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			<p class="Type-of-Article">Research Article</p>
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			<p class="title- ParaOverride-1">The Protective Effect of  Pometone (Pomegranate Seed Oil) on Oxidants /Antioxidant Status in Methionine Overload Rabbits (Part III)</p>
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			<p class="Authors ParaOverride-1">&nbsp;</p>
			<p class="Authors ParaOverride-1"><span class="CharOverride-1">Baraa Najim Al-Okaily*, Lekaa Najim Abdulla, Khalisa Khadim Khudair</span></p>
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			<p class="Affiliations ParaOverride-1">Department of Physiology and Pharmacology, College of Veterinary Medicine, University of Baghdad-Iraq.</p>
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			<p class="Abstract ParaOverride-1">&nbsp;</p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-2">Abstract</span> |<span class="CharOverride-2"> </span>This study was aimed to investigate the role of pomegranate seed oil (PSO) as antioxidant in alleviating the deleterious effects of methionine overload in some biochemical aspects of adult female rabbits. Thirty two female rabbits were randomly assigned into four equal groups and given orally the following treatment for 42 days: the first groups was drenched corn oil, serving as control group (C), the second group (T<span class="CharOverride-3">1</span>) was intubated with methionine 100mg/kg. B.W, the third groups (T<span class="CharOverride-3">2</span>) was intubated with methionine 100mg/kg. B.W and pomegranate seed oil (PSO) 30 mg /Kg. B.W., while animals in the group T<span class="CharOverride-3">3</span> were intubated with pomegranate seed oil 30 mg/Kg. B.W.<span class="CharOverride-4"> </span>Fasting blood samples were collected at 0, 21 and 42 days of experiment to assess: serum total cholesterol(TC), malondialdehyde (MDA), peroxynitrite radicals, reduced glutathione (GSH) concentrations and lactate dehydrogenase activity( LDH). At the end of the current study group T<span class="CharOverride-3">1</span> showed a significant increase in TC, MDA, peroxynitrite radicals concentrations and LDH activity with significant reduction in GSH concentration. Besides the results showed a significant decrease in serum TC and peroxynitrite radical concentrations with significant elevation in GSH concentration in group T<span class="CharOverride-3">3</span> as compared with the control. It can be concluded that intubation of PSO correct the oxidative stress in methionine overload rats.</p>
		  <p class="Abstract ParaOverride-1">&nbsp; </p>
			<p class="Abstract ParaOverride-1"><span class="CharOverride-2">Keywords</span> | Pomegranate seed oil, Methionine, Oxidative stress, Antioxidants</p>
		  <p class="Abstract ParaOverride-1">&nbsp;</p>
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		<div class="Basic-Text-Frame">
			<p class="Editor----Citation"><span class="CharOverride-2">Editor</span> | Kuldeep Dhama, Indian Veterinary Research Institute, Uttar Pradesh, India.</p>
			<p class="Editor----Citation"><span class="CharOverride-2">Received</span> | March 06, 2015; <span class="CharOverride-2">Revised</span> | May 25, 2015; <span class="CharOverride-2">Accepted</span> | May 27, 2015; <span class="CharOverride-2">Published</span> | June 07, 2015&#9;&#9;</p>
			<p class="Editor----Citation"><span class="CharOverride-2">*Correspondence</span> | Baraa Najim Al-Okaily, University of Baghdad, Iraq; <span class="CharOverride-2">Email:</span> baraanajim@yahoo.com</p>
			<p class="Editor----Citation"><span class="CharOverride-2">Citation </span>| Al-Okaily BN, Abdulla LN, Khudair KK (2015). The protective effect of pometone (pomegranate seed oil) on oxidants /antioxidant status in methionine overload rabbits (part III). Adv. Anim. Vet. Sci. 3(7): 377-383.  </p>
			<p class="Editor----Citation"><span class="CharOverride-2">DOI</span> | <a href="http://dx.doi.org/10.14737/journal.aavs/2015/3.7.377.383"><span class="Hyperlink">http://dx.doi.org/10.14737/journal.aavs/2015/3.7.377.383</span></a></p>
			<p class="Editor----Citation"><span class="Editor---Citation CharOverride-2">ISSN (Online)</span> | 2307-8316; <span class="Editor---Citation CharOverride-2">ISSN (Print)</span> | 2309-3331</p>
			<p class="Editor----Citation"><span class="CharOverride-2">Copyright</span> © 2015 Al-Okaily et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</p>
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		<div class="Basic-Text-Frame">
			<p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Heading-1--Introduction----">INTRODUCTION</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Caps-on-First-Para"><span class="_idGenDropcap-1">M</span><span>ethionine </span>is an essential amino acid found in both animal and plant proteins. Methionine is a protective factor against various types of liver damage, but excessive dietary methionine is hepatotoxic (<a href="#Sukhanov-DS--Vinogradova-TI--Zabolotnykh-NV--Vasil-eva-SN--Vitovskaia-ML--2013-"><span class="Hyperlink">Sukhanov et al., 2013</span></a>). When the metabolic pathways of methionine are interrupted due to malnutrition or gene mutation, homocysteinemia (Hcy) level will be elevated (<a href="#Jakubowski-H--2006-"><span class="Hyperlink">Jakubowski, 2006</span></a>). New suggestion revealed that hyperhomocysteinemia (HHcy) play an important role in different physiological disorders including, lipid peroxidation (<a href="#Bouzouf-M--Martinez-Cruz-F--Molinero-P--Guerrero-JM--Osuna-C--2005"><span class="Hyperlink">Bouzouf et al., 2005</span></a>), oxidative stress (<a href="#Delvin-A--Singlh-R--Wads-E--Innes-S--Bottiglieri-T--Lentz-S--2007-"><span class="Hyperlink">Delvin et al., 2007</span></a>; <a href="#Vanzin-CS--Biancini-GB--Sitta-A--Wayhs-CA--Pereira-IN--Rockenbach-F--Garcia-SC--Wyse-AT--Schwartz-IV"><span class="Hyperlink">Vanzin et al., 2011</span></a>), cardiovascular commitment (<a href="#Refsum-H--Ueland-PM--Nygard-O--Vollset-SE--1998-"><span class="Hyperlink">Refsum et al., 1998</span></a>; <a href="#Tounyz-R--Schiffrin-E--2008"><span class="Hyperlink">Tounyz and Schiffrin, 2008</span></a>), Alzheimer disease (<a href="#Hooshmand-B--Polvikoski-T--Kivipelto-M--Tanskanen-M--Myllykangas-L--et-al.--2013"><span class="Hyperlink">Hooshmand et al., 2013</span></a>) and depression (<a href="#Nabi-H--Bochud-M--Glaus-J--Lasserre-AM--Waeber-G--et-al.--2013-"><span class="Hyperlink">Nabi et al., 2013</span></a>). Moreover, Hcy has been shown to affect lipid metabolism, which in turn contributes to the development of atherosclerosis (<a href="#Zhang-D--Fang-2012"><span class="Hyperlink">Zhang et al., 2012</span></a>).</p>
		  <p class="Caps-on-First-Para">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><span class="CharOverride-4">Punicagranatum L.</span> is used from ancient times and reports of its therapeutic qualities has echoed throughout the ages. The pomegranate fruit have many functional and medicinal effects. Pomegranate flowers attenuate aging-mediated undesirable skin abnormalities (<a href="#Wang-J--Rong-X--Um-IS--Yamahara-J--Li-Y--2012"><span class="Hyperlink">Wang et al., 2012</span></a>), besides possessing potent antioxidant and hepatoprotective activity (<a href="#Kaur-G--Jabbar-Z--Athar-M--Alam-MS--2006"><span class="Hyperlink">Kaur et al., 2006</span></a>) hypoglycemic (<a href="#Huang-RF--Hsu-YS--Lin-HI--Yang-F--2001"><span class="Hyperlink">Huang et al., 2005</span></a>)&#160;properties&#160; and cardioprotective effect (<a href="#Mohan-M--Patankar-P--Ghadi-P--Kasture-S--2010"><span class="Hyperlink">Mohan et al., 2010</span></a>; <a href="#Al-Okaily-BN--Abdullah-LN--2014-"><span class="Hyperlink">Al-Okaily and Abdullah, 2014</span></a>). Whereas, the pomegranate peel has been shown to have anti-inflammatory (<a href="#Bachoual-R--Talmoudi-W--Boussetta-T--Braut-F--El-Benna-J--2011"><span class="Hyperlink">Bachoual et al., 2011</span></a>), anti-mutagenic&#160;(<a href="#Zahin-M--Aqil-F--Ahmad-I--2010-"><span class="Hyperlink">Zahin et al., 2010</span></a>)&#160;and antifungal activity (<a href="#Endo-EH--Cortez-DA--Ueda-Nakamura-T--Nakamura-CV--Filho-BP--2010-"><span class="Hyperlink">Endo et al., 2010</span></a>). Pomegranate seeds are by-products of pomegranate juice (PJ) and they can be useful for food applications as a functional agent (<a href="#Tehranifar-AM--Zarei-Z--Nemati-B--Diyari-E--Vazifeshenas-MR--2010-."><span class="Hyperlink">Tehranifar</span><span class="Hyperlink"> et al., 2010</span></a>; <a href="#Mohagheghi-M--Rezaei-K--Labbafi-M--Mousavi-SME--2011"><span class="Hyperlink">Mohagheghi et al., 2011</span></a>). Recent studies found that pomegranate seed have been regarded as a good source of nutrients and antioxidants (<a href="#Aviram-M--Dornfeld-2000"><span class="Hyperlink">Aviram 2000</span></a>). It has been suggested that dietary supplementation with pomegranate seeds may reduce weight gain and risk of type 2 diabetes (<a href="#Strohacker-KA--Kueht-ML--2009-"><span class="Hyperlink">Strohacker and Kueht., 2009</span></a>), prevention of cancer (<a href="#Lansky-EP--Newman-RA--2007"><span class="Hyperlink">Lansky and Newman., 2007</span></a>) and also reduce menopausal symptoms (<a href="#Auerbach-L--Rakus-J--Bauer-C--et-al.--2012"><span class="Hyperlink">Auerbach et al., 2012</span></a>). The health effects of pomegranate seeds may be related to the presence of a variety of biologically active compounds (<a href="#Viuda-Martos-MJ--Fern-andez-L-opez-J--Perez--Alvarez-JA--2010"><span class="Hyperlink">Viuda-Martoset al., 2010</span></a>), particularly polyphenols (<a href="#Bachoual-R--Talmoudi-W--Boussetta-T--Braut-F--El-Benna-J--2011"><span class="Hyperlink">Balasundram et al., 2006</span></a>). Besides, significant levels of phenolic content were detected in pomegranate seeds (<a href="#Pande-G--Akoh-CC--2009"><span class="Hyperlink">Pande et al., 2009</span></a>; <a href="#Elfalleh-W--Tlili-N--Nasri-N--Yahia-Y--Hannachi-H--Chaira-N--2011-."><span class="Hyperlink">Elfalleh et al., 2011</span></a>).Therefore, this study was aimed to evaluate the protective role of pometon (PSO) against deleterious effect induced by methionine overload on oxidant/antioxidant status in adult rabbits.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">MATERIAS AND METHODS</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text">Thirty two 6-8 months female local rabbit weighing 1250-2000gm were used in this study. Animals were housed in cages in conditioned room (22-25<span class="CharOverride-6">°</span>C) in the animal house of College of Veterinary Medicine -University of Baghdad, the experiment commenced from November 2012 till February 2013. Animals had free access to water and standard pellet diet consisting according to Lab Diet Rodent Diet 5001 (PMI Nutrition International, St. Louis, MO, USA).<span class="CharOverride-2"> </span>Rabbits were randomly divided into four equal groups and treated for 42 days. Group(C):<span class="CharOverride-2"> </span>administered ordinary corn oil orally, serving as control, group T<span class="CharOverride-3">1</span> were administered methionine 100 mg/kg B.W (Segma), group<span class="CharOverride-2"> </span>T<span class="CharOverride-3">2</span> treated with same dose of methionine plus pmetone (PSO) 30mg/kg BW (vitane pharma Gmph-Germany) and group T<span class="CharOverride-3">3</span> were treated with PSO alone at same dose in T<span class="CharOverride-3">2</span>. Fasting blood samples were collected at 0, 21, 42, days of the experiment by cardiac puncture technique (cardiocentesis) by using vacuum tube with gel colt activator. Isolated serum was collected and kept at -20<span class="CharOverride-6">°</span>C until analysis for measuring the concentration of: serum total cholesterol (TC)<span class="CharOverride-2"> </span>using enzymatic kit (Linear Chemicals S.L,Spain), malondialdehyde (MDA) was determined according to (<a href="#Guidet-B--Shah-SV--1989"><span class="Hyperlink">Guidet and Shah,1989</span></a>), peroxynitrite radical (<a href="#Van-Uffelen-BE--Van-der-Zee-J--deKoster-BM--VanStereninck-J--Elferink-JG--1998-"><span class="Hyperlink">Van Uffelen et al., 1998</span></a>) and reduced glutathione (GSH) was determined by method as described by <a href="#Burtis-CA--Ashood-ER--1999-"><span class="Hyperlink">Burtis and Ashood (1999)</span></a>. Besides, serum lactate dehydrogenase (LDH) activity according to (<a href="#King-J--1965"><span class="Hyperlink">King, 1965</span></a>). The concentration of serum GSH micromole/litre was obtained from the standard glutathione curve (<a href="#Figure-1-"><span class="Hyperlink">Figure 1</span></a>).</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Heading-2--History-in-MM-">Statistical Analysis </p>
			<p class="Body-Text ParaOverride-1">The results are expressed as mean ±SE. Analysis of data was performed on the basis of Two-Way Analysis of Variance (ANOVA) using a significant level of (P&lt;0.05). Specific group differences were determined using Least Significant Differences (LSD) (<a href="#Snedecor-GW--Cochran-WG--1973-"><span class="Hyperlink">Snedecor and Cochran, 1973</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Results and Discussion&#9;</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1"><a href="#Table-1-"><span class="Hyperlink">Table 1</span></a> showed a general trend for the TC values to increase significantly (P&lt;0.05) after 21and 42days in group T<span class="CharOverride-3">1</span> compared with groups C, T<span class="CharOverride-3">2</span> and T<span class="CharOverride-3">3</span>.On the other hand, serum TC concentration dropped significantly (P&lt;0.05) at day 42 of the experiment in groups T<span class="CharOverride-3">2</span> compared to group T<span class="CharOverride-3">1</span>. Besides, PSO intubation to normal rabbits in group T<span class="CharOverride-3">1</span> caused significant decrease in this parameter along the experimental period comparing to others group.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			
            <div class="pt" > <a href="http://nexusacademicpublishers.com/uploads/figures/20150609023801.png" target="new"><img class="img_display" src="http://nexusacademicpublishers.com/uploads/figures/20150609023801.png" width="80" height="80"></a>
            
            <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-2"><a id="Figure-1-"></a>Figure 1: </span><a href="http://nexusacademicpublishers.com/uploads/figures/20150609023801.png">Standard curve of GSH</a></p>
       </div>

			
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-2"><a id="Table-1-"></a>Table 1: </span>Effect of pometone on serum total cholesterol (mg/dl) in methionine overload female rabbits</p>
			<table width="657" height="119" class="Table-Style-1" id="table-1">
				<colgroup>
					<col />
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					<col />
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					<tr class="_idGenTableRowColumn-1">
						<td height="21" colspan="4">
							<p class="Basic-Paragraph"><span class="Title CharOverride-7">Groups</span></p>
						</td>
						<td rowspan="2">
							<p class="Basic-Paragraph"><span class="Title CharOverride-7">Day</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T3</span><span class="Title CharOverride-9">(methionine +PSO)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T2 </span><span class="Title CharOverride-9">(PSO30 mg/kg BW)</span><span class="Title CharOverride-8"> </span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T1 </span><span class="Title CharOverride-9">(methionine 100 mg/kg BW)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">Control</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">93.44±0.6 </span><span class="Title CharOverride-8">A        a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">94.34±0.9 </span><span class="Title CharOverride-8">A          c</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">93.92±1.1 </span><span class="Title CharOverride-8">A        c</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">93.97±0.7 </span><span class="Title CharOverride-8">A        a            </span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">84.35±0.9 </span><span class="Title CharOverride-8">D         b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">110.84±0.4 </span><span class="Title CharOverride-8">B          b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">116.87±0.5 </span><span class="Title CharOverride-8">A          b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">94.51±0.6 </span><span class="Title CharOverride-8">C      a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">21</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">66.21±0.5 </span><span class="Title CharOverride-8">D         c</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">127.91±0.5 </span><span class="Title CharOverride-8">B            a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">183.50±0.6 </span><span class="Title CharOverride-8">A          a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">95.48±0.8 </span><span class="Title CharOverride-8">C         a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">42</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-10">Values are mean ± SE, n=8; Small letters denote within group difference p&lt;0.05. Capital letters denote between groups difference p&lt;0.05.</span></p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The results showed that oral administration of rabbits to methionine overload for six weeks (group T<span class="CharOverride-3">1</span>) caused pronounced increase in serum cholesterol concentration comparing to control and other treated groups (T<span class="CharOverride-3">2</span>, T<span class="CharOverride-3">3</span>).These results are in agreement with other studies (<a href="#Delvin-A--Singlh-R--Wads-E--Innes-S--Bottiglieri-T--Lentz-S--2007-"><span class="Hyperlink">Delvin et al.,</span><span class="Hyperlink"> 2007</span></a>; <a href="#Ventura-P--Panini-R--Verlato-C--Scarrpeta-G--Salvioli-G--2000"><span class="Hyperlink">Ventura et al., 2000</span></a>). HHcy plays an important role in cholesterol biosynthesis by inducing transcription as well as translation of 3-hydroxy 3-methyglutaryl co-enzyme reductase. It also increase cholesterol synthesis and intestinal absorption which suppressed cholesterol excretion (<a href="#Ventura-P--Panini-R--Verlato-C--Scarrpeta-G--Salvioli-G--2000"><span class="Hyperlink">Ventura et al., 2000</span></a>). Besides elevated Hcy may impaired nitric oxide (NO) through depression in plasma cysteine (<a href="#Leoccini-G--Pascale-R----Signorello-M-2003-"><span class="Hyperlink">Leoccini et al., 2003</span></a> and <a href="#Tanriverdi--H--Eurengul--H--Enli--Y--Kuru--O--Seleci--D--Tanriverdi---S--Tuzun--N--Kaftan-HA---Karab"><span class="Hyperlink">Tanriverdi et al., 2007</span></a>), the most important amino acid for cholesterol excretion, and salt formation (<a href="#Finkleston-T--Holbrok-N--2000"><span class="Hyperlink">Finkleston and Holbrook, 2000</span></a>). Accordingly, Hcy after methionine overload could be attributed to hyperlipidemia observed in this study. Hcy can potentially induced endoplasmic reticulum (ER) stress, activates and induce expression of sterol regulatory binding proteins-1, an ER membrane bound transcription factors of vascular endothelium and human hepatocytes, leading to elevate in biosynthesis and uptake of cholesterol , triglycerides with accumulation of cholesterol (<a href="#Werstuck-G--Lentz-S--Dayal-H--Hossain-G--Sood-L--shi-Y--Zhou-J--Krisans-S--Maliow-M--Austin-R--2001-"><span class="Hyperlink">Werstuk et al., 2001</span></a>; <a href="#Woo-C--Siowmy-L--Pierce-G--Choy-P--Minuk-G--Mymin-D--2005"><span class="Hyperlink">Woo et al., 2005</span></a>). The high capability of PJ to scavenge free radicals, inhibit LDL oxidation <span class="CharOverride-4">in vivo</span> and <span class="CharOverride-4">in vitro </span>(<a href="#Gil-MI---Tom-s-Barber-n--FA---Hess-Pierce--B--Holcroft--DM--Kader-AA--2000"><span class="Hyperlink">Gil</span><span class="Hyperlink"> et al., 2000</span></a>; <a href="#Aviram-M--Dornfeld-2000"><span class="Hyperlink">Aviram et al., 2000</span></a> and <a href="#Aviram-M--Dornfeld-2002"><span class="Hyperlink">2002</span></a>) and reduction in oxidative stress (<a href="#Salonen-J--Nyyssonen-2000"><span class="Hyperlink">Salonen et al., 2000</span></a> and <a href="#Salonen-RM--Nyyssonen-2003"><span class="Hyperlink">2003</span></a>)may be due to high content of polyphenols which responsible for its hypolipidemic effect.</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-2"><a id="Table-2-"></a>Table 2: </span>Effect of pometone on serum malondialdehyde concentration in methionine overload female rabbits</p>
			<table width="657" height="119" class="Table-Style-1" id="table-2">
				<colgroup>
					<col />
					<col />
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td height="21" colspan="4">
							<p class="Basic-Paragraph"><span class="Title CharOverride-7">Groups</span></p>
						</td>
						<td rowspan="2">
							<p class="Basic-Paragraph"><span class="Title CharOverride-7">Day</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T3 </span><span class="Title CharOverride-9">(methionine +PSO)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T2 </span><span class="Title CharOverride-9">(PSO30mg/ kg. BW)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8"> T1 </span><span class="Title CharOverride-9">(methionine 100 mg/kg BW)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">Control</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.26±0.004 </span><span class="Title CharOverride-8">A         c</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.25±0.003 </span><span class="Title CharOverride-8">A          c</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.25±0.003 </span><span class="Title CharOverride-8">A          c</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.25±0.004 </span><span class="Title CharOverride-8">A          a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.23±0.004 </span><span class="Title CharOverride-8">C           b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.30±0.005 </span><span class="Title CharOverride-8">B          b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.36±0.007 </span><span class="Title CharOverride-8">A           b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.26±0.005 </span><span class="Title CharOverride-8">D       a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">21</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.24±0.003 </span><span class="Title CharOverride-8">C       a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.38±0.004 </span><span class="Title CharOverride-8">B          a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.45±0.007 </span><span class="Title CharOverride-8">A        a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.25±0.005 </span><span class="Title CharOverride-8">C         a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">42</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-10">Values are mean ± SE, n=8; Small letters denote within group difference p&lt;0.05. Capital letters denote between groups difference p&lt;0.05.</span></p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Statistical analysis of the results of MDA concentration at day 21 and 42 showed that intubation of animals with methionine (group T<span class="CharOverride-3">1</span>) caused significant (p&lt;0.05) increase compared to control, T<span class="CharOverride-3">1 </span>and T<span class="CharOverride-3">3</span> (<a href="#Table-2-"><span class="Hyperlink">Table 2</span></a>). Whereas, treatment of rabbits with methionine plus PSO (group T<span class="CharOverride-3">2</span>) caused a significant (p&lt;0.05) decrease in the serum MDA concentration at the same periods of treatment compared to group T<span class="CharOverride-3">1</span>. PSO cause lowering in serum MDA concentration in group T<span class="CharOverride-3">3</span> and the results near that of control.<span class="CharOverride-2"> </span>The result showed that intubation of rabbits with methionine (group T<span class="CharOverride-3">1</span>) caused a significant (p&lt;0.05) increase in serum peroxynitrite concentration at two treated periods compared to control, T<span class="CharOverride-3">2</span>, and T<span class="CharOverride-3">3</span> groups. Results have also clarified that treatment of rabbits with PSO (group T<span class="CharOverride-3">3</span>) showed significant (p&lt;0.05) decrease in the serum peroxynitrite concentration at 21 and 42 days of experiment compared to T<span class="CharOverride-3">1</span> group. Highest significant (p&lt;0.05) reduction in serum peroxynitrite concentration were observed after pomegranate seed oil intubation to normal animals in group T<span class="CharOverride-3">3</span> comparing to T<span class="CharOverride-3">1</span> and T<span class="CharOverride-3">2</span> treated groups (<a href="#Table-3-"><span class="Hyperlink">Table 3</span></a>). </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">A significant (p&lt;0.05) decrease in serum GSH concentration were detected after 42 days of experiment in groups T<span class="CharOverride-3">1</span> and T<span class="CharOverride-3">2</span> comparing to control and T<span class="CharOverride-3">3</span> treated groups (<a href="#Table-4-"><span class="Hyperlink">Table 4</span></a>). The results also showed that oral intubation of PSO to normal animals caused significant (p&lt;0.05) increase in serum GSH concentration of treated rabbits (group T<span class="CharOverride-3">3</span>) comparing to other treated. </p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-2"><a id="Table-3-"></a>Table 3: </span>Effect of pometone on serum peroxynitrite radical concentration in methionine overload female rabbits</p>
			<table width="657" height="119" class="Table-Style-1" id="table-3">
				<colgroup>
					<col />
					<col />
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td height="21" colspan="4">
							<p class="Basic-Paragraph"><span class="Title CharOverride-7">Groups</span></p>
						</td>
						<td width="35" rowspan="2">
							<p class="Basic-Paragraph"><span class="Title CharOverride-7">Day&#9;&#9;</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td width="134" height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T3 </span><span class="Title CharOverride-8">(methionine+</span><span class="Title CharOverride-8">PSO)</span></p>
						</td>
						<td width="173">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T2 </span><span class="Title CharOverride-8">(PSO30 mg / kg. BW)</span></p>
						</td>
						<td width="200">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T1 </span><span class="Title CharOverride-8">(methionie 100 mg/kg.   B.W)</span></p>
						</td>
						<td width="91">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">Control</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">33.51±0.5 </span><span class="Title CharOverride-8">A      a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">34.72±0.4 </span><span class="Title CharOverride-8">A         c</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">33.51±0.3 </span><span class="Title CharOverride-8">A          c</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">33.57±0.7 </span><span class="Title CharOverride-8">A     a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">31.68±0.4 </span><span class="Title CharOverride-8">C       b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">40.03±0.3 </span><span class="Title CharOverride-8">Bb</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">42.43±0.4 </span><span class="Title CharOverride-8">A         b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">32.08±0.4 </span><span class="Title CharOverride-8">Ca</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">21</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">32.68±0.5 </span><span class="Title CharOverride-8">C     b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">45.03±1.03 </span><span class="Title CharOverride-8">B          a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">63.25±1.01 </span><span class="Title CharOverride-8">A          a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">33.50±0.3 </span><span class="Title CharOverride-8">C       a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">42 </span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-10">Values are mean ± SE, n=8; Small letters denote within group difference p&lt;0.05. Capital letters denote between groups difference p&lt;0.05.</span></p>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-2"><a id="Table-4-"></a>Table 4: </span>Effect of pomegranate seed oil on serum reduced glutathione concentration in methionine overload female rabbits</p>
			<table width="658" height="138" class="Table-Style-1" id="table-4">
				<colgroup>
					<col />
					<col />
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td height="21" colspan="4">
							<p class="Basic-Paragraph"><span class="Title CharOverride-7">Groups</span></p>
						</td>
						<td width="41" rowspan="2">
							<p class="Basic-Paragraph"><span class="Title CharOverride-7">Day</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td width="138" height="40">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T3 </span><span class="Title CharOverride-8">(methionine+ </span><span class="Title CharOverride-8">PSO)</span></p>
						</td>
						<td width="168">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T2 </span><span class="Title CharOverride-8">(PSO30 mg / kg. BW)</span></p>
						</td>
						<td width="174">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T1 </span><span class="Title CharOverride-8">(methionine  </span><span class="Title CharOverride-8">100 mg / kg. BW)</span></p>
						</td>
						<td width="113">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">Control</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.05±0.0010 </span><span class="Title CharOverride-8">A           c</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.06±0.0007 </span><span class="Title CharOverride-8">A           a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.06±0.0007 </span><span class="Title CharOverride-8">A           a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.06±0.0008 </span><span class="Title CharOverride-8">A          a               </span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.13±0.0006 </span><span class="Title CharOverride-8">A         b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.05±0.0006 </span><span class="Title CharOverride-8">D     b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.05±0.0010 </span><span class="Title CharOverride-8">C           b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.06±0.0007 </span><span class="Title CharOverride-8">B           a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">21</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.14±0.0010 </span><span class="Title CharOverride-8">A         a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.05±0.0004 </span><span class="Title CharOverride-8">C         b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.05±0.0005 </span><span class="Title CharOverride-8">D              b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0.06±0.0006 </span><span class="Title CharOverride-8">B            a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">42</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Normal" xml:lang="en-US">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-10">Values are mean ± SE, n=8; Small letters denote within group difference p&lt;0.05. Capital letters denote between groups difference p&lt;0.05.</span></p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The overall results of the present study demonstrated that treatment of rabbits with methionine overload induced an elevation in oxidative stress parameters, evidenced by increased serum MDA and peroxynitrate and decreased reduced GSH concentrations. Biological oxidative stress is defined by an imbalance between antioxidant and free radicals, and this phenomenon is associated with HHcy (methionine overload) (<a href="#Petrak-D--Myslivcva-P--Man-R--Cmejla-J--Vylor-M--Elleder-V--Vulp-C--2007-"><span class="Hyperlink">Petrak et al., 2007</span></a>; <a href="#Tounyz-R--Schiffrin-E--2008"><span class="Hyperlink">Tounyz and Schifrin, 2008</span></a>). It has well known that the free radicals initiate the peroxidation of membrane polyunsaturated fatty acids which results in the generation and excess production of reactive ROS and finally cell necrosis (<a href="#Melin-AM--Perromat-A--Deleris-G--2000-."><span class="Hyperlink">Melin et al., 2000</span></a>; <a href="#Manna-P--Bhattacharyya-S--Das-J--Ghosh-J--Sil-P--2011-."><span class="Hyperlink">Manna et al., 2011</span></a>). The subsequent increase in serum MDA concentration in the current study may be attributed to high sensitivity of rabbits to free radicals production (by HHcy) induced by methionine over load. This elevation in generation of lipid peroxidation (MDA level) was postulated to cause a gradual cell injury by free radicals liberating lipoxygenase enzymes which oxidized unsaturated membrane fatty acids and subsequent production of MDA, overwhelming endogenous scavenging system including GSH resulting in oxidative stress (<a href="#Stark-G--2005"><span class="Hyperlink">Stark, 2005</span></a>; <a href="#Handy-D--Zhang-L--2006-"><span class="Hyperlink">Handy and Zhang, 2006</span></a>) .</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">It has been postulated that peroxynitrite mediated one-electron oxidation of biomolecules may be an important event in its cytotoxic mechanism (<a href="#Augusto-O--Gatti-RM--Radi-R--1994"><span class="Hyperlink">Augusto et al., 1994</span></a>). The generation of peroxynitrate over long periods of time will result in substantial oxidation of potential destruction of host cellular constituents, leading to the dysfunction of critical cellular processes, disruption of cell signalling pathway, and the induction of cell death through both apoptosis and necrosis (<a href="#Pacher-P--Beckman-JS--Liaudet-L--2007-."><span class="Hyperlink">Pacher et al., 2007</span></a>).</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">A significant decrease in serum GSH concentration in methionine treated group in the present study is correlated with (<a href="#Ventura-P--Panini-R--Verlato-C--Scarrpeta-G--Salvioli-G--2000"><span class="Hyperlink">Ventura et al., 2000</span></a>; <a href="#Huang-RF--Hsu-YS--Lin-HI--Yang-F--2001"><span class="Hyperlink">Huang et al., 2001</span></a>; <a href="#Hayden-M--Tyage-H--2004"><span class="Hyperlink">Hayden and Tyage, 2004</span></a>).<span class="CharOverride-13"> </span>Many authors demonstrated that mild HHcy is much more common and is associated with postmethionine loading in water (<a href="#Labinjoh-C--Newby-DE--Wilkinson-IB--Megson-IL--MacCallum-H--Melville-V--Boon-NA--Webb-DJ--2001"><span class="Hyperlink">Labinjoh et al., 2001</span></a>; <a href="#Bagi-Z--Cseko-C--Toth-R--Koller-A--2003-"><span class="Hyperlink">Bagi et al., 2003</span></a>) or in diet (<a href="#Apeland-T--Mansoor-MA--Pentieva-K.MC--Nulty-H--Seljeflot-I--Strand-jord-RE--2003"><span class="Hyperlink">Apeland et al., 2003</span></a>; <a href="#Zhang-R--Ma-J--Zhu-H--Ling-W--2004"><span class="Hyperlink">Zhang et al., 2004</span></a>). The predicted HHcy after methionine overload may decrease the ability of the cells to detoxify H<span class="CharOverride-3">2</span>O<span class="CharOverride-3">2</span> and other lipid peroxide (<a href="#Tounyz-R--Schiffrin-E--2008"><span class="Hyperlink">Tounyz and Schifrin, 2008</span></a>) and it might be indirectly produce oxidative reduction in the activity of intracellular antioxidant enzyme. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Body-Text">The present study showed that PSO supplementation during treatment of methionine overload exhibited an increase in GSH concentration and reduce MDA and peroxynitrate concentration, compared to methionine–treated group (T<span class="CharOverride-3">1</span>). Phenolic compound (<a href="#Kaur-G--Jabbar-Z--Athar-M--Alam-MS--2006"><span class="Hyperlink">Kaur</span><span class="Hyperlink"> et al., 2006</span></a>) present in juice , including punicalagin, ellagic acid (<a href="#Cuzzocree--S--Riely-DP--Coupli-AP--Salvemini--M--2001"><span class="Hyperlink">Cuzzocree et al., 2001</span></a>; <a href="#Chen-B--Tuuli-MG--Longtine-MS--Shin-JS--Lawrence-R--Inder-T--Michael-ND--2012-."><span class="Hyperlink">Chen et al., 2012</span></a>) and anthocyanins (<a href="#Seeram-NP--Adams-LS--Henning-SM--Niu-Y--Zhang-Y--Nair-MG--Heber-D--2005-"><span class="Hyperlink">Seeram et al., 2005</span></a>)<span class="CharOverride-2"> </span>are known for their properties in scavenging free radicals and inhibiting lipid oxidation and elevation antioxidants capacity of the body. The mechanisms of antioxidant effects of PJ may include: 1)<span class="CharOverride-2"> </span>suppressing reactive oxygen species formation either by inhibition of enzymes or chelating trace elements involved in free radical production, 2) scavenging reactive oxygen species, 3) up regulating or protecting antioxidant defenses<span class="CharOverride-2"> </span>(<a href="#Grassi-D--Desideri-2009a"><span class="Hyperlink">Grassi et al., 2009a</span></a>; <a href="#Grassi-D--Desideri-2009b"><span class="Hyperlink">Grassi et al., 2009b</span></a>; <a href="#Ferri-C--Grassi-D--2010-."><span class="Hyperlink">Ferri and Grassi, 2010</span></a>) and 4) increasing resistance to oxidative stress (<a href="#Li-Y--Guo-C--Yang-J--Wei-J--Xu-J--Cheng-S--2006-"><span class="Hyperlink">Li et al</span><span class="Hyperlink">., 2006</span></a>; <a href="#Bachoual-R--Talmoudi-W--Boussetta-T--Braut-F--El-Benna-J--2011"><span class="Hyperlink">Bachoual et al., 2011</span></a>).</p>
		  <p class="Body-Text">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Significant decrease in LDH activity was shown in T<span class="CharOverride-3">2</span> treated group comparing to group T<span class="CharOverride-3">1</span> at 21 and 42 days of the experiment (<a href="#Table-5-"><span class="Hyperlink">Table 5</span></a>). Highest significant (p&lt;0.05) increment in this parameters was shown at the end of experiment in methionine treated group (T<span class="CharOverride-3">1</span>), methionine plus PSO (T<span class="CharOverride-3">2</span>) comparing to control and group T<span class="CharOverride-3">3</span>. While intubation of pomegranate seed oil to normal animals (T<span class="CharOverride-3">3</span>) caused significant (p&lt;0.05) decrement in serum LD activity comparing to T<span class="CharOverride-3">1</span> and T<span class="CharOverride-3">2</span> treated groups. Also the results have clarified that were no significant (p&gt;0.05) differences between T<span class="CharOverride-3">3</span> and control group in the same duration of treatment when compared in each other.</p>
			<p class="Body-Text ParaOverride-1">&nbsp;</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-2"><a id="Table-5-"></a>Table 5: </span>Effect of pometone on serum lactate dehydrogenase activity in methionine overload female rabbits</p>
			<table width="658" height="120" class="Table-Style-1" id="table-5">
				<colgroup>
					<col />
					<col />
					<col />
					<col />
					<col />
				</colgroup>
				<tbody>
					<tr class="_idGenTableRowColumn-1">
						<td height="21" colspan="4">
							<p class="Basic-Paragraph"><span class="Title CharOverride-7">Groups</span></p>
						</td>
						<td rowspan="2">
							<p class="Basic-Paragraph"><span class="Title CharOverride-7">Day</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td height="22">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T3 </span><span class="Title CharOverride-8">(methionine+ PSO)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T2 </span><span class="Title CharOverride-8">(PSO30 mg/ kg. BW)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">T1 </span><span class="Title CharOverride-8">(methionine </span><span class="Title CharOverride-8">100 mg /kg. BW)</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">Control</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">63.81±0.8 </span><span class="Title CharOverride-8">A          a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">65.84±1.2 </span><span class="Title CharOverride-8">A             c</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">65.87±0.9 </span><span class="Title CharOverride-8">A            c</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">65.25±1.06 </span><span class="Title CharOverride-8">A           a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">0</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-2">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">62.61±1.05 </span><span class="Title CharOverride-8">C          a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">104.73±0.9 </span><span class="Title CharOverride-8">B        b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">137.28±0.9 </span><span class="Title CharOverride-8">A        b</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">64.88±1.08 </span><span class="Title CharOverride-8">C            a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">21</span></p>
						</td>
					</tr>
					<tr class="_idGenTableRowColumn-1">
						<td height="21">
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">65.13±1.04 </span><span class="Title CharOverride-8">C       a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">114.92±0.9 </span><span class="Title CharOverride-8">B        a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">158.96±0.9 </span><span class="Title CharOverride-8">A         a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">64.93±1.2 </span><span class="Title CharOverride-8">C          a</span></p>
						</td>
						<td>
							<p class="Basic-Paragraph"><span class="Title CharOverride-8">42</span></p>
						</td>
					</tr>
				</tbody>
			</table>
			<p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Figure--and-Table-Heading ParaOverride-1"><span class="CharOverride-10">Values are mean ± SE, n=8; Small letters denote within group difference p&lt;0.05. Capital letters denote between groups difference p&lt;0.05.</span></p>
		  <p class="Figure--and-Table-Heading ParaOverride-1">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The present study showed a significant elevation in LDH activity in animals treated with methionine for six weeks indicating occurrence of liver function disorder (<a href="#Ferre-N--Gomez-F--Camps-J--Simo-J--Michelle-M--Ballart-J--Joven-J--2002"><span class="Hyperlink">Ferre et al., 2002</span></a>; <a href="#Ebbesen-S--Ingersity-J--2005-"><span class="Hyperlink">Ebbesen and Ingersity, 2005</span></a>). The liver produce a large amount of LDH which are secreted to the circulation with injury or death (<a href="#Wolff-SP--2006-"><span class="Hyperlink">Wolff, 2006</span></a>), in addition, release of liver enzyme from cytosol can occur secondary to cellular necrosis with membrane damage (<a href="#Valentin-BA--Blue-JT--Shelley-SM--Cooper-BJ--1990"><span class="Hyperlink">Valentin et al., 1990</span></a>). This fact is documented in this study by the oxidative stress induced by methionine overload leading to cell damage and increase in this enzyme. Also methionine over load may cause oxidative stress and damaging the hepatocell membrane and central portal liver cells with subsequent release of AST and ALT enzymes (<a href="#Garcia-Terijiano-E--Barasia-C--Rodriguez-J--2001"><span class="Hyperlink">Garcia-Terijano et al., 2001</span></a>; <a href="#Sahi-A--Pan-X--Paul-R--Malladi-P--Kahli-R--Whitington-F--2006"><span class="Hyperlink">Sahi et al., 2006</span></a>). The current study found a marked significant decrease in LDH activity after PSO intubation explained the alleviation of oxidative insults by antioxidants properties of PSO. Some studies showed the presence of flavonoids, steroids, terpenoids and tannins in pomegranate reduce xenobiotic- induced hepatotoxicity in animals and counteract the damaging effects of oxidative stress, cooperating with natural system like glutathione and other endogenous protective enzyme (<a href="#Abdel-Rahman-MK--Abdel-Megeid-AA--2006-"><span class="Hyperlink">Abdel-Rahman and Abdel-Megeid, 2006</span></a>; <a href="#Toklu-HZ--Dumlu-MU--Sehirli-O--Ercan-F--Gedik-N--Gokmen-V--Sener-G--2007-"><span class="Hyperlink">Toklu et al., 2007</span></a>). No study, however reviewed the effect of pomegranate seed oil on serum LDH as a hepatoprotective agent. In conclusion, methionine overload induced oxidative stress and pomegranate seed oil can ameliorate the oxidative stress evidenced by measured biomarkers.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">CONCLUSION</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">The study conclude that methionine overload induced oxidative stress and pomegranate seed oil can ameliorate the oxidative stress evidenced by measured biomarkers.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">CONFLICT OF INTEREST</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">There do not exist any conflict of interest.</p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Authors contribution</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Baraa Najim Al-Okaily designed the experiment, analyzed and interpreted the data. Lekaa Najim Abdulla performed the experiment and wrote the paper. Khalisa Khadim Khudair gave technical support and conceptual advice. </p>
		  <p class="Body-Text ParaOverride-1">&nbsp;</p>
			<p class="Heading-1--Introduction----">Aknowledgement</p>
		  <p class="Heading-1--Introduction----">&nbsp;</p>
			<p class="Body-Text ParaOverride-1">Authors of this study would like to thank College of Veterinary Medicine, University of Baghdad, Iraq for their support.</p>
			<p class="Heading-1--Introduction----">&nbsp;</p>
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