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Food and Chemical Toxicology 48 (2010) 2407–2412

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Food and Chemical Toxicology
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Flax and Pumpkin seeds mixture ameliorates diabetic nephropathy in rats
Mohamed Makni a,b, Mediha Sefi a,1, Hamadi Fetoui a,1, El Mouldi Garoui a, Nabil K. Gargouri b,
Tahia Boudawara c, Najiba Zeghal a,*
a
Animal Physiology Laboratory, Faculty of Sciences, BP 1171, 3000 Sfax, Tunisia
b
Food Processing Department, ISET, BP 377, 9100 Sidi Bouzid, Tunisia
c
Histopathology Laboratory, CHU Habib Bourguiba, 3029 Sfax, Tunisia

a r t i c l e i n f o a b s t r a c t

Article history: This study investigated the hypoglycemic and antioxidant effects of Flax and Pumpkin seeds mixture on
Received 3 April 2010 the kidney of alloxan-induced diabetic rats. Animals were allocated into three groups of six rats each: a
Accepted 28 May 2010 control group (CD), a diabetic group (DD) and diabetic rats fed with Flax and Pumpkin seeds mixture
(DMS) group. The DD rats showed a significant increase of glycemia and lipid parameters such as total
lipid, total cholesterol and triglycerides levels compared to those of the control group (CD). In addition,
Keywords: plasma and kidney malonaldialdehyde levels (MDA) were significantly increased compared to (CD)
Diabetes
group. Antioxidant enzyme activities such as catalase (CAT), superoxide dismutase (SOD) and non-enzy-
Oxidative stress
Antioxidant enzymes
matic levels of reduced glutathione (GSH) significantly decreased in the plasma and kidney of diabetic
Lipid peroxidation rats compared to those of controls. Diet supplemented with Flax and Pumpkin seeds mixture ameliorated
Flax and Pumpkin seeds the antioxidant enzymes activities observed in diabetic rats and significantly decreased MDA levels. Kid-
Nephropathy ney histological sections, showed glomerular hypertrophy and tubular dilatation. In DMS rats, these his-
topathological changes were less prominent. Our results suggest that Flax and Pumpkin seeds mixture
supplemented in diet of diabetic rats may be helpful to prevent diabetes and its complications.
Ó 2010 Elsevier Ltd. All rights reserved.

1. Introduction Ugochukwu et al., 2003; Maritim et al., 2003). Oxygen free radicals
are formed disproportionately in diabetes by glucose oxidation,
Diabetes mellitus (DM) is a health problem affecting millions of non-enzymatic protein glycation and the subsequent oxidative
individuals worldwide. The World Health Organization (WHO) pre- degradation of glycated proteins (Maritim et al., 2003).
dicts that 300 millions of people will have diabetes mellitus by the Many scientific reports indicate that diabetic complications are
year 2025 (Pradeepa and Mohan, 2002). DM is a prevalent systemic associated with overproduction of ROS and accumulation of lipid
disease with well documented devastating effects (Duckworth, peroxidation by-products (Palanduz et al., 2001). In diabetes, major
2001). damage occurs in tissues such as kidney where the entry of glucose
Hyperglycemia has been found to play a key role in reactive is not regulated by insulin (Limaye et al., 2003).
oxygen species (ROS) generated damage (Ha and Kim, 1999; Free radicals generated in diabetes may lead to several kinds of
Greenberg and Sacks, 2002; Ugochukwu and Babady, 2002; diabetic complications including nephropathy, neuropathy, cardi-
opathy and many others diseases. Many herbal extracts used as
single agents or in different oral formulations have been recom-
Abbreviations: AI, atherogenic index; CAT, catalase; CD, control rats; DD,
mended to prevent diabetes mellitus due to the fact that they are
diabetic rats; DM, diabetes mellitus; DMS, diabetic rats fed diet containing seeds less toxic than oral hypoglycemic agents such as sulfonylureas,
mixture; DTNB, 5,5-dithiobis-2 nitro benzoic acid; FCE, food conversion efficiency; metformin, etc. (Ponnachan et al., 1993; Chattopadhyay, 1993).
GSH, reduced glutathione; GTT, glucose tolerance test; HDL-C, high-density Antioxidants play a major role in protection against molecular oxi-
lipoprotein-cholesterol; LDL-C, low-density lipoprotein-cholesterol; MDA,
dative damage (Evans, 2007).
malondialdehyde; NBT, nitroblue tetrazolium; PUFAs, polyunsaturated fatty acids;
ROS, reactive oxygen species; SD, standard deviation; SOD, superoxide dismutase; Traditional medicines, derived mainly from plants, play a major
TBA, thiobarbituric acid; TBARS, thiobarbituric acid-reactive substance; TC, role in the management of diabetes mellitus (Karunanayake and
total cholesterol; TG, triacylglycerol; WHO, World Health Organization. Tennekoon, 1993; Patel and Srinivasan, 1997; Ahmed et al., 2004).
* Corresponding author. Address: Animal Physiology Laboratory, UR 08-73, Sfax World Health Organization (WHO) recommended the evalua-
Faculty of Sciences, BP 1171, 3000 Sfax, Tunisia. Tel.: +216 98 914 154; fax: +216 74
274 437.
tion of traditional plant treatments for diabetes as they are effec-
E-mail address: najiba.zeghal@tunet.tn (N. Zeghal). tive, non-toxic, with less or no side effects and are considered to
1
Authors contributed equally to this work. be excellent candidates for oral therapy (Day, 1998). Recently,

0278-6915/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.fct.2010.05.079

20131. Estimation of urea. Changes in absorbance were re- corded at 240 nm. 2008). Ariana. The contents of total lipids in plasma mixture seems to exert a protective effect against weight loss in extracts were quantified gravimetrically by evaporating off the solvents using a ro.4.5 ml of each plasma sample or kidney extract supernatant (tissue homogenates 2. No significant difference was observed in digestibility using commercial kits from Biomaghreb (Refs.5. Determination of blood glucose and glucose tolerance test (GTT) Plasma glucose levels were assayed by enzymatic methods.4. Induction of diabetes min/mg of protein. used already in traditional medicine. GTT was conducted in degree of tubular and glomerular injury and necrosis. digestibility and feed con- 2. were deter- 2. Company of Rodent Diet. crushed at ambient temperature 15 min at 90 °C and cooled. Plant material in potassium phosphate buffer pH 7. 3.6. AKRIN-010T (Shibayagi Co.4. 2. as recommended by the WHO and according to Blandeau and Schneider (2006) and using 1. diabetes was induced in male rats which re- ceived by intraperitoneal way. The developed blue color in the reaction was measured at 560 nm. Sfax. Then they were embedded in paraffin and stained with The glucose tolerance test (GTT) evaluates the ability to respond appropriately hematoxylin–eosin for histopathological studies.2. agent. pH 7. Histopathological examination 2.1. 2 lM riboflavin and 75 lM Nitroblue tet- oratory conditions (22 ± 3 °C. BAYER. RIT-461 No. Analysis of plasma lipids version efficiency (FCE) of experimental animals. experimental and clinical 1972): antidiabetic. (1957).2408 M. The low-density lipoprotein-cholesterol (LDL-C) fraction and atherogenic index (AI) were determined according to the Friedewald equations (Friedewald et al. Rats were orally treated with 20% glucose solution (5–10 ml) after 6 h. Grigg (2004).2.1 mM EDTA. 2. Tunisia). Superoxide dismutase (SOD) activ- ity was estimated according to Beauchamp and Fridovich (1971). (2002) and Sheweita et al. Those with moderate diabetes and where blood glucose con.6. 5% glucose water solution to pre. (2006) have recommended many medicinal plants sia). homogenized tissue and the substrate (H2O2) to a concentration of 0. / Food and Chemical Toxicology 48 (2010) 2407–2412 Mankil et al. 12-h light/dark cycle).4. uric acid and creatinine in plasma were estimated spectro- photometrically using commercial diagnostic kits. Ltd. Total superoxide dismutase activity (SOD). Berkley.3. 2.05. and FCE between groups.4. antidiabetic and nephroprotective effects of Flax and Pumpkin AI ¼ ðTC  HDL-CÞ=HDL-C: seeds mixture rich in PUFAs and antioxidant compounds. Germany)..5 ml of supernatant were incubated for chased from a local market (Sfax city. Statistical analysis between CD and DD. Results 2.8).6.4. be. Tunisia). buffer were added to 3 ml of 4% sulfosalicylic acid. 0. England). Tunisia). The ratio of Omega 6/Omega 3 fatty acids was 5/1 idation was expressed as nmol of thiobarbituric acid-reactive substances (TBARS). European Communities.3. an index of lipid peroxidation. 20113. respectively (Refs.3. The local Committee of Ethics in Animal Experimen- tation approved all the experimental procedures of this study. uric acid and creatinine The levels of urea. 1986).1. 20121. triacylglycerol (TG) and high-density and the food conversion efficiency (FCE) ratio were presented in lipoprotein-cholesterol (HDL-C) levels were determined by enzymatic methods.2. The mixture was centrifuged vent hypoglycemia. DMS as compared to DD group.. Ariana Tunis. Antioxidant enzymes and glutathione assays in plasma and kidney 2. Tuni.5-dithiobis-2 nitro benzoic acid) was added to com- cause alloxan is able to produce fatal hypoglycemia as a result of massive pancreatic pounds containing sulfhydryl groups.4.4.6. Catalase (CAT) activity was assayed by the method of (CD). Measurement of malonaldialdehyde (MDA) in tissues Concentrations of MDA in tissues.8). Japan). 24 h before sacrifice day. An amount of 0. the LDL-C ¼ Total cholesterol  ðTriglycerides=5 þ HDL-CÞ. Ellman (1959) and modified by Jollow et al.5. One milliliter solution containing Flax (Linum usitatissimum L. using commercial Kidneys removed from the control and tested rats were cleaned and fixed in 10% reagent kits purchased from Biomaghreb (Ref. buffered formalin solution.4. this is the first biochemical inves- tigation undertaken to explore in alloxan diabetic animals. at 1600g for 15 min. Blood samples were collected from the rats’ tail veins which were fasted overnight to obtain baseline blood glu- cose levels. All data were expressed as means ± SD. Subsequently. ware for Windows (SAS Institute. rats of both control and treated groups were injected 2. in allox- an diabetic animals. Enzymatic reaction was initiated by adding an aliquot of 20 ll of the seeds mixture supplemented at 33% (DMS). CAT activity was calculated in terms of nmol H2O2 consumed/ 2. The digestibility of macronutrients tary evaporator (Heidolph. Plasma insulin was determined using rat insulin enzyme-linked immunosor- bent assay (ELISA) kit Ref. DMS and CD and DMS and DD groups was performed with one-way ANOVA fol- lowed by student t-test. Estimation of insulin concentration considered significant if p 6 0. Body weight gain. 20112. The present study was conducted in three groups of six rats each: control rats 2. The rats received for the next 24 h. Tunisia). Statistical analysis intraperitoneally with glucose (2 g kg1 bw).67% thiobarbituric acid (TBA) and 0. To our knowledge. Table 1. Lipid perox- ga 3 and Omega 6 was prepared. Supernatant (500 ll) were taken and added to Ellman’s re- centration was about 200–300 mg dl1 were taken for the experimental tests. (1974) based on the development of (2002).3. A 500 ll of tissue homogenate in phosphate insulin release. Essex. control and treated rats. Tunisia) and tap water given ad libitum during Units of SOD activity were expressed as the amount of enzyme required to inhibit 30 days of experimental period.5 M in a med- ium containing 100 mM phosphate buffer. Catalase activity (CAT).4. with pellated food (Industrial razolium (NBT).6. Materials and methods mined spectrophotometrically according to Draper and Hadley (1990). diabetic rats (DD) and diabetic rats fed diet containing Flax and Pumpkin Aebi (1984). was mixed with 1 ml of trichloroacetic acid solution and centrifuged at 2500g for 10 min. After 2 weeks of acclimatization. The absorbance was measured at 412 nm after 10 min. Table 1 summarizes body weights. France).1.3-tetraethoxypropane as standard.4. 120 mg kg1 BW of alloxan monohydrate freshly 2. 532 nm using a spectrophotometer (Jenway UV-6305. CA). Total GSH content was expressed as mg/ml in plasma and as mg/mg of protein in kidney. The animals were cared in accordance with the the reduction of NBT by 50% and the activity was expressed as units per mg of principles of the Guide for the Care and Use of Experimental Animals (Council of protein. . 20151. The seeds mixture of Flax and Pumpkin rich in Ome..1.1. according to Mansour et al. Blood was collected from the rats’ tail vein at interval of 30 min up to 2 h for glucose estimation using a glucometer (Esprit The data were analyzed using the statistical package program Stat view 5 Soft- 2. Glutathione levels (GSH). GSH in tissues was determined by the method of prepared in normal saline. Absorbance of TBA-MDA complex was determined at and stored at 4 °C prior to use.4.) seeds were pur. were obtained from the Central ture contained 50 mM of tissue homogenates in potassium phosphate buffer (pH Pharmacy of Tunisia (SIPHAT. Experimental design 2. 0. 3. Makni et al. digestibility and feed conversion efficiency (FCE) 20143. Plasma lipid parame- ters such as total cholesterol (TC). and 20091) purchased from Biomagreb (Ariana. 13 mM L-methionine. a yellow color when DTNB (5. Laborota 4010 digital. They were maintained under standard lab- 7. Biochemical assays 2. seeds according to the method of Folch et al.) and Pumpkin (Cucurbita pepo L. All sections were evaluated for the to a glucose challenge (Matteucci and Giampietro. A significant de- Plasma lipids were extracted with chloroform/methanol mixture (2v/1v) crease of body weight gain was observed in DD rats. The reaction mix- Male Wistar rats (190–210 g) aged 3 months. The results were 2.

Significant increases in plasma TC (137%) and TG (85%) lev. Significant differences between the DMS and DD groups: +p < 0. DD and DMS experimental rats. HDL-C. 200 150 Table 2 100 Plasma levels of creatinine.98 186. els were observed in DD group.42 85. a Values are given as means ± standard deviation (mean of six determinations). DD and DMS groups. digestibility.00 17. Values are given as means of six determinations. Glucose tolerance test (GTT) in control and diabetic rats.61*** 100. increased. HTR ratio and AI are represented in 100 Table 3.85 ± 7. The HTR ratio significantly Flax and Pumpkin seeds mixture significantly increased the tol. feed intake.4.89 3. 400 Blood glucose level (mg/dl) 350 3. Plasma insulin levels in CD. tively. . Effect of seeds mixture supplemented to diet on plasma lipid 300 parameters 250 While the DD group recorded an increase in plasma lipids by 200 108%. means ± standard deviation [mean of six determinations]. 50 0 400 0 30 60 90 120 Time (min) glucose 2g/l Blood Glucose Levels (mg/dl) 350 300 CD DD DMS 250 Fig. (CD) (Fig. The administration of Flax and Pumpkin seeds mix- ture to rats with hyperglycemia resulted in the significant decrease of plasma glucose level in comparison to the result obtained from the DD group. Blood glucose and glucose tolerance test (GTT) Fig. levels of urea.95+++ Days Uric acid (lmol/l) 302.09 ± 2.13 ± 2.18 Significant differences between the DD and CD groups: *p < 0. Flax and Pumpkin seeds mixture supple. Fig.2 0 CD DD DMS 3.2 Parameters and treatments CD DD DMS 1 Body weight gaina (g) 99. LDL/HDL ratio and AI also signifi- parison to the CD group. 3.05. average feces weights. compared to the CD group.6 ± 7.8 FCE ratio 00. 1). compared to those of DD group. Administration of Flax and Pumpkin seeds mixture to diabetic rats significantly reversed these changes to near normal values. and feed conversion +++ efficiency (FCE) in rats fed diets for 4 weeks. LDL-C levels.78+ CD Group DD Group DMS Group Values are given as means ± standard deviation (mean of six determinations). TC. 0.20 00.2 ± 11.34 350. cantly increased in the plasma of the last group. 0.2 ± 5. Blood glucose (mg/dl) levels of CD. DD and DMS groups.24 0. Estimation of urea. M. creatinine and uric acid were sig. A In diabetic rats.55 ± 10. Values are given as means ± The plasma glucose concentration in the diabetic group (DD) standard deviation [Mean of six determinations]. noticed 120 min after glucose injection.54+ *** 0.001. 1. Significant differences between significantly increased in comparison to the normoglycemic group the DD and CD groups ***p < 0.76 ± 6. Values are given as Significant differences between the DD and CD groups: *p < 0. plasma lipids in the DMS group 150 decreased by 21% compared to the DD group (Table 3).43 22.001.66 ± 1. Significant differences between the DMS and DD groups +++p < 0.55 83. uric acid and creatinine MDA levels in plasma and kidney are presented in Table 4.09 02. 3).001. 2) of DD rats decreased by 42% in com. The maximum glucose tolerance was DMS group as compared to those of DD group.001.5.12 13.86+++ Urea (mmol/l) 10. In DMS group mented to the diet of DMS group increased plasma insulin plasma TC and TG levels were decreased by 47% and 47%. (102%) was observed in DD group compared to those of control trols (Table 2). TG. / Food and Chemical Toxicology 48 (2010) 2407–2412 2409 Table 1 1.00* 87. significant increase in MDA levels in plasma (132%) and in kidney nificantly higher in plasma (29%.05 and +++p < 0. Plasma insulin level (Fig. 50 Parameters and CD DD DMS 0 treatments J+0 J+2 J+7 J+14 J+21 J+30 Creatinine (lmol/l) 112.34 0. while LDL/HDL ratio and AI significantly decreased in erance for glucose (Fig.00 72.76*** 297.4 Body weight gain. Plasma insulin levels (ng/ml) 1.2.3. urea and uric acid of CD. 66% and 16%) than those of con.00 Average feed intake (g/d) 20. Makni et al. respec- concentration by 63% in comparison to the DD group.26 Average feces weight (g/d) 03.83* 10. 2.6 Apparent digestibility (%) 84.05.05 and ***p < 0. Lipid peroxidation in plasma and kidney homogenates 3. 3.4 Significant differences between the DMS and DD groups: +p < 0.

24 ± 0. CAT and SOD activities in plasma (42%. The need for more appropriate therapy increases LDL-cholesterol (g/l) 0. group CD.80 ± 0.55 ± 1. CAT) in plasma and kidney of CD.94 ± 0.001.71 ± 0. Discussion Total plasma cholesterol 0.66++ 4. 3.66 ± 0. AI = (TC  HDL)/HDL. .26 ± 1.57+ Kidney 14.11 ± 0.20*** 0.11+++ Values are given as means ± standard deviation (mean of six determinations). Alloxan treatment elicited Table 4 MDA. a MDA = nmol/ml in plasma and nmol/100 mg in liver. jury of tubular and glomeruli (Fig. b GSH = mg/ml in plasma and mg/mg protein in liver.07 1. Parameters and treatments CD DD DMS MDAa Plasma 3.38 20.30 ± 1.7.26 6.91 4.53 ± 2.61 ± 1.40 ± 0.41** 5.28 ± 2. and +++ p < 0. 53% and 35%). as compared to those of CD group. 27% and 60%) and in kidney (34%. as compared to those of DD group.99 ± 3. Thus offer a natural key to unlock a diabetolog- LDL/HDL ratio 0.18+++ (Bailey and Day.65 ± 2. DD (B) and DMS (C) groups c SOD = units/mg protein. Diet supplemented with seeds mixture in DMS group.19** 5.6.77 ± 0.05 1.92*** 5. 3.05.10*** 1.45 8. DD and DMS groups. The present results suggest that Flax and Pumpkin seeds mix- HTR (%) = HDL-C/TC ratio.50 ± 1.33 ± 0. glucose level in diabetic rats is an important basal parameter for Significant differences between the DMS and DD groups: +p < 0.06* 0. Parameters and treatments CD DD DMS Total plasma lipid (mg/ml) 9. Glomerular space reduction. DD and DMS rats.001.01.85 ± 0.20+++ ist’s pharmacy for the future. Diet supplemented with seeds mixture induced a signif- icant decrease of MDA levels in plasma (28%) and in kidney (34%) compared to DD group.32 ± 5.24 ± 0.24 ± 0. respectively. Significant differences between the DMS and DD groups: +p < 0.92 ± 0.71** 18.13*** 0. 400).21*** 1.65*** 36. improved GSH levels.05. / Food and Chemical Toxicology 48 (2010) 2407–2412 Table 3 significant morphological changes in renal DD rats with severe in- Plasma lipid profile in CD. CAT and SOD activities was observed in plasma (57%.07 ± 1.95*** 5. In DD group.10 ± 0. **p < 0.05+++ (g/l) bolic disease.16 ± 0.25+++ SODc Plasma 15.31+++ by recent methods.18 5. respectively. Significant differences between the DD and CD groups: *p < 0.55 ± 3.03 ± 2.04+ betic plants and herbs are known in traditional medicine but their HTR (%) 44.42 ± 0.28 ± 0.93 ± 4.04 4. and protector effects in alloxan-induced diabetic rats.45*** 16.37 9. Fig.64++ CATd Plasma 6.001. Antioxidant enzyme activities and glutathione levels in plasma and kidney GSH levels and antioxidant enzyme activities (CAT and SOD) in the plasma and kidney of control and tested groups are shown in Table 4. ++p < 0.61 13.001. 4A–C).21*** 14.67 ± 0.49 ± 1.30 ± 0. ++p < 0.06*** 14. 1989). ?: Necrosis.76++ GSHb Plasma 8. and +++ p < 0.22 ± 0.39 4.67 ± 0.01. A large number of hypoglycemic/antidia- HDL-cholesterol (g/l) 0.60 ± 0.18+++ introduction into modern therapy waits pharmacological testing Atherogenic index (AI) 1. Arrows indicate: : Tubular dilatation. Significant differences between the DD and CD groups: *p < 0. hypolipidemic and nephro- Values are given as means ± standard deviation (mean of six determinations).32 ± 0.08 1. (hematoxylin and Eosin.12 ± 0. 4.07+++ (TC) (g/l) Diabetes mellitus is possibly the world’s largest growing meta- Plasma triacylglycerol (TG) 0. and ***p < 0. a significant decrease of GSH levels.02 0.01.2410 M.66+++ Kidney 18. 34% and 25%). Kidney histological sections of CD (A).02 ± 0.12*** 0. Light microscopy study of kidney tissue The kidney histological examination of control and DMS groups showed normal cells’ architecture.01.29 9. ture exhibit significant hypoglycemic.45++ Kidney 4.70 ± 0. glutathione levels (GSH) and enzymes activities (SOD. Makni et al.05.05.23 3. **p < 0.53 ± 3.24 ± 0.70 ± 0.40+ Kidney 6.67 ± 1.37 15. : d CAT = lmol H2O2 degraded/min/mg protein. Fasting blood ***p < 0.21 ± 0.65*** 6. 11% and 130%) and kidney (44%.35 ± 1.59 ± 0.

2005). E. R. 1997). M... tected b-cells of the islets of Langerhans (Suba et al. M. N. the decrease of body weight gain in diabetic rats supports these findings. prove hyperglycemia and prevent diabetic complications. health. The significant decrease The biochemical parameters were correlated with the renal his- in the levels of fasting blood glucose in the last group might be ex. Hadley. Anwar and Physiology 135. H.. Omogbai. E. T. Role of oxidative stress in diabetic complications damage. Indian Journal of Experimental Biology ity is through generation of free radicals.. Processing Department of Sidi Bouzid Institute (ISET) Tunisia and down of lipids and mobilization of free fatty acids from the periph. 2007). 2007. pp. Day. Estimation of the concentration animals ensured the regeneration or the protection of pancreatic of low density lipoprotein cholesterol in plasma without use of the preparative b-cells.J. (Baynes and Thorpe.. Analytical Biochemistry 44. M. Fredrickson. cholesterolemic rats (Makni et al. The administra- Alloxan-induced diabetes.. induced by the degradation of betic rats improved the histological alterations induced by structural proteins (Rajkumar et al.. J. 1988. Methods in Enzymology 105. seeds mixture may be due to its protective role in quenching free Erah. we have demonstrated that Flax and Pumpkin peroxidation. Thus seeds mixture has a significant Ahmed. Levy. 27(48). Thorpe. (a new perspective to an old paradigm). D. potentiating of insulin secretion from pro. Malondialdehyde determination as index of lipid previous study. So. atic b-cells. I. Osmde.H. Momordica charantia and and Pumpkin seeds mixture supplemented in the diet of DMS Eugenia jambolana (Sharma et al.O.. 1986)... Vilstrup. Clinical Chemistry 226. Hypoglycemic effect of Ocimum sanctum leaf extract in normal and streptozotocin diabetic rats. of Histopathology Laboratory of CHU Habib Bourguiba. In our study. 70–77. Conflict of Interest Diabetes mellitus is usually associated with an increase in plas- ma lipids levels. Ahmed et al.. 1988).. demonstrated both in vivo and in vitro (Grandy et al. 2000.. Hypoglycemic effect of the extract of radicals (which destroy pancreatic cells) to ensure beneficial role Solenostemon monostachysleaves. Schneider. W.. 1981. S. The concentrations of the metabolites increase in diabetes in rats. reducing the fasting blood glucose level. Cummings. 2004). Nutrition Clinique et Métabolisme 20. New York. In our Draper. Comparative Biochemistry acylglycerol and cholesterol levels (Madinov et al. vol. Biochemistry 261. Stanley. 1998. diabetic rats: effects of garlic oil and melatonin. The present work was supported by the DGRST grants (Appui à 30 days normalized lipid profile in diabetic animals. lipid peroxidation.. and is considered Aebi..L. C. 167–171. resistance? Indian Journal Medical Research 125. (Davidson. In fact.M. Tunisia. Beneficial effects and mechanism of action of Momordica charantia juice in the treatment of impact to improve the imbalance in lipoprotein metabolism. while significant decrease in these parameters was ob.P. blood during renal diseases or renal damage associated with Al-Shamaony. Enhanced plasma insulin levels in the DMS 226 (1). A simple method for the isolation and insulin levels of diabetic treated rats (DMS group) compared to purification of total lipids from animal tissues. Current Atherosclerosis 3. M.. W. (Oi et al.. there was an elevation in Diabetes Care 12. to be a cardio protective lipid. 1976).L. 2003.. TG and LDL but also enhanced the HDL-cholesterol which is known to play an important role in the References transport of cholesterol from peripheral cells to the liver by a pathway termed ‘‘reverse cholesterol transport”. Fridovich. L.T. S.. Duckworth. R.. 1999.. Makni et al. It has shown that Flax medicinal plants like Terminalia chebula. C. 891–893. uric acid. 499–502. So optimum chemical composition of body fluids is Almdal. cals and to subsequent lipid peroxidation induced by alloxan. Tuni- eral stores. assay applicable to acrylamide gel. In the present study. seeds mixture possesses free radical scavenging activity in hyper.S.K. 1972. we revealed that alloxan caused a signif- plained by the stimulation of the residual pancreatic mechanism. Friedewald. Beauchamp. Diabetes 48... and hydroperoxides accumulated in kidney could cause cytotoxicity probably by increasing peripheral utilization of glucose (Erah associated with membrane phospholipids peroxidation. 1959. Superoxide dismutase: improved assays and an served in the animals of DMS group.I. due probably to the generation of reactive radi- destroyed by alloxan..E.. species have been implicated in diabetes and its complications Davidson.M. Administration of Flax and Pumpkin seeds mixture for sia. 355–372. and increased tri. Diabetologia 31. 63–70. icant damage in renal structure showing marked glomeruli and regeneration or protection of pancreatic cells that were partially tubular damages.M. Al-khazrajoi.. 1996). 1994). 1984. It has been suggested that a variety Wiley. G.R. Seed mixtures la Recherche Universitaire de Base ARUB 99/UR/08-73)... C.. characterized by a severe loss in tion of Flax and Pumpkin seeds mixture through the diet of dia- body weight (Odetola et al. I. G. 121–126. This effect has been reported by others with some other ultracentrifuge. H. J. Chattopadhyay. Singh. Journal of Biological Chemistry the DD animals. uric acid and creatinine in the DD group indicating renal Baynes. Diabetes Mellitus Diagnosis and Treatment. Hypoglycemic effect of uncontrolled diabetes mellitus (Almdal and Vilstrup.. Day. Official Journal of the European Communities (JO 86/609/CEE) L358.. 276–287. Molecular and Cellular Kidneys remove metabolic wastes such as urea. 1981. S. Antidiabetic activity of Ellman.. Alloxan Blandeau.. Hyperglycemia and cardiovascular disease. which has been 31. not only lowered the TC. Effect of a valuable extract on some blood parameters in diabetic animals. Antioxidants: do they have a role in the treatment of insulin in survival of pancreatic b-cells. 2006)..C. 1986. 1993.. Meki. creat.K. Council Instructions about the Protection Papaccio et al. A decrease of serum lipid concentration through drug therapy or dietary measures Acknowledgements seems to decrease the risk of vascular diseases (Rhoads et al.. 1957. Lees. Council of European Communities. G. Journal of Ethnopharmacology 43.. Strict insulin treatment normalizes the organic nitrogen contents and the capacity of urea–N synthesis in experimental maintained. 1971. 497–509. TG and TC levels in alloxan diabetic rats The authors thank the skillful technical assistance of the Food observed in the present study may be a result of increased break. E. One of the intracellular phenomenons for its cytotoxic. which could be attributed to its antiradical/antioxidant classic symptoms of diabetes including polydipsia and polyuria activities. Bailey. Murali group causes the hypoglycemic and antihyperglycemic effects by et al. / Food and Chemical Toxicology 48 (2010) 2407–2412 2411 monitoring diabetes (Rajkumar et al.T. 2008).B. 1996. 553–564. C.. 1994. 1999). Evans.. urea.. Flax and Pumpkin seeds mixture caused significant increase in Folch. Free radicals and associated reactive oxygen of Living Animals Used in Scientific Investigations. 1–18. II. pp. for renal cellular damage and necrotic renal cells. 21–27. Tissue sulfhydryl groups.A. Omega-3 fatty acids for mother and child produces hyperglycemia by selective cytotoxic effects on pancre. diabetes was induced by alloxan. and Meki.M. may be due to metabolic disturbance in diabetes reflected in high Anwar. Archives of Biochemistry 82. Methods in Enzymology 86..W. 2003). Al-Shamaony et al. 109–157. A. H.S.R. 114–118.. Oxidative stress in streptozotocin-induced activities of xanthine oxidase. . In the present study. 2004. 421–431. Twaij.. Catalase in vitro.A. H.R. inine and ions... 1990. streptozotocin-induced diabetes mellitus in rat. 68–72.. J. Journal of West African Pharmacy 10. 1982. of antioxidants that scavenge reactive oxygen species might im. Sharma. 1991). Increase in lipids.H.. J. A. Adeghate. 2003. the basis et al. 539–547. 2006. M. 1–9... British Journal of Nutrition 80. 1989. 2004.. P. 381–391. 2001.. tological studies. 203–208. Traditional plant treatments for diabetes mellitus: pharmaceutical foods. were associated with alloxan. Traditional plant medicines as treatments for diabetes. Sfax. This Artemisia herba alba.. the risk factor for coronary heart diseases The authors declare that there are no conflicts of interest.

L. Gulbrandse. 147–152.F.. Babady. R. 2003. 2005. catecholamine. Grigg... O. Adekunle. N. hypoglycaemic herbs on the activity of phase I and II drug-metabolizing 24–38..S. 61–69. Ugochukwu. C. term effects of Terminalia chebula Retz.A.M.. G. P. enzymes in alloxan-induced diabetic rats. Bhaskara. 2003. Terapevticheski arkhiv 72.. glucagon.A. Antioxidant effects of Gongronema latifolium Murali. insulin.. 3714–3720.T. Indian Journal of Medical Research 116.H.. Akinloye. Current Medicinal Chemistry 13.A. Attenuation of hyperglycemia and Zeghal. C. 641–646. Food and Chemical Toxicology 46. Pharmacology and Physiology 33. Ghosh. D. 55–58.. P. India. Changes in plasma glucose. Panikar. diabetes mellitus. tissue glycogen content and in vitro release of insulin in Ugochukwu.. S. 163–172.B.. 2003. Kagen.. alloxan-induced diabetic rabbits.. Journal of Ethnopharmacology 85. D. rats. Ethnopharmacology 115..A. M.. P. oxidative stress and Sheweita.. Murthy.. Ghosh. Markov. Acetyl-homocysteine-thiolactoneinduced liver necrosis: protective role of glutathione and evidence for 3. D. / Food and Chemical Toxicology 48 (2010) 2407–2412 Grandy. H.R... N. V. Oxidative stress and gene expression Pradeepa. Prabhu. H.. O.. Dev.R.. Plant foods in the management of diabetes mellitus: Karunanayake. S. P.R.. H. Uttam Kumar. C. 1–4. 1982. L. S.H.S. Nasir. New Delhi.. Murthy. T. Gassett. D.. Gillete... Indian Journal of Experimental Biology 31. Indian Journal of Experimental diabetes mellitus. N.H. Journal of Bioscience 28.E. Journal of Antidiabetic potential of Barleria lupulina extract in rats.. and glycogen contents in tissues during development of alloxan Ha.G.L.. W. 1986. Kajinuma.. Makni. Rajkumar.A. Limaye. E.A. Diabetes Research and Clinical Practice 45.. Mankil. Govindarajulu. 2001. Makni et al.. of streptozotocin. 201–206. K. M. N. Mohan. S. Egunjobi...A.. Komori.. 1993.. A..K. Mitchell. Jollow.. 2008.C. 808–812.. S.... K. The New 1203–1218.. Paulose. Singh. J. Srinivasan. Chandra. D.K. S. L. agreed diagnostic procedures in experimental diabetes research. Pharmacology 109.. Search of novel hypoglycemic agents vegetables as potential hypoglycemic agents. P.L. publique de Montréal.... A... Gokkusu. N... In: Sharma. Sayeda. Murugesan. Ayoola.A. 1999.. The effect of streptozotocin-induced diabetic rats. M. Rajkumar. S. 1974. Sacks. K. Matteucci.. 1991. 70–75.. Cobourne. Hyun.S. Boudawara. H. Long in hepatocytes of rat models of non-insulin dependent diabetes mellitus. 2003.. T. K. Makni. Ademoglu.I... 1172–1176. Prévention en pratique médicale. Clinical and Experimental Chimica Acta 315. Pal. A.H.. H. S. rats. H. Screening for diabetes: is it warranted? Clinica (JACQ) extract in alloxan-induced diabetic rats. 1997. Mansour. Nahrung 41. V. The changing of the diabetes epidemic implications of antioxidant enzymes in the renal cortex of streptozotocin induced diabetic for India. Effect of leaf extract of Aegle Complications. stress and protein kinase C.. Mansour.K. Moonsoo. Sang. Raghuram. 650–658. hepatocytes of diabetic rats. V.. Buse. 2004.. J.K. Fitoterapia 75... marmelose in diabetic rats. Tandon. .. 68–74.. Maritim. M.. Balasubramanian.. Rhoads. Sanders. Kim. 2002..E. Increased Madinov.. Hypolipidemic and hepatoprotective effects of flax and hyperlipidemia in streptozotocin-induced diabetic rats by aqueous extract of pumpkin seed mixture rich in x-3 and x-6 fatty acids in hypercholesterolemic seed of Tamarindus indica.4 increase of superoxide dismutase counteracts the effect of subdiabetogenic doses bromobenzeneoxide as the hepatotoxic intermediate. Giampietro.H. Rao R. 121–132. from medicinal plants. Crouch. Newairy. A. 612–618. Eun.. Bromobenzene induced Papaccio.. A review.... N.. Bulletin de la Direction de santé Oi. Pisanti. M.. El.I.. 1–5. Palanduz. G. Watkins.G.. (Ed. An Update. K. 293–298. Anand... Molecular and Cellular Biochemistry 243. 1976.I.. K. A.. H..2412 M. Proposal open for discussion: defining Suba.. K. J. Pharmacology 11. England Journal of Medicine 294. R.ca. Yoon-Ho. Frascatore. 221–228. J. on hyperglycemia and associated Fitoterapia 73. 309–318. C.O. Zampaglione. antidiabetic and antihyperlipidaemic effect of fermented Parkia biglobosa Greenberg. J. Tennekoon. P. 2002. http://www. P. R. 1997. M. 2002.K.qc. A. 2007. Main causes of hyperuricemia in degradation of dermal collagen in diabetic rats. Youssef. Protective role of superoxide dismutase Odetola.. Fetoui. Diabetes Mellitus and its Ponnachan. 131–139. K. Biological and Pharmaceutical Bulletin 28.A. R. Possible against diabetogenic drugs. 2002. C. Mandal. R.V. 2002.K.. The Journal of Clinical Investigation 70. S. Garoui. Toxicology 174.. 345–347. N. M.. Patel. Gargouri.. Babady. 1993.. E. R. 1081–1083. A. Effect of some antioxidants..S.. Sheweita. hyperlipidemia. Serum lipoproteins and coronary Antidiabetic agents from medicinal plants.K. S. Biochemical study on Sharma. MacMillan India Ltd. M.. Srinivasan. N. K. Biochemical and Molecular Medicine 62. I. 2006. 2008.B. Youssef. Plasma antioxidants and type 2 147–151. S.).B.V.J. Jaber.. 2000. E.C. 2004. Diabetes 35 (4)..M.. A. 151–169.R. artery disease in a population study of Hawaiian Japanese men. G. N. Experimental and Clinical Endocrinology Gongronema latifolium extracts on serum lipid profile and oxidative stress in and Diabetes 115. K. Pathogenesis of diabetic nephropathy: the role of oxidative diabetes mellitus in rats.C. 2006. F. Journal of Biochemistry and Molecular Toxicology 17. Sivakami. 470–474.. Y... Toxicology hypolipidemic effect of ethanolic extract of seeds of Eugenia jambolana in 170. Balabolkin. Hypoglycemic and the effects of some Egyptian herbs in alloxan-induced diabetics rats..santepub-mtl.. Diabetes. Biology 29.A.