et l., 24). Chinoy & Ptel (1999) nd Moumen et l. (21) reported tht heptotoxicity nd chnges in the oxidtive sttus hve lso een reported fter exposure. Additionlly, Neill et l. (1996) nd Wilhelm et l. (1996) reported heptic ccumultion immeditely fter either n intrvenous or orl dministrtion oth in experimentl nimls nd humn eings. As inds to DNA, RNA, nd inhiits hexokinse, lkline phosphtses (ALPs) nd phospho-oxidse ctivities (Ochmnski & Brsz, 2). Coenzyme Q1 (CoQ1) is known to e n electron crrier in the electron trnsport chin. CoQ1 synthesized in the ody cells from tyrosine mino cid in the presence of dequte levels of vitmins such s folic cid (VrnesiÄ-Bender, 21). It hs een reported tht CoQ1 hve ntioxidnt nd ntipoptotic ctivities y regenertion of other ntioxidnts, so it hs een used s nti-ging nd is effective in the tretment of cognitive disorders (Spindler et l., 29 nd Dumont et l., 211). Vinpocetine is known to e n inhiitor of cyclic GMP phosphodiesterse, powerful legend of peripherl enzodizepine inding sites nd lso cn ct s locker of NV1.8 sodium chnnel ctivity (Ahn et l. 1989, Zhou et l. 23 nd Gulyás et l. 25). Besides, vinpocetine hs n ntiinflmmtory ction tht cn inhiit tumor necrosis fctorlph (TNF -α), inducing nucler fctor-kpp B (NF -κ B) ctivtion nd the induction of pro-inflmmtory meditors (Jeon et l. 21). Zinc is n essentil trce element which required in numer of iologicl ctions nd is nontoxic t physiologicl doses (Betholf, 1988). Severl reports hve indicted the vlule ctions of zinc under the conditions of oxidtive dmge (Cgen &Klssen, 1979, Cre et l., 1995, Goel et l., 27 nd Rishi et l., 28). Zinc stilizes the cell memrne structure through its ntioxidnt effects, which my e s result of its ility to regulte the levels of metllothioneins (Kng, 1999). On the other hnd, whetgrss (Triticum estivum L.) is known to e rodly used helth food, consumed commonly s fresh juice or s cpsules, tlets nd liquid concentrtes. Formultions of whetgrss possess different phrmcologicl effects such s ntioxidnt (Flcioni et l., 22), tumor suppressor (Ary nd Kumr, 211), hypoglycemic (Mohn et l., 213) nd neuro-protective effects (Jng et l., 21). Whet grss is considered n potent source of ntioxidnt enzymes, ntioxidnt vitmins such s A, B, C, E nd minerls like potssium, sulfur, zinc, clcium, colt, iron, phosphorus (Leoncini et l., 212 nd Stevenson et l., 212). Coco is known s one of the richest flvonoidcontining foods ville, it contins ntioxidnt polyphenols clled flvonols tht my exert hepto protective effects (Amin et l., 24, Serfini, 24 nd Cordero-Herrer et l., 215) nd nti mlril effects (Addi, 21 nd Amponsh et l., 212). It is fortuntely tht the ntioxidnt ctivities of coco unchnged fter different mnufcturing processes (Sthl et l., 29 nd Mleyki & Ismil, 21). Since, kidney nd liver re of the trget tissues of luminum toxicity s they involved in its elimintion or in the metolism nd detoxifiction (Exley et l., 1996 nd Ogueche et l., 214). So, the essence of the present study ws to evlute the iochemicl nd histopthologicl ltertion in the kidney nd the liver of rts exposed to. Additionlly, to compre the potency of different drugs nd nutrients s CoQ1, vinpocetine, zinc, whetgrss nd coco in the protection ginst nephrotoxicity nd heptotoxicity induced y. MATERIALS AND METHODS Animls Seventy mle Sprgue Dwley rts were used. Rts weighing g were otined from Nile Co. for Phrmceuticls nd Chemicl Industries, Ciro, Egypt. They were housed in stinless-steel cges (four per cge) under the sme dequte conditions, with lterntively 12 hour light nd drk cycles, t temperture of 25 ± 1 C. Rts were kept under the sme dequte conditions nd provided with their dily dietry requirements of stndrd diet pellets (El -Nsr, Au Zl, Ciro, Egypt) contined not less thn 2% protein, 5% fier, 3.5% ft, 6.5% sh nd vitmin mixture, wter ws given dliitum. Rts were tken to test sitution one hour efore ech experiment for dpttion nd fter removing food nd wter from the cges. The study ws conducted in the period from Mrch to July, 216 in ccordnce with ethicl guidelines of -Azhr University (Fculty of Phrmcy), Egypt. Drugs nd chemicls From Sigm Chemicl Co. (St. Louis, MO, USA); CoQ1 nd uminum chloride - hydrted (Cl 3.6H2O) were purchsed. Cl 3 ws freshly dissolved in distilled wter while CoQ1 ws suspended in 1% queous solution of Tween 8; suspensions were freshly prepred every dy. l other solvents nd chemicls were of the highest grdecommercilly ville. Vinpocetine ws suspended in 1% queous solution of Tween 8; suspensions were freshly prepred every dy. Whetgrss ws freshly dissolved in distilled wter s well s coc nd zinc sulphte. Experimentl design Methods Rts were rndomly ssigned to seven groups nd received dily for three weeks either sline for control group or injected (I.P) with 7 mg/kg Cl 3.6H2O ( i et l., 215) for luminum toxicity model groups. Five groups of -toxicity model (treted groups) were orlly received together with ech of the following; 2mg/kg of CoQ1 (Andressen et l., 1999), 1mg/kg of whet grss (Veer et l., 214), 24mg/kg of coco powder (Rozn et l., 27), 2mg/kg of vinpocetine (Rlf 7 Josef, 1991) or 32mg/kg of zinc sulphte (Agnieszk et l., 216). A-Biochemicl Investigtions Blood smpling At the end of the three weeks, lood smples were collected vi eye puncture from ech rt efore scrifiction into serum seprtor tues, llowed to stnd (3 min), centrifuged (3

3 58686 Interntionl Journl of Current Reserch, Vol. 9, Issue, 1, pp , Octoer, 217 rpm for 15 min), serum collected nd stored t -2 C until the ssy of the studied iochemicl prmeters. I- Blood iomrkers i-estimtion of renl functions Blood ure nitrogen (BUN) nd serum cretinine were mesured using quntittive colorimetric ure determintion (QuntiChrom ure ssy kit) (Biossy Systems, Hywrd, CA, USA) nd quntittive colorimetric cretinine determintion (QuntiChrom cretinine ssy kit).l procedures were performed ccording to the mnufcturers instructions. ii-estimtion of heptic functions Heptic function iomrker serum levels nmely; Serum lnine minotrnsferse (ALT), serum sp rtte minotrnsferse (AST), nd totl iliruin were estimted y colorimetric ssy kits (Biomed -dignostics, Ciro, Egypt), ccording to the methods descried y Tietz (1976) nd Mlloy nd Evelyn (1937). so, lkline phosphtse (ALP) ws estimted using k-ssy ELISA kit (Kmiy Biomedicl, Settle, WA, USA). Stnio Lortory Kits (Boerne, TX, USA) were utilized for the determintion of the serum lumin levels. l procedures were performed ccording to the mnufcturers instructions. Estimtion of LDH: LDH ws determined ccording to the method of Tietz (1976) (Biosystems S.A, Brcelon, Spin). iii-estimtion of cholesterol nd triglycerides Colorimetric ssy kits for the mesurement of serum cholesterol (totl nd HDL - cholesterol) nd triglycerides (Biomed-dignostics, Ciro, Egypt), were used in this study. iv-estimtion of glucose levels Stnio Lortory Kits (Boerne, TX, USA) were utilized for the determintion of the serum glucose levels. II-Tissue iomrkers At the end of the three weeks, rts were scrificed y decpittion then livers nd kidneys were dissected nd wshed with ice-cold sline. Liver nd kidney tissues were kept frozen t -8 C till the time of nlysis. They were homogenized in sline then the homogentes were used to ssess the oxidtive stress mrkers; s superoxide dismutse (SOD), totl ntioxidnt cpcity (TAC), lipid peroxides which were expressed s mlondildehyde (MDA) s well s Nitric oxide (NO) ws lso determined. Anti -inflmmtory mrkers were ssessed; y mesuring the levels of tumor necrosis fctor-α (TNF -α), interleukin -6 β (IL -6β) nd nturl fctor kpp- β (NFκ -B).Assessment of poptotic mrker; y mesuring Cspse-3 ctivity. Finlly, DNA frgmenttion ws done s well s histopthologicl exmintions; y tking specimens from ll kidney nd liver res from different groups. i-renl nd heptic oxidtive stress estimtion In the kidney nd liver homogentes, MDA nd SOD s well s TAC were mesured. Lipid peroxidtion cn e determined s MDA, y estimting the level of thiorituric cid rective sustnces (TBARS), ccording to the method of Stoh, (1978) using (Biodign ostic, Ciro, Egypt). SOD content ws ssessed, relying on the ility of the enzyme to inhiit the phenzine methosulphte medited reduction of nitrolue tetrzolium dye (Nishikimi et l., 1972), where the increse in sornce t 56 nm for 5 min ws mesured. On the other hnd, determintion of TAC is performed through the rection with defined mount of exogenously provide H 2 O 2. The residul H 2 O 2 is colorimetriclly determined y the enzymtic rection tht involves the conversion of 3, 5-dichloro-2- hydroxyenzene sulphonte to colored product (Korcevic et l., 21). For NO estimtion, vndium trichloride ws used to reduce nitrte to nitrite (Mirnd et l., 21). The method of nitrite estimtion is sed on Griess rection tht ws performed using the kit provided y Biodignostic (Ciro, Egypt). l procedures were performed ccording to the mnufcturers instructions. ii-assessment of inflmmtory mrkers. The involvement of inflmmtion ws ssessed y mesuring the levels of (TNF -α, IL-6β nd NFκ-B) in kidney nd liver homogente of ll groups, utilizing the commercilly ville rt Quntikine Rt TNF-α ELISA Kits (R&D Systems, MN, USA), RyBio Rt IL-6β (RyBiotech, Inc., USA) nd rt NFκ-B ELISA kit Cusio Biotech (Cusio Life Science, Inc., Chin) respectively. iii-assessment of poptotic mrkers Cspse-3 ctivity ws detected in the kidney nd liver homogentes using ELISA kit (MyBioSource Sn Diego, Cliforni, USA). The mnufcturer's instructions were followed precisely nd the developed color ws mesured spectrophotometriclly t 45 nm immeditely. iv-assessment of DNA frgmenttion. DNA frgmenttion% ssy ws conducted using the procedure supplied y Qigen kit (Hilden, Germny).To detect DNA frgmenttion. The DNA in the gel ws visulized nd photogrphed under UV light(r). In DNA lddering ssy, low moleculr weight frgments of DNA re extrcted selectively from the cells wheres the higher moleculr weight DNA stys ssocited with the nuclei. The isolted DNA is seprted y electrophoresis nd visulized using ethidium romide. DNA ws electrophoresed using 2% grose gel nd visulized y ultrviolet light following ethidium romide stining. B-Histopthologicl exmintion of the kidney nd liver. In 1% formlin for 24 h, kidney nd liver specimens were fixed then they were wshed with tp wter, they were prepred nd stined for light microscopy (Bncroft nd Stevens, 1996). For dehydrtion; seril dilutions of lcohol were used (methyl, ethyl nd solute ethyl). In hot ir oven t 56ºC for 24 h, specimens were clered in xylene emedded in prffin. By using microtome t 4 microns thickness, prffin ees wx tissue locks were sectioned. Then, sections were collected on glss slides nd deprffinized. They were stined for routine histologicl exmintion using Hemtoxylin nd Eosin stin for routine histopthologicl exmintion.

4 58687 Azz A. i et l. Comprtive study on the influence of different drugs nd nutrients ginst luminium- induced nephrotoxicity nd heptotoxicity in rts Sttisticl Anlysis Dt re presented s men ± SEM. Multiple comprisons were performed using one-wy ANOVA followed y Tukey Krmer s post hoc test. Unpired t-test ws used to compre two different tretments. The.5 level of proility ws used s the criterion for significnce. l sttisticl nlyses were performed using Instt (version 3) softwre pckge. Grphs were sketched using GrphPd Prism (ISI, USA) softwre (version 5). RESULTS Serum Enzymes Activities Cl 3 induced out two-fold mrked increse of liver enzymes ALT nd AST ctivities in the lood, compred with control Fig.1 & Fig.2 respectively. Administrtion of zinc, Q1, vinpocetine, coco, nd whet grss, ws found effective to reduce tht increse significntly y (58%, 38%, 61%, 41% nd 44% respectively) of ALT enzyme ctivity, nd y (52%, 35%, 54%, 44% nd 56% respectively) of AST enzyme ctivity s compred to Cl 3 group. With regrd to ALP enzyme ctivity, 76% significnt increse ws oserved with Cl 3 tretment, while (24%, 13%, 33%, 16% nd 14%) mrked decrese noticed in zinc, Q1, vinpocetine, coco, nd whet grss treted groups respectively Fig.3. 29% nd 16%) ws presented fter tretment with zinc, Q1, vinpocetine, coco nd whet grss respectively. On the other hnd, (13%, 28%, 17%, 31% nd 25%) significnt increse in the HDL level with zinc, Q1, vinpocetine, coco nd whet grss tretment respectively s compred with intoxicted group. AST (u/ml) Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 2. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on serum AST ctivity in rts + Vinpocetine + Coco +Whet grss ALT (u/ml) 4 ALP (u/ml) Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 1. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on serum ALT ctivity in rts Serum Totl Biliruin + Vinpocetine + Coco + Whet grss As shown in Fig. 4 Cl 3 incresed totl iliruin level significntly y 36%, ut zinc, Q1, vinpocetine, coco, nd whet grss supplementtion meliorted it registering significnt decrese of (54%, 46%, 85%, 53% nd 54% respectively). Results reveled (3% nd 34%) significnt increse in cholesterol, nd triglyceride levels respectively in the Cl 3 treted group Fig.5 & 6. A significnt decrese (34%) in HDL level ws presented in Fig.7 s compred with control group. Co-dministrtion with the protective drugs zinc, Q1, coco, or whet grss meliorted the previous hyperlipedemic chnges significntly with Cl 3 treted group (7%, 15%, 19% nd 18% respectively) except vinpocetine, which hd no significnt chnge in cholesterol level. Similrly, the significnt reduction in triglyceride level (2%, 18%, 41%, + Vinpocetine AL+Coco + Whet grss Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 3. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on serum ALP ctivity in rts Totl ilir uin(mg/m l) Vinpocetine + Coco + Whet grss Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 4. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on serum Totl iliruin level in rts

5 58688 Interntionl Journl of Current Reserch, Vol. 9, Issue, 1, pp , Octoer, 217 Serum Lipid Profile Cholesterol (mg/ml) Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 5. Effect of oth lone, nd in comintion with Zn, CoQ1, Vinpocetine, Coco, or Whet grss on serum Cholesterol level in rt TG (mg/ml) Vinpocetine + Coc + whet grss Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 6. Effect of oth lone, nd in comintion with Zn, CoQ1, Vinpocetine, Coco, or Whet grss on serum Triglycerides level in rts 8 5 A + Vinpocetine + Coco + Whet grss Glu cose ( mg/ml) Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 8. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco, or Whet grss on lood Glucose level in rts Glucose Level in Blood Fig.8 reveled (36%) significnt increse of Cl 3 treted group compred with control. Zinc co-dministrtion, incresed glucose level significntly (14%).While coco, decresed it significntly (8%). Neither significnt difference ws found with vinpocetine or with whet grss s compred to Cl 3 -intoxicted group. umin level 5 umin level decresed significntly in Cl 3 intoxicted group y 39%. Tretment with zinc, Q1, vinpocetine, coco, nd whet grss succeeded to increse it y (21%, 3%, 27%, 3% nd 3% respectively) Fig.9. Renl Toxicity Blood Mrkers + Vinpocetine Cl 3 (7 mg/kg, IP) for three weeks in rts, induced significnt increse in the lood mrkers of renl toxicity BUN, nd cretinine concentrtions (hlf - fold, nd 3 -folds respectively) compred with control. The BUN concentrtion in Fig.1 ws significntly decresed in rts treted with zinc, Q1, vinpocetine, coco nd whet grss (17%, 36.5%, 3.6%, 28% nd 36% respectively) s compred to Cl 3 -intoxicted group. Likewise, the serum cretinine level in Fig.11 ws significntly decresed in rts treted with zinc, Q1, vinpocetine, coco nd whet grss (3%, 25%, 22%, 38% nd 24% respectively) s compred to Cl 3 toxicted group. + Coc + whet grss HDL (m g/m l) umin ( g/dl) C ontrol A l A l + Q1 A l + Vinpoc etine + C oc o + Whet grss Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 7. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco, or Whet grss on serum HDL-C level in rts + Vinpocetine + Coco + Whet grss Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 9. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on serum umin level in rts

6 58689 Azz A. i et l. Comprtive study on the influence of different drugs nd nutrients ginst luminium- induced nephrotoxicity nd heptotoxicity in rts BUN (mg/ml) 2 + Vinpocetine + Coco + Whet grss (A) Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p < Fig. 1. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco, or Whet grss on BUN level in rts C re tin in e (mg /ml).5. Co ntrol + Vinpocetine + Coco + W het grss Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 11. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco, or Whet grss on serum Cretinine level in rts Inflmmtory meditors (Tissue specimens, kidney & Liver) TNF-α level s shown in Fig.12A, 12B presented out one nd hlf-fold significnt increse in oth kidney nd liver tissues fter tretment with Cl 3 compred to control., The decrese ws significntly in kidney tissue y (12%, 2%, 53%, 17%, nd 9%) of Zinc, Q1, vinpocetine, coco, nd whet grss tretments respectively. No significnt difference with Q1 in liver tissue, while zinc, vinpocetine, coco, nd whet grss tretments decresed the cytokine level y (2%, 28%, 25%, nd 16%respectively) of liver tissue, compred to Cl 3 treted group. IL-6β level incresed reltively one nd hlf- fold in kidney tissue, nd incresed mrkedly two nd hlf- fold in liver tissue, fter tretment withcl 3 compred to control s shown in Fig.13A, 13B. Heptic TNF (P g/ m l) A l + Zn + Vinpocetine A l + Coco + Whet grss (B) Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5 : Significnt difference from - treted group t p <.5. Fig. 12. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on renl nd heptic TNF-α level (pg/mg protein) in rts The inflmmtory meditor level decresed significntly in oth kidney tissues y (41%, 36%, 47%, 55% nd 44%), nd liver tissues y (36%, 26%, 41%, 32% nd 28%) with the codministrtion of zinc, Q1, vinpocetine, coco nd whet grss respectively s compred to Cl 3 treted group. NF- k B level incresed one nd hlf-fold in kidney tissues, while incresed to gret extent in liver tissues five-fold, versus fter Cl 3 tretment compred to control Fig.14A,14B the elevted nucler fctor level reduced mrkedly y the drugs zinc, Q1, vinpocetine, coco, nd whet grss dministrtion(15%, 15%, 19%, 23%, nd 15%) in kidney tissues, nd (43%, 62%, 4%, 6% nd 29%) in liver tissues respectively s compred to Cl 3 treted group. Cspse-3 poptotic meditor level ws enhnced triple-fold significntly in kidney, ut quietly significnt increse one nd hlf-fold in liver homogente, under the effect of Cl 3 tretment compred to control Fig.15A, 15B ll the drug tretments zinc, Q1, vinpocetine, coco, nd whet grss hd eneficil effect on the elevted meditor y (57%, 29%, 48%, 45% nd 37%) decrese in kidney tissues. However, only zinc, Q1 nd vinpocetine, succeeded to reduce the elevted level significntly y (2%, 12% nd 24% respectively) in liver tissues s compred to Cl 3 treted group.

7 5869 Interntionl Journl of Current Reserch, Vol. 9, Issue, 1, pp , Octoer, Renl IL-6B (Pg/ m l) 4 2 Heptic IL-6B (pg/ml) 1 5 Co ntrol (B) : Significnt difference from the control group t p <.5, : Significnt difference from - (A) Vlues were expressed s men ± SEM. treted group t p <.5. + Vinp ocetine + Coco + Whet grss Fig. 13. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on renl nd heptic IL-6β level (pg/mg)in rts + Vinpocetine + Coco + Whet grss Renl NF-kB (reltive expression) Heptic NF-kB (reltive express (A) : Significnt difference from the control group t p <.5, : Significnt difference from - Vlues were expressed s men ± SEM. treted group t p <.5. + Vinpocetine + Coco + Whet grss Fig. 14. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on renl nd heptic NF-kB level in rts (B) npocetine + Coco het grss Renl Cspse ( ng/mg) Heptic cspse (ng/mg) Vinpocetine + Coco + Whet grss + Vinpocetine + Coco + Whet grss (B) : Significnt difference from the control group t p <.5, : Significnt difference from - (A) Vlues were expressed s men ± SEM. treted group t p <.5. Fig. 15. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on renl nd heptic Cspse-3 level in rts

8 58691 Azz A. i et l. Comprtive study on the influence of different drugs nd nutrients ginst luminium- induced nephrotoxicity nd heptotoxicity in rts Renl MDA (mmol /mg protein) Heptic M DA (mmol/mg protein) Vinpocetine + Coco + Whet grss (A) (B) Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 16. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on renl nd heptic MDA level in rts. + Vinpocetine + Coco + Whet grss 3 4 Renl SOD(u/mg) 2 1 Heptic SOD (u/mg) Vinpocetine + Coco + Whet grss Cont rol + Vinpocetine + Coco + Whet grss (A) (B) Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 17. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on renl nd heptic SOD ctivity in rts Renl TAC ( nmol/mg ) 2 1 Heptic TAC (nmol/ mg ) 1 + Vinpocetine + Coco + Whet grss + Vinpocetine + Coco + Whet grss (A) (B) Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 18. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on renl nd heptic TAC level in rts

9 58692 Interntionl Journl of Current Reserch, Vol. 9, Issue, 1, pp , Octoer, Renl NO (nmol/mg) 4 2 Heptic NO (nmol/mg) AL + Vinpocetine + Coco + Whet grss + Vinpocetine + Coco + Whet grss (A) (B) Vlues were expressed s men ± SEM. : Significnt difference from the control group t p <.5, : Significnt difference from - treted group t p <.5. Fig. 19. Effect of oth lone nd in comintion with Zn, CoQ1, Vinpocetine, Coco or Whet grss on renl nd heptic NO level in rts Antioxidnt prmeters kidney & liver (Tissue specimens) MDA level in Cl 3 treted group ws incresed significntly y out two-folds in kidney tissue homogente nd threefolds in liver tissue homogente Fig.16A, 16B s compred with control. Tretment with zinc, Q1, vinpocetine, coco nd whet grss decresed it significntly y (16%, 7%, 24%, 17% nd 11%respectively) of kidney tissue nd y (32%, 39%, 37%, 14% nd 37%respectively) of liver tissue compred to Cl 3 treted group. SOD ctivity mrkedly reduced 65% in kidney tissue nd 55% in liver tissue Fig. 17A, 17B y the influence of Cl 3 intoxiction, compred with control. Reversed results otined fter tretment with zinc, Q1, vinpocetine, coco, nd whet grss y significnt elevtion to (39%, 141%, 15%, 129%, nd 68% respectively) of kidney tissue, nd to (46%, 54%, 54%, 69% nd 38%respectively) of liver tissue compred to Cl 3 treted group. TAC ws decresed significntly in oth kidney nd liver tissue homogente 37%, nd 44% respectively upon exposure tocl 3, compred to control Fig. 18A, 18B Zinc, Q1, vinpocetine, coco nd whet grss tretment cme ck to within norml rnge toto (36%, 59%, 29%, 18% nd 53%respectively) of kidney tissue nd (38%, 39%, 47%, 41% nd 27% respectively) of liver tissue compred to Cl 3 treted group. NO produced gretly out six-folds in kidney tissue nd four-fold increse in liver tissue s result of Cl 3 tretment s compred to control Fig. 19A, 19B. DNA Frgmenttion 1- Kidney frgment Fig.2 A Lne D: shows DNA streks (Groups of AL) (Induced toxicity which found in the model (M) lddering shpe) Group P: there re no streks groups of tretment 2- Liver frgment Fig.2 B An grose gel electrophoresis show DNA frgmenttion Lne M: DNA mrker with 1p Group C: there re no streks Lne D: shows DNA streks (Groups of AL) (Induced toxicity which found in the model (M) lddering shpe) Group P: there re no streks groups of tretment On the other hnd, Zinc, Q1, vinpocetine, coco, nd whet grss tretments were le to reduce the elevted level y (68%, 41%, 66%, 63% nd 51%respectively) of kidney tissue, nd y (52%, 75%, 42%, 73% nd 6% respectively) of liver tissue compred to Cl 3 treted group. Histopthologicl chnges of kidney & liver Norml histologicl structure ppered of the structure of the glomeruli nd tuules t the cortexnd lso norml histologicl of centrl vein nd surrounding heptocytes in the prenchym control rts group Fig.21. An grose gel electrophoresis show DNA frgmenttion Lne M: DNA mrker with 1p Group C: there re no streks : Kidney tissue A: liver tissue Fig. 21. Histopthologicl chnges in rts tissues of control group

10 58693 Azz A. i et l. Comprtive study on the influence of different drugs nd nutrients ginst luminium- induced nephrotoxicity nd heptotoxicity in rts : Kidney tissue C: liver tissue Fig. 23. Effect of nd Zinc on histopthologicl chnges of rts tissues d: Kidney tissue D: liver tissue B: liver tissue Fig. 22. Effect of on histopthologicl chnges of rts tissues Fig.22 showed oedem with firosis. Detected in the thick renl cpsule (1), degenertion in the lining tuulr epithelium (2), Hyperplsi & dysplsi were noticed in the lining epithelium of some focl tuules (3) in the treted groups. While Fig.22B Showed thickening in the heptic cpsule.by inflmmtory cells infiltrtion nd firolstic cells prolifertion (B1) etween the heptocytes in the heptic prenchym (B2, B3). Inflmmtory cells infiltrtion (B4) in the portl re, diffusekupffer cells, prolifertion etween the heptocytes (B5). Fig.23c showed renl tuules showed degenertion. Chng nd cogultive necrosis (c1,c2), focl re of the tuules showed hyperplsi nd dysplsi in the lining epithelium (c3), focl hemorrhge nd inflmmtory cells infiltrtion in cortico-medullry junction (c4,5) of treted group plus zinc dministrtion. While Fig.23C Showed Cpsule ws thick y firous Connective tissue prolifertion (C1), firosis with inflmmtory cells infiltrtion etween the heptocytes in the prenchym (C2) & Ftty chnges in heptocytes. Fig. 24. Effect of nd COQ1 on histopthologicl chnges of rts tissues No histopthologicl ltertion in kidney tissues in group treted with COQ1 s shown in Fig.24d. While Fig.24D Showed thickening in the heptic cpsule y oedem nd inflmmtory cells infiltrtion. Thickening in the renl cpsule y oedem nd inflmmtory cells infiltrtion re shown in Fig.25e of treted group dministered with vinpocetine. Fig.25E Showed thickening of heptic cpsule, y oedem nd inflmmtory cells infiltrtion, heptocytes showed poptosis (E1), inflmmtory cells infiltrtion nd firosis were detected in the portl re (E2). In Fig.26f kidney tissues showed, thickening in the cpsule y inflmmtory cells infiltrtion nd firolstic cells prolifertion. Fig.26F Showed inflmmtory cells infiltrtion nd firolstic cells prolifertion in the thick cpsule (F1), collgen fiers extended etween heptocytes in the prenchym (F2), Apoptosis in some individul heptocytes surrounding the centrl vein (F3, F4), congestion in the portl res, periductl inflmmtory cells infiltrtion nd oedem (F5). e: Kidney tissue c: Kidney tissue

11 58694 Interntionl Journl of Current Reserch, Vol. 9, Issue, 1, pp , Octoer, 217 E: liver tissue Fig. 25. Effect of nd vinpocetine on histopthologicl chnges of rts tissues f: Kidney tissue F: liver tissue Fig. 26. Effect of nd coco on histopthologicl chnges of rts tissues Kidney tissues in Fig.27 g Showed tuules degenertion nd cogultive necrosis (g1,g2). While Fig.27 G Showed thickening in the heptic cpsule y oedem nd inflmmtory cells infiltrtion (G1) extended in the prenchym etween the heptocytes (G2). Congestion in the portl vein (G3) ws present. DISCUSSION uminum ccumultion is common in ll tissues of mmmls, such s the kidneys, liver, lood, ones, hert, nd rin (- Khtni, 21) nd it ws reported tht the toxic ctions of were prominent in the kidney s well s the liver (Aukr et l., 24 nd Jinyu et l., 216). In the present study, injection of rtes y Cl3 (7 mg/kg, IP) dily for consecutive three weeks induced significnt increse in lood ure nitrogen (BUN) nd serum cretinine concentrtions in the Cl3-intoxicted group when compred to control. These findings were in ccordnce with El-Demerdsh, (24) reported tht exposure induces chnges in kidney function. Recently, Jinyu et l. (216) lso reported tht BUN ltertions indicte tht suppression of glomerulr filtrtion function occurred s result of Cl3 exposure. On the other hnd, the current study reveled significnt increse in the serum levels of ALT, AST, ALP, LDH, totl iliruin nd significnt decrese in serum lumin in the Cl3-intoxicted group when compred to control. As mtter of fct, the increse in trnsminses levels re encountered in conditions inducing heptocellulr dmge, cell memrne functionl integrity loss nd necrosis (Ninh et l., 23). Cell necrosis induces n increse in LDH enzyme concentrtions in serum nd tissue. The LDH relesed into the medium considered n index of cell deth nd memrne permeility to LDH s result of cell memrne disintegrtion nd enzyme lekge (Shen et l., 1995 nd Lindell et l., 1996). Gonzlez et l. (27) nd Tripthi et l. (28) demonstrted lso tht interctions etween oxidtive stress nd heptic dmge my enhnce the progression of chronic hepto-degenertive diseses. Additionlly, the present work reveled significnt elevtion in serum lipid profile (mnifested y significnt increse in serum TG nd TC with significnt decrese in HDL-C) in Cl 3 -intoxicted group compred to the control group. This cme in ccordnce with other studies reported tht triggered dyslipidemi my led to vriety of heptic normlities. Li et l. (26) nd Milloux et l. (27) reported n ccumultion of lipids in the liver in chronic kidney disese resulting from overlod. There is lnce etween the oxidnts (rective oxygen species [ROS]) nd ntioxidnts in helthy individuls. If this lnce is ltered s result of over production of ROS, oxidtive stress my occur, which ffects oxidtive dmge to orgns (Joshi et l., 213). In prllel to these results, the oxidtive mrkers of the current study in renl nd heptic tissues, showed significnt elevtion in renl nd heptic MDA nd NO while significnt decline in ntioxidnt ctivity of renl nd heptic SOD nd TAC were lso reported. These results re in ccordnce with Ferretti et l. (23), Cndn & Tuzmen (28) nd Milloux et l. (211), demonstrted tht ltered redox sttus nd rised lipid peroxidtion re considered hllmrks of n oxidtive environment, nd these dysfunctions re ll linked to the toxicity. Therefore, the suggested mechnism y which cn inflict its toxic effects is; creting free rdicls in the ody. It cuses toxic ction due to its ility to trnsfer electrons which cn ffect cell integrity, producing lipid peroxidtion in the intrcellulr memrnes, ffecting lso its permeility of sucellulr orgnelles, the structure nd functions of proteins nd nucleic cids (Tus et l., 213). Additionlly, Aukr et l. (23) scertined tht even minute quntities of luminum in heptocytes ssocited with ROS increse nd peroxidtion. Newiry et l. (29) supported this suggestion lso s n increse in the level of thiorituric cid rective sustnce (TBARS) nd decline in the ctivities of GST, SOD nd CAT in liver, kidney nd rin of rts treted with (34 mg/kg ody weight Cl3 dily for 7 dys) were reported. Grrel et l. (1994) nd Bondy et l. (1998) hve reported tht luminum-enhnced peroxidtion my e relted to luminum-induced nitric oxide synthse (NOS) ctivity nd rised NO products in rt rin tissue nd microglil cells. NO is generted from L-rginine y the id of NOS enzyme which formed in vriety of tissues nd included in vrious physiologicl nd pthologicl processes (Moncd et l.,

12 58695 Azz A. i et l. Comprtive study on the influence of different drugs nd nutrients ginst luminium- induced nephrotoxicity nd heptotoxicity in rts 1991). Wrd et l. (21) suggest nother mechnism for toxicity, s exposure could enhnce disruptions in the minerl lnce, leding to ions replcing iron nd mgnesium, which would then result in decline in Fe +2 inding to ferritin. Hence, -induced free iron ions relesing from iologicl complexes cn ctlyze hydroperoxides decomposition to hydroxyl rdicls through Fenton s rection. The initition of lipid peroxidtion, cusing memrne dmge occurred s result of this high hydroxyl rdicl rectivity. Regrding the reltionship etween Cl3 nd immune function in rts, the current results exhiited significnt elevtions in the levels of pro-inflmmtory cytokines; including TNF-α nd IL-6β nd NFκ-B in kidney nd liver tissues of Cl3- intoxicted group, which fits with the results of Mnn et l. (213) who mentioned tht chronic inflmmtory process might contriute to Cl3 toxicity. Kuo et l. (211) illustrted the role of Nucler fctor kpp et (NF-κ B) in the inflmmtion s it controls the expression of different genes encoding pro-inflmmtory cytokines, nd inducile enzymes such s cyclooxygense-2 (COX -2) nd inducile nitric oxide synthse (inos) leding to NO nd TNF-α production. The elevted inflmmtory stte in our emerging dt confirmed with histopthologicl exmintion of the kidney nd liver sections of the Cl3 - intoxicted group. Firstly, the kidney homogente showed oedem with firosis nd thickening of renl cpsule, degenertion in the lining tuulr epithelium, hyperplsi nd dysplsi were noticed in the lining epithelium of some focl tuules.these findings were in greement with previous results of in vivo nd in vitro studies demonstrting tht Cl3 increses the thickness of Bowmn s cpsule sement memrne, resulted in trophy of glomerulus cpillries nd proximl nd distl tuules dmge (Stcchiotti et l., 26 nd Srgzi et l., 26). Secondly, the histopthologicl exmintion of liver tissues in the present study confirmed the previous results, s it reveled thickening in the heptic cpsule y inflmmtory cells infiltrtion nd firolstic cells prolifertion etween the heptocytes in the heptic prenchym. Besides, inflmmtory cells infiltrtions in the portl re, kupffer cells diffusion, prolifertion etween the heptocytes were lso reported.these findings were in ccordnce with Bogdnovic et l. (28) reported congestion of centrl vein, sinusoidl dilttion nd lipid ccumultion in liver. To explore the mechnism y which Cl3-induce poptosis, cspse-3 protein level ws mesured in kidney nd liver homogentes. The current work showed significnt elevtion in the level of renl nd heptic cspse 3 in Cl3- intoxicted group. These results potentited y oserving DNA frgmenttion in liver nd kidneyin Cl3- intoxicted group (Lne D: shows DNA streks, Fig. 2A & 2B). The hllmrk of poptosis is DNA frgmenttion (Ngt, 2). Zou et l. (1997) suggested the moleculr mechnisms underlying poptosis including cytochrome relese, cspses ctivtion, condenstion of chromtin, DNA frgmenttion s well s ded cells phgocytosis, nd deris y scvenger cells. Therefore, Cl 3 induced poptosis through the ctivtion of cspse-3, y the following consequences; rective oxygen species (ROS) increse the mitochondril memrne permeility leding to mitochondril filure (Hung et l. 28). The mitochondril memrne permeility is dependent upon the mitochondri permeility trnsition pore tht results in cytochrome c relese from the mitochondri nd into the cytosol (Yng et l. 214). Once relesed, cytochrome cinds A pf-1 in the cytoplsm forming complex tht cn ctivte cspse-9 with susequent ctivtion of deth-inducingcspse-3. (Ghrii et l. 22). Due to the previous nephrotoxic effects of injection of Cl 3 (7 mg/kg, IP) for successive three weeks, the present study ws designed to evlute nd compre the potency of different drugs nd nutrients(coq1, vinpocetine, zinc, whetgrss nd coco) ginst nephrotoxicity induced y in rts. The nephro-protective potency of the studied drugs nd nutrients ginst toxicity reveled significnt improvement in the kidney function ctivity in -treted groups with different drugs nd nutrients s CoQ1 (2mg/kg), whet grss (1mg/kg), coco powder (24mg/kg), vinpocetine (2mg/kg) or zinc (32mg/kg) when compred with -toxicity model. There ws significnt decline in the serum levels of BUN nd cretinine. Additionlly, CoQ1 treted group showed the more significnt decline in the serum level of BUN when compred to the other treted groups. These findings cme in greement with severl studies hve demonstrted tht some of the renl protective effect of CoQ1 in rts with renl dysfunction proly relted to its ntioxidnt effect (Mnning et l., 25 nd Liu et l., 26). CoQ1 is known to e memer of the mitochondril electron trnsport chin, which cn ccept either one or two electrons. It cts s powerful nturl ntioxidnt, oxygen-derived free rdicl scvenger nd s memrne stilizer (Ostrowski, 1999). Regrding the oxidtive stress mrkers in kidney homogente, the current study demonstrted significnt decline in renl MDA nd NO, while significnt increse in renl SOD nd TAC ws lso reported in -treted groups with different drugs nd nutrients when compred with -toxicity model. Interestingly, there ws predominnt significnt increse in level of renl SOD in CoQ1 treted group when compred to other treted ones. Additionlly, there ws predominnt significnt increse in level of renl TAC in CoQ1 followed y whet grss treted group when compred to other treted ones. These results re in prllel with McCrthy et l. (24) reported tht in niml models, CoQ1 hve protective ctivities ginst toxin-induced oxidtive stress. The recent study of Syed et l. (215) lso proposed tht CoQ1 might e vlule s potent cellulr defense ginst oxidtive dmge fter luminum toxicity. He elieved tht in the presence of induced cell toxicity, CoQ1 cn increse cytochrome c oxidse ctivity; consequently, should help to restore mitochondril ctivity nd ATP production. To interpret the present nephro-protective results of whet grss treted group, Khn et l. (213) suggested tht the ntioxidnt enzymes present in whet grss helps rid of free rdicls vi the regultion of cellulr homeostsis nd ugmenttion of self-defense to oxidtive stress. For the evlution of the ntiinflmmtory nd nti-poptotic properties of the studied drugs nd nutrients, the present work reveled significnt decrese in renl TNFα, IL-6β, NFκ-B nd cspse 3 in -treted groups with different drugs nd nutrients when compred with -toxicity model. These findings were confirmed y DNA frgmenttion in the kidney s, Lne D: shows DNA streks (Groups of AL-induced toxicity which found in the model (M) lddering shpe) Group P: there re no streks groups of tretment (Fig. 2A).

13 58696 Interntionl Journl of Current Reserch, Vol. 9, Issue, 1, pp , Octoer, 217 Additionlly, the current results reveled more significnt decrese in renl TNFα level in vinpocetine followed y CoQ1treted groups when compred with other treted groups. These results cme in ccordnce with Jeon et l. (21) who reported tht Vinpocetine exerts n ntiinflmmtory ction inhiiting (TNF -α)-induced nucler fctor-kpp B (NF -κb) ctivtion, nd the induction of proinflmmtory meditors. In ccordnce with these findings lso, Schmelzer et l. (27) suggests tht CoQ1 hs numer of independent nti-inflmmtory ctivities. The novelty of the present results of nephroprotective evlution tht the histopthologicl exmintion of kidney reveled tht the most nephroprotective drug ws CoQ1 s, no histopthologicl ltertion ws oserved which suggest tht CoQ1of prominent protection ginst the nephrotoxic effects of toxicity. In contrst with other drugs which showed thickening in the renl cpsule y oedem nd inflmmtory cells infiltrtion. These results cme in ccordnce with severl studies which suggest the nti-inflmmtory mechnism of CoQ1 y reduction of pro-inflmmtory cytokines secretion in monocytes nd lymphocytes fter n inflmmtory stimulus through n influence on the expression of NFκ-B -dependent genes(sohet et l., 29, Bentinger et l., 21 nd Mohseni et l., 214). Furthermore, in series of reports in ptients with coronry rtery disese, CoQ1 used s tretment(6 3 mg CoQ1/1dy for 12 weeks), to reduce oxidtive stress nd improve the ntioxidnt enzyme ctivity s well s lowering inflmmtion s ssessed y plsm levels of inflmmtory mrkers such s tumor necrosis fctor (TNF)-α nd interleukin (IL)-6 (Lee et l., 212, 212 nd 213). Similrly, dietry CoQ1 (diet supplemen ted with.7%.7% (w/w) CoQ1 for26 weeks) ws ccompnied with decline in plsm oxidtive stress nd inflmmtory mrkers in rt model of the metolic syndrome (Kunitomo et l., 28). There re lso, lot of niml studies which suggest tht ntioxidnt ctivities of CoQ1hve eneficil effects on kidney disese (Mnning et l., 25, Ngse et l., 26 nd Liu et l., 29). Secondly, the heptoprotective potency of the studied drugs nd nutrients ginst toxicity reveled significnt improvement in the liver function ctivity in -treted groups with different drugs nd nutrients [CoQ1 (2mg/kg), whet grss (1mg/kg), coco powder (24mg/kg), vinpocetine (2mg/kg) or zinc (32mg/kg)] when compred with -toxicity model. As there ws significnt decline in the serum levels of ALT, AST, ALP, LDH s well s totl iliruin nd significnt increse in heptic synthetic function in treted groups (mesured y serum lumin). These results were in prllel with other studies of Bhsin et l. (214) nd Lmuel-Rvent et l. (25) suggested tht zinc nd coco hve ntioxidnt ctivity, so e useful in preventing the toxic effects of on the liver. These results re in continution of erlier studies oserved remrkle improvement in the levels of trnsminses following zinc dministrtion (Dhwn et l., 1992; Dhwn nd Goel, 1994). The protective effect is relted to the role of zinc in the protein metolism regultion, which in turn regultes the levels of trns minses. The current study demonstrted significnt decline in heptic MDA nd NO, while significnt increse in ntioxidnt heptic SOD nd TAC ws lso reported in -treted groups with different drugs nd nutrients when compred with -toxicity model. In ccordnce with these results Adrmoye et l. (28) reported tht increses oxidtive stress vi incresing the level of superoxides (O2 ) nd peroxides (H2O2). Since these oxides were not mesured in this work, ut it is elieved tht ROS cn ind nd rect with heptic cellulr components inducing heptic injury, nd sodeteriorting liver function (Adekunle et l., 29). Tking these mechnisms into considertion. Adekunle et l. (29) lso stted tht drugs tht hve ntioxidnt effect or hve the ility to decrese oxidtive stress cn e of vlule role in inhiiting the d effects of on the liver. Similr oservtions hve lso een reported y Esprz et l. (25) in experimentl nimls treted with zinc fter exposure. This could e relted to the ntioxidnt chrcter of zinc, which ws le to normlize the ctivity of SOD. The present results lso showed the more prominent heptoprotective effect of CoQ1 (2mg/kg), to t oxicity model rther thn other treted groups, s significnt decrese in heptic MDA nd NO levels ws reported when compred to other treted groups. These results cme in ccordnce with Ostrowski, (1999) stted tht CoQ1 cts s potent nturl ntioxidnt, oxygen-derived free rdicl scvenger nd s memrne stilizer (Ostrowski, 1999). In ddition, CoQ1 exerts inhiiting chrcter on mitochondril ROS genertion nd inner mitochondril depolriztion (Kwong et l., 22). Moreover, CoQ1 supplementtion protects plsm memrne ginst oxidtive stress (Gómez- Díz et l., 23).The ove mentioned effects of CoQ1 permit this coenzyme to exhiit n improvement in ech of the oxidtive stress mrkers s investigted in the present study. The present work reveled significnt decrese in heptic TNFα, IL-6β, NFκ-B nd cspse 3 in -treted groups with different drugs nd nutrients when compred with -toxicity model. These findings were confirmed y DNA frgmenttion in the liver s, Lne D: shows DNA streks (Groups of ) (induced toxicity which found in the model (M) lddering shpe) while group P: there is no streks in the groups of tretment (Fig.2B). Additionlly, Co enzyme Q1 (2mg/kg), treted group showed the more significnt decline in the level of heptic NFκ-B when compred to the other treted groups. Besides, the histopthologicl exmintion of Co enzyme Q1 treted group showed lso slight thickening in the heptic cpsule y oedem nd inflmmtory cells infiltrtion when compred to toxicity model which cn indicte possile heptoprotective role of Co enzyme Q1 supplementtion ginst toxicity in rts. These results were in greement with severl studies demonstrted tht supplementtion of the diet with CoQ1 hs een found to ffect plsm ntioxidnt nd inflmmtory mrkers in helthy individuls(gökel et l., 21) nd niml studies hve demonstrted effects on liver oxidtive stress nd lipid metolism (Cno et l., 29, Sohet et l., 29, Sfwt et l., 29). In conclusion, this study showed protective effect of using different drugs nd nutrients s CoQ1, whetgrss, coco, vinpocetine nd zinc ginst Cl3 induced renl nd heptic dmge. Interestingly, CoQ1 s well s whet grss possess the most superior protection. It is our elief tht CoQ1 should e le to not only llevite oxidtive stress t the level of the mitochondri nd consequently increse cell survivl, ut is lso likely to help to resolve liver nd kidney dysfunction present in ptients with luminum toxicity. Reduction of oxidtive stress nd inflmmtion my e considered s the min pthwys of ction. However, further experiments t the moleculr levels re required to illustrte clerly the mechnism y which these drugs nd nutrients reverse Cl 3 induced nephrotoxicity nd heptotoxicity.