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Rice Bran Oil and Pumpkin Seed Oil Alleviate Oxidative Injury and Fatty Liver in Rats Fed High Fructose Diet

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Rice Bran Oil and Pumpkin Seed Oil Alleviate Oxidative Injury and Fatty Liver in Rats Fed High Fructose Diet Pol. J. Food Nutr. Sci., 2014, Vol. 64, No. 2, pp. 127–133 DOI: 10.2478/pjfns-2013-0002 http://journal.pan.olsztyn.pl Original article Section: Nutrition Research Rice Bran Oil and Pumpkin Seed Oil Alleviate Oxidative Injury and Fatty Liver in Rats Fed High Fructose Diet Sahar Y. Al-Okbi1, Doha A. Mohamed1*, Thanaa E. Hamed1, Reham S.H. Esmail2 ¹Food Sciences and Nutrition Department, National Research Centre, Dokki, Cairo, Egypt ²Pathology Department, National Research Centre, Dokki, Cairo, Egypt Key words: fatty liver, non-alcoholic fatty liver with infl ammation, fructose, rice bran oil, pumpkin seed oil Nonalcoholic steatohepatitis (NASH) and nonalcoholic fatty liver disease are increasing in adults and are likely to be increasing in children. The aim of the present research was to evaluate the protective effect of rice bran oil and pumpkin seed oil against high fructose diet (HFD) inducing nonalcoholic steatohepatitis (NASH). The results showed signifi cant elevation of plasma total and direct bilirubin, transaminases activities, total cholesterol (T-Ch), triglycerides (TG), low density lipoprotein-cholesterol (LDL-Ch), tumor necrosis factor- (TNF-) and malondialdehyde (MDA) with signifi cant increase in liver TG, T-Ch and MDA along with signifi cant reduction in plasma high density lipoprotein cholesterol (HDL-Ch) and in- crease in T-Ch/HDL-Ch in rats fed HFD compared to rats fed on balanced diet. Histopathology of liver of rats fed on HFD confi rmed the induction of NASH. Rice bran oil and pumpkin seed oil produced improvement in the biochemical parameters with different degrees. Pumpkin seed oil reversed all histopathological changes that occur in liver tissue which became comparable to normal in some rats. In conclusion, rats fed high fructose diet are a good model for studying NASH. Rice bran oil and pumpkin seed oil afford hepato protection against NASH in rat model. INTRODUCTION NAFLD is the most common cause of chronic liver injury worldwide. It has a broad pathologic spectrum which ranges Fructose is a monosaccharide that is widely available from simple fatty infi ltration of the liver or steatosis, to non- in natural food sources such as fruits and honey. However, alcoholic steatohepatitis (NASH), fi brosis, cirrhosis and to in most countries the main source of fructose is from su- liver failure [Assy et al., 2000]. NASH is fatty liver with in- crose, a disaccharide composed of equal portions of fructose fl ammation and elevated oxidative stress and it is a progres- and glucose. Fructose intake has increased markedly over sive form of NAFLD that is diagnosed by histopathological the last 2 centuries, primarily due to the increasing intake features [Brunt et al., 1999]. NAFLD and NASH have been of sucrose and high-fructose corn syrup [Tappy & Le, 2010; recently recognized as hepatic manifestations of metabolic Ha et al., 2013]. Fructose is known to stimulate fat accumu- syndrome [Day & Saksena, 2002]. It has been shown pre- lation in the liver by both increasing synthesis and block- viously that experimental NASH is accompanied by liver ing fat oxidation [Ackerman et al., 2005]. Therefore, fruc- dysfunction and dyslipidemia and that NASH is a risk factor tose and sugar-sweetened beverages have been related to for cardiovascular diseases [Al-Okbi et al., 2013]. So, it can the risk of non-alcoholic fatty liver disease (NAFLD). Plas- be hypothesized that therapy which improves liver function ma triglycerides are increased by sugar-sweetened beverages, and produces antioxidant, anti-infl ammatory, hypolipidemic and this increase appears to be due to fructose moiety [Bray, and lipotropic effect might prevent progression of NASH 2012]. NAFLD is the most common hepatic manifestation and its cardiovascular risk. Based on lack of effective ther- of obesity, affecting 20%–30% of adults [Neuschwander-Te- apies for NASH, natural bioactive constituents from plant tri, 2005; Vos & Lavine, 2013]. NAFLD is defi ned as the ac- materials especially plant foods might be useful in this re- cumulation of lipid, primarily triacylglycerols, in the liver spect. From such plant foods; rice bran oil and pumpkin seed [McCullough, 2004]. Soft drink consumption was associ- oil are reported to contain bioactive constituents that might ated with NAFLD independent of metabolic syndrome [Abid possess the aforementioned activities. Rice bran oil (RBO) et al., 2009] or in the absence of traditional risk factors, in- is a rich source of bioactive constituents such as oryzanols, cluding obesity, diabetes or hyperlipidemia [Assy et al., 2008]. phytosterols, tocopherols, tocotrienols, squalene, polico- In fact, an increasing body of evidence indicates that fruc- sanols and ferulic acid [Khatoon & Gopalakrishna, 2004; tose in the diet itself causes NAFLD [Nomura et al., 2012]. Ardiansyah et al., 2006]. These bioactive constituents have been reported to possess multiple health benefi ts including reduction of cholesterol levels [Chen & Cheng, 2006], anti- * Corresponding Author: Tel.: 01222357571; Fax: (202) 33370931; E-mail: [email protected] (Prof. Dr. Doha A. Mohamed) oxidant [Xu et al., 2001] and anti-infl ammatory activity [Aki- © Copyright by Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences © 2014 Author(s). This is an open access article licensed under the Creative Commons Attribution-NonCommercial-NoDerivs License (http://creativecommons.org/licenses/by-nc-nd/3.0/). 128 Alleviation of Oxidative Injury and Fatty Liver in Rats by Rice Bran and Pumpkin Oils TABLE 1. Composition of experimental diets (g/100 g). Experimental procedures Twenty-four rats were divided into four groups, each of six High High High fructose diet fructose diet Balanced rats. The fi rst was normal group where rats received a bal- Ingredients fructose containing containing diet anced diet. The second group was control where rats were fed diet rice bran oil pumpkin oil on high fructose diet (for induction of NASH). Rats of group Casein 17.0 17.0 17.0 12.0 three and four were fed on high fructose diet containing rice Corn oil - - - 10.0 bran oil and pumpkin seed oil and also given an oral daily Rice bran oil - 5.5 - - dose of 200 mg/rat/day of rice bran oil and pumpkin seed oil, respectively. During the experiment, body weight and food Pumpkin oil - - 5.5 - intake were recorded weekly. After thirty-fi ve days (end Butter fat 5.5 - - - of the study) total food intake, body weight gain and food Fructose 70.0 70.0 70.0 - effi ciency ratio (body weight gain/total food intake) were Starch - - - 70.5 calculated. Blood samples were collected from eye orbital Salt mix. 3.5 3.5 3.5 3.5 of anaesthetized rats after an overnight fast. Heparin was Vitamin mix. 1.0 1.0 1.0 1.0 added to blood and plasma was separated by centrifugation at 3000 rpm. Plasma total cholesterol (T-Ch) [Watson, 1960], Cellulose 3.0 3.0 3.0 3.0 high density lipoprotein cholesterol (HDL-Ch) [Burstein et al., 1970], low density lipoprotein cholesterol (LDL-Ch) hisa et al., 2000]. Pumpkin seed oil has been reported to pos- [Schriewer et al., 1984] and triglycerides (TG) [Megraw sess anti-infl ammatory, hypolipidemic and antioxidant effect et al., 1979] were determined. The T-Ch / HDL-Ch ratio was [Suresh & Das, 2003; Makni et al. 2008, 2010]. These re- calculated. Malondialdehyde (MDA) was determined as an ported bioactivities of both RBO and pumpkin seed oil could indicator of lipid peroxidation [Satoh, 1978]. Plasma tumor render them potential health benefi t as fatty liver preventives. necrosis factor- (TNF-) (an infl ammatory biomarker) So the aim of the present study was to evaluate the benefi cial [Stepaniak et al., 1995] was determined. The activity of as- effect of rice bran oil and pumpkin seed oil administration partate transaminase (AST) and alanine transaminase (ALT) in rats fed high fructose diet inducing NASH. NASH-related [Reitman & Frankel, 1957] was estimated as indicators of liv- biochemical parameters planned to be assessed included liv- er function and/or damage. Liver was immediately removed, er fat, plasma lipids, TNF-, MDA in both plasma and liver weighed and stored at -20°C till analyzed. Total hepatic lip- and liver function tests along with histopthological examina- ids were extracted and weighed according to the procedure tion of the liver. of Folch et al. [1957] and the concentration of triglycerides and cholesterol assessed [Megraw et al., 1979 and Watson, MATERIAL AND METHODS 1960, respectively]. MDA was determined in the liver accord- ing to Ohkawa et al. [1979]. For histopathological study, part Plant materials of liver was removed, placed in 10% formaldehyde, dehydrat- Egyptian rice bran was stabilized after milling by heating ed in graded alcohol and embedded in paraffi n. Fine sections at 125°C for 15 min and supplemented by Dr. Amr M. Helal, were prepared, mounted on glass slides and counter-stained International Trade & Marketing Managing, Cairo, Egypt. with hematoxylin and eosin for light microscopic analysis Pumpkin seed oil (Cucurbita pepo L., Family Cucurbitaceae [Ekor et al., 2010]. Animal procedures were performed in ac- var. styria) was obtained from Graz, Austria. cordance with the Ethics Committee of the National Research Centre, Cairo, Egypt, and followed the recommendations Animals of the National Institutes of Health Guide for Care and Use Male Sprague Dawley rats of body weight equal to of Laboratory Animals (Publication No. 85–23, revised 1985). 160.3±15.13 g as mean±SD were used in the present study. Animals were obtained from Animal House of National Re- Statistical analysis search Centre, Cairo, Egypt. Animals were kept individually The results of animal experiments are expressed as in stainless steel cages; water and food were given ad-libtium. the mean±SE and they are analyzed statistically using the one-way analysis of variance ANOVA followed by Dun- Preparation of plant extracts can’s test.
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