Effect of flax seeds, pumpkin seeds and sesame seeds on liver enzymes and kidney functions of rats suffering from osteoporosis
Atiat Mohamed EL-BAHY(1), Sanaa El-Bendary(2), Ashraf A. A. El-Megeid (1), Ekbal Mahmoud(2), Yasmeen Fahmy Shoib(3), Thanaa Mahmoud Hashem(3).
(1): Faculty of home economic, Nutrition and Food Science Department Helwan University,
(2): Nutrition and Food Science, Department of Home Economics, Faculty of Education,
Ain Shams University,
(3): Faculty of specific education, Fayoum University.
ABSTRACT
This study was carried out to investigate the effect of two levels from flax seeds, pumpkin seeds, sesame seeds and combination on liver enzymes and kidney functions of rats suffering from osteoporosis. Sixty female rats (Sprague dawley strain) weighting an average of (200±10g) used in this study. The rats were divided into two main groups. The first main group (6 rats) was fed on basal diet as a (control negative group).The second main group (54 rats) fed on diet containing 100 mg (prednislone acetate) as source of Glucocorticoid GC/kg diet to induce osteoporosis for two weeks. Then the second main group divided into nine subgroups (n=6 each) as follows: subgroup (1) fed on prednislone acetate diet as a (control positive group), subgroups (2 and 3) fed on prednisone acetate diets containing (5% and 10%) flax seeds. Subgroups (4 and 5) fed on prednisone acetate diet containing (5% and 10%) pumpkin seeds. subgroups (6 and 7) fed on prednisone acetate diets containing (5% and 10%) sesame seeds. subgroups (8 and 9) fed on prednisone acetate diet containing (5% and 10%) combination of flax seeds, pumpkin seeds and sesame seeds. The results revealed that, food intake, body weight gain%, kidney functions (uric acid, urea nitrogen and creatinine) and liver enzymes (AST, ALT and ALP) of all treated groups improved than that of the positive control group, especially the osteoporosis group fed on diet containing 10% combination of (flax seeds, pumpkin seeds and sesame seeds). The pathological examination of bone confirmed this results.
Key words: Flax seeds, Pumpkin seeds, Sesame seeds, Kidney function (Uric acid, Urea nitrogen and Creatinine) and liver function (AST, ALT and ALP).
INTRODUCTION:
Osteoporosis is a disease of bone that leads to an increased risk of fracture. In osteoporosis the (BMD) is reduced, bone micro architecture was disrupted. It is a bone condition defined by low bone mass, increased fragility, decreased bone quality and an increased fracture risk (WHO, 1994). Some studies reported that flax seeds, pumpkin seeds and sesame seeds help to improve kidney functions and liver enzymes of rats suffering from osteoporosis. Flax plant contains substances which promote good health. Flaxseed and flaxseed oil are rich in alpha-linolenic acid (ALA), an essential fatty acid that appears to be beneficial for heart disease. ALA belongs to a group of substances called omega-3 fatty acids (Urology 2001). Flax seed is high in most of the B vitamins, magnesium, and manganese. Also flax seeds contain Omega-3 fatty acids is a key force against inflammation in our bodies. Flax seed is high in phytochemicals, including many antioxidants. It is perhaps our best source of lignans, that lignans may promote fertility, reduce peri- menopausal symptoms, and possibly help prevent breast cancer. In addition, lignans may help prevent Type 2 diabetes (Laura Dolson, 2008). Flax seed is rich in omega 3 fatty acid that is responsible for its ability to reduce bad cholesterol, triglycerides and blood pressure. It also battles inflammatory reactions in the body (Evelyn De Matias, 2010).
Pumpkin seeds and sesame seeds are improving kidney functions and liver enzymes of rats suffering from osteoporosis. Pumpkin seeds contain omega-3 fats and zinc, pumpkin seeds protect men's bone andante- inflammatory benefits in arthritis but do not increase the level of damaged fats (lipid peroxides). Pumpkin seeds are a very good source of the minerals magnesium, manganese and phosphorus, and a good source of iron, copper, protein and vitamin K(Ensminger AH, et al 1986).
Pumpkin seed is rich sources of polyunsaturated fatty acid (PUFAs) and mostly lipophilic antioxidants may help to prevent osteoporosis associated with estrogens (Boulbaroud, S et al., 2008). Sesame seeds are a very good source of copper and a good source of magnesium and calcium, sesame seeds contains calcium which helps to prevent colon cancer and osteoporosis. Also it contains zinc which protect of osteoporosis (Ensminger AH, et al 1986).
Sesame seeds are a source of helpful biologically active components found in plant foods, such as phytochemicals and it is a functional food that could be used in the prevention, controlling to illnesses such as cancer, oxidative stress, cardiovascular disease and osteoporosis (Sacco, et al., 2008). The aim of this study was to investigate the effect (flax, pumpkin and sesame) seeds on liver enzymes and kidney functions of rats suffering from osteoporosis.
MATERIAL AND METHODS Materials:
- Casein, vitamins, minerals, cellulose, choline chloride were purchased from El- Gomhoreya company, Cairo Egypt.
- Sixty female rats (Sprague Dawley Strain) were obtained from Helwan farm.
- Prednisone acetate was purchase from some pharmacy in Cairo, Egypt.
- Flax seeds, pumpkin seeds and sesame seeds were obtained from local marked (Cairo-Egypt).
Experimental animals:
Sixty Female rats( Sprague Dawley strain) weighting an average of (200 ±10g.) was housed in well aerated cages under hygienic condition and fed on basal control diet for one week for adaptation , the basal diet prepared according to (Reeves et al, 1993), salt mixture which used in this study according to (Hegsted et al., 1941) and vitamin mixture prepared according to (AOAC., 1975).After this period, the rats were divided into two main groups , as a following: The first main group (6 rats), was fed on basal diet, as a control negative group. The second main group (54 rats) fed on basal diet containing 100 mg (prednisone acetate) as source of glucocorticoid GC/ kg diet to induce osteoporosis for two weeks to induce osteoporosis according to (Liao et al., 2003). After this time, the second main group divided into nine subgroups (6 rats each):
subgroup (1): fed on prednisone acetate diet, as a control positive group.
Subgroups (2 and 3) fed on prednisone acetate diets containing 5% and 10 flax seeds, respectively. Subgroups (4 and 5) fed on prednisone acetate diets containing 5% and10% pumpkin seeds, respectively.
Subgroups (6 and 7) fed on prednisone acetate diets containing (5% and 10%) sesame seeds, respectively. Subgroups (8 and 9) fed on prednisone
acetate diets diets containing 5% and 10% combination of (flax seeds, pumpkin seeds and sesame seeds), respectively.
During the experimental period (28 days), the diets consumed and body weights were recorded twice weekly according to (Chapman et al., 1959). At the end of the experimental, the animals were fasted overnight, then the rats were anaesthetized and sacrificed and the serum was separated to estimate some biochemical parameters, i.e. serum aspartate amino transferase (AST) and alanine amino transferase (ALT) by (Ritman and Frankel, 1957), alkaline phosphatase (ALP)by (Bergmeyer and Brent, 1974) uric acid (Fossati et al., 1980), urea nitrogen (Patton and Crouch 1977), creatinine by (Bohmer, 1971).
The liver in each group was examined histopathologicaly, according to (Sheehan and Hrapchak 1980). The data obtained was analyzed statistically for standard deviation and one-way ANOVA test (Armitage and Berry, 1987).
RESULTS
Effect of flaxsseds,pumpkin seeds and sesame seeds on food intake and body weight gain% of rats suffering from osteoporosis.
Table (1) showed the effect of low and high levels from flaxseeds, pumpkin seeds, sesame seeds and their combination on food intake and body weight gain% of rats suffering from osteoporosis. Food intake of osteoporotic group decreased than that of healthy rats (the percent decrease in food intake was 24.326%). The mean value of food intake in all treated groups increased, as compared to the positive control group.
Osteoporotic groups which treated with 10% flaxseeds or 10%
combination of (pumpkin seeds, sesame seeds and flaxseeds) recorded the highest increase in the mean value of food intake, as compared to other treated groups.
Data in the same table showed significant decrease p< 0.05 in body weight gain % (BWG %) in osteoporotic group (fed on Prednisone acetate diet), as compared to healthy rats (fed on basal diet). All treated groups recorded significant increase in BWG%, except group of rats treated with prednisone acetate diet supplemented with 5% pumpkin seeds, as compared to the positive control group. Data in this table revealed that, non-significant differences in BWG % was observed between all osteoporotic groups which treated with low level (5%) from pumpkin seeds, sesame seeds and flaxseeds. The same trend was observed whine between groups which treated with the high level (10%).
The best results in body weight gain percent were showed in the treated group with the high level from the combination of (pumpkin seeds, sesame seeds and flaxseeds), because the BWG% in this group showed significant increase, as compared to other treated groups.
Table (1): Effect of flax seeds, pumpkin seeds and sesame seeds on food intake and body weight gain % of rats suffering from osteoporosis.
Parameters groups
Mean of Food intake (g/day/rat)
Body weight gain
%
Control (-ve) 15.432 31.078 ± 2.293 a
Control (+ve) 11.678 -32.119± 5.595 e
5% pumpkin seeds (PS). 12.439 -26.808± 2.020 d e 10% pumpkin seeds (PS). 12.985 -20.958± 3.785 c d 5% sesame seeds (SS). 12.509 -28.824± 3.134 e 10% sesame seeds (SS). 13.095 -20.407± 8.095 c d 5% flax seeds (FS). 12.773 -26.465± 5.127 d e 10% flax seeds (FS). 13.439 -17.493± 6.261 b c 5% combination of (PS, SS and FS). 12.966 -20.701± 8.923 c d 10 % combination of (PS, SS and FS). 13.553 -12.325± 4.968 b
- Values are expressed as mean ± SD.
- Significant at p<0.05 using one way ANOVA test. - Values which have different letters differ significantly, while those with have similar or partially are not significant.
Effect of flax seeds, pumpkin seeds and sesame seeds on organs weight / body weight % of rats suffering from osteoporosis.
Liver and kidney weight/ body weight % of healthy rats, untreated osteoporotic rats and treated with two levels from flaxseeds, pumpkin seeds, sesame seeds and their combination showed in table (2). The data revealed that, the mean value ± SD of liver and kidney weight/body weight% of osteoporotic rats (control+ve group) increased significantly P<0.05, as compared to healthy rats (control-ve group). Liver and kidney weight /body weight% of all treated groups decreased significantly, as compared to the positive control group. While liver and kidney weight/body weight% of all treated groups showed significant increasing, as compared to control negative (rats fed on basal diet).
Non-significant changes in liver weight / body weight % between treated groups with 10% (flax, pumpkin and sesame seeds) and 5%
combination. The best results in liver and kidney weight/body weight gain% were appeared in osteoporotic group treated with diet containing 10% combination of flaxseeds, pumpkin seeds and sesame seeds.
Table (2): Effect of flax seeds, pumpkin seeds and sesame seeds on liver and kidney weights / body weight % of rats suffering from osteoporosis.
Parameters groups
Organs weights / body weight
%
Liver Kidney
Control (-ve) 3.226de
± 0.138
0.611e
± 0.035
Control (+ve) 5.406a
± 0.182
1.150a
± 0.078
5% pumpkin seeds (PS). 3.702b
± 0.127
0.921b
± 0.031 10% pumpkin seeds (PS). 3.382cd
± 0.123
0.798c
± 0.043
5% sesame seeds (SS). 3.756b
± 0.163
0.879b
± 0.072 10% sesame seeds (SS). 3.361cde
± 0.168
0.791c
± 0.085
5% flax seeds (FS). 3.528c
± 0.166
0.808c
± 0.058
10% flax seeds (FS). 3.324de
± 0.084
0.789c
± 0.063 5% combination of (FS, PS and SS). 3.369cd
± 0.108
0.690d
± 0.051 10% combination of (FS, PS and SS). 3.191e
± 0.045
0.664de
± 0.028
- Values are expressed as mean ± SD.
- Significant at p<0.05 using one way ANOVA test. - Values which have different letters differ significantly, while those with have similar or partially are not significant.
Effect of flax seeds, pumpkin seeds and sesame seeds on kidney functions (uric acid, urea nitrogen and createnine) of rats suffering from osteoporosis.
Data presented in Table (3) indicated that, the value of serum uric acid, urea nitrogen and creatinine (mg/dl) in the osteoporotic group
(control + ve) increased significantly p<0.05, as compared them with the negative control group.
Treating osteoporotic groups with supplemented diet with 5% and 10% (flax seeds, pumpkin seeds, sesame seeds and their combination) decreased the mean values of serum uric acid, urea nitrogen and creatinine, as compared to the positive control group.
The obtained results indicated that, serum uric acid, urea nitrogen and creatinine decreased gradually with increasing the levels of flaxseeds, pumpkin seeds, sesame seeds and their combination.
Treating osteoporotic rats with flaxseeds, pumpkin seeds and sesame seeds individually improved serum uric acid, urea nitrogen and creatinine), while the combination of them showed the best results in these parameters.
Table (3): Effect of flax seeds, pumpkin seeds and sesame seeds on kidney functions of rats suffering from osteoporosis.
mg/dl Parameters
Groups
Creatinine Urea
nitrogen Uric acid
0.508 f
±0.028 31.969 g
±1.406 1.610 g
±0.036 Control (-ve)
1.264a
±0.059 58.987 a
±3.422 2.465 a
±0.071 Control (+ve)
0.912 c
±0.048 46.746 c
±0.985 1.893d
±0.029 5% flax seeds
0.739 e
±0.029 34.771f
±2.253 1.776e
±0.036 10% flax seeds
0.953c
±0.058 49.989 b
±2.005 2.082c
±0.052 5% pumpkin seeds
0.771de
±0.046 38.748 e
±1.476 1.837 de
±0.034 10% pumpkin seeds
1.026 b
±0.064 52.001b
±2.771 2.160 b
±0.081 5% sesame seeds
0.796de
±0.031 40.039 de
±1.683 1.862 d
±0.061 10% sesame seeds
0.815d
±0.062 41.776 d
±2.380 1.834de
±0.043 5% combination of (FS,PS
and SS)
0.665f
±0.057 33.078 fg
±1.864 1.702 f
±0.028 10% combination of (FS,PS
and SS)
- Values are expressed as mean ± SD.
- Significant at p<0.05 using one way ANOVA test. - Values which have different letters differ significantly, while those with have similar or partially are not significant.
Effect of flaxseeds, pumpkin seeds and sesame seeds on liver enzymes of rats suffering from osteoporosis.
Table (4) presented the effect of (flaxseeds, pumpkin seeds, sesame seeds, combination of them) with two levels on AST, ALT and ALP (U/L) of rats suffering from osteoporosis.
Table (4): Effect of flaxseeds, pumpkin seeds and sesame seeds on liver enzymes of rats suffering from osteoporosis.
- Values are expressed as mean ± SD.
- Significant at p<0.05 using one way ANOVA test. - Values which have different letters differ significantly, while those with have similar or partially are not significant.
The mean values of AST, ALT and ALP for control positive group showed a significant increase p<0.05 in AST, ALT and ALP, as compared to the control negative group. All groups which were fed on prednisone acetate diets containing (flaxseeds, pumpkin seeds, sesame seeds and their combination) showed significant decrease in the mean values of AST, ALT and ALP, as compared to the positive control group.
U/L Parameters
Groups AST ALT ALP
84.531g
±3.012 30.005 g
±1.689 76.836h
±2.771 Control (-ve)
178.281a
±5.715 65.559 a
±4.049 152.197 a
±4.819 Control (+ve)
131.991e
±4.169 50.705 c
±3.083 120.971 c
±4.134 5% flax seeds
118.846 f
±4.065 36.979f
±2.620 90.303 f
±4.568 10% flax seeds
152.258c
±3.372 55.436 b
±2.476 134.069 b
±3.551 5% pumpkin seeds
135.991e
±4.247 41.366 e
±1.997 102.218e
±3.455 10% pumpkin seeds
163.082 b
±5.034 58.265 b
±2.114 138.189 b
±3.587 5% sesame seeds
141.255 d
±3.360 45.952 d
±2.483 109.187 d
±3.150 10% sesame seeds
116.303 f
±2.838 42.656 e
±3.171 102.076 e
±5.425 5% combination of (FS,PS and
SS)
103.428 g
±5.788 34.945 f
±1.813 82.169 g
±5.369 10% combination of ( FS,PS and
SS)
The results in this table revealed that, the mean values of AST, ALT and ALP decreased gradually with increasing the levels of flaxseeds, pumpkin seeds, sesame seeds or the combination of them.
The lowest mean values of AST, ALT and ALP recorded for the group fed on prednisone acetate diet containing 10% combination of flaxseeds, pumpkin seeds and sesame seeds , followed by the group fed prednisone acetate diet containing 5% combination of them. On the other hand the highest mean value of AST, ALT and ALP observed in the group treated with 5% sesame seeds, followed by the group which treated with 5% pumpkin seeds, respectively.
Feeding rats which were suffer from osteoporosis on diet containing high levels from combination of (flaxseeds, pumpkin seeds and sesame seeds) recorded the best results in AST,ALT and ALP, this group showed significant decrease in these parameters, as compared to other treated groups.
HistoPathological changes in liver
Photo (1): Liver of rats fed on basal diet (control -) show no histopathological changes of hepatic lobule. (H and E X400).
Photo (2): Liver of rats fed on prednisone acetate diet (control +) showing cytoplasmic vacuolization of hepatocytes and pyknosis of hepatocytic nuclei. (H and E X400).
Photo(3): Liver of rats fed on prednisone acetate diet containing 5% flax seeds showing Kupffer cells activation. (H and E X400).
Photo(4): Liver of rats fed on prednisone acetate diet containing 10% flax seeds showing cytoplasmic vacuolization of hepatocytes. (H and E X400).
Photo (5): Liver of rats fed on prednisone acetate diet containing 5% pumpkin seeds showing cytoplasmic vacuolization of hepatocytes. (H and E X400).
Photo (6): Liver of rats fed on prednisone acetate diet containing 10% pumpkin seeds showing cytoplasmic vacuolization of hepatocytes. (H and E X400).
Photo (7): Liver of rats fed on prednisone acetate diet containing 5% sesame seeds showing Kupffer cell activation and presence of small vacuoles in the cytoplasm of hepatocytes. (H and E X400).
Photo (8): Liver of rats fed on prednisone acetate diet containing 10% sesame seeds showing cytoplasmic vacuolization of hepatocytes. (H and E X400).
Photo (9): Liver of rats fed on prednisone acetate diet containing 5% combination of flax seeds, pumpkin seeds and sesame seeds showing small vacuoles in the cytoplasm of sporadic hepatocytes. (H and E X400).
Photo (10): Liver of rats fed on prednisone acetate diet containing 10%
combination of flax seeds, pumpkin seeds and sesame seeds showing cytoplasmic vacuolization of hepatocytes. (H and E X400).
DISCUSSION:
Our results revealed that, feeding rats on diet containing prednisone acetate to induce osteoporosis decreased food intake and body weight %, while treated rats which were suffer from osteoporosis with 5% and 10% (flaxseeds, pumpkin seeds, sesame seeds and their combination) improved the percent of body weight. In this respect Jane
et al., (2000) reported that administration of glucocorticoids at a dosage of 5 mg/kg/d for 5 days would result in a decrease in both body and diaphragm weights.
Treating rats which were suffering from osteoporosis with 10%
(pumpkin seeds or sesame seeds and/or flaxseeds) also the low and high levels from the combinations showed non-significant different in liver weight, as compared to the negative control group. The Histopathological examination confirmed these results.
There are non-significant different in 10% between treated groups with 5% (pumpkin seeds and sesame seeds), also non-significant differences in kidney weight / body weight % were observed between treated groups with dietary supplemented with 10% flax seeds, 10%
pumpkin seeds, 10% sesame seeds and 5% flax seeds. The best results in kidney weight were appeared in treated group with 10% combination of flax seeds, pumpkin seeds and sesame seeds.
There are significant decrease p<0.05 in levels of kidney fuctions and liver enzymes (uric acid, urea nitrogen, creatinine, AST, ALT and ALP) in groups of rats treated with dietary supplemented with two levels from flax seeds, pumpkin seeds, sesame seeds and their combinations, as compared to un-treated group (osteoporotic group). The highest improvement in kidney functions and liver enzymes recorded for the osteoporotic groups treated with low and high levels from the combination of (flax seeds, pumpkin seeds and sesame seeds).
In this respect, Bhathena et al (2002) investigated the effect of dietary soya protein and flaxseed meal on cholesterol, triglyceride, uric acid, blood urea nitrogen (BUN), creatinine and total protein was studied in two animal models, F344 rats with normal lipid levels and obese SHR/N-cp rats with elevated levels of cholesterol and triglyceride. The rats were fed either 20% casein, 20% soya protein or 20% flaxseed meal.
The authors reported that, flaxseed meal lowered uric acid and BUN.
Omage et al., (1998) reported that, administration of sesame seed oil with the high-fat diet decreased (P<0.01) uric acid concentrations compared with the rats given the high-fat diet only.
Nkosi, et al (2005). studied the effects of pumpkin seed (Cucurbita pepo) protein isolate on the activity levels of lactate dehydrogenase (LD), alanine transaminase [alanine aminotransferase] (ALT), aspartate transaminase [aspartate aminotransferase] (AST) and alkaline phosphatase (ALP) against carbon tetrachloride (CCl4)-induced acute liver injury in low-protein fed rats were investigated. The results were significant improvement in the activity levels of all 4 enzymes investigated.
Uchida et al., (2007) mentioned that there is dietary sesame seed decreases urinary excretion of alpha - and gamma -tocopherol metabolites in rats. The aim of this study was to clarify the effect of dietary sesame seed on alpha -tocopherol metabolism. Urinary excretion of 2, 5, 7, 8- tetramethyl-2(2'-carboxyethyl)-6-hydroxychroman (alpha -CEHC), a alpha -tocopherol metabolite, in rats fed alpha -tocopherol with sesame seed was inhibited (p<0.05) as compared with that in rats fed alpha - tocopherol alone. The inhibition of alpha - and gamma -CEHC excretion by sesame seed was accompanied by elevation (p<0.05) of the alpha - and gamma -tocopherol concentration in the liver. These results suggest that dietary sesame seed inhibits not only gamma -tocopherol metabolism to gamma -CEHC but also alpha -tocopherol metabolism to alpha -CEHC in rats.
Ikeda et al., (2007). Indicated that dietary sesame seed and its lignan increase both ascorbic acid concentration in some tissues and urinary excretion by stimulating biosynthesis in rats. The dietary sesamin and sesame seed elevated ascorbic acid concentrations in the liver and kidney, and increased urinary excretion in those Wistar rats. The dietary sesamin also elevated the hepatic mRNA levels of cytochrome P450 (CYP) 2B.
هيىلح صم لود ناطع لغطلا ميمص يف هيعجرم يفىطج ثيراي سيىك عجارملا يعجارجو
ةمذخحسملا تاماخلاو قرطلا يف عجارم ثحبلل ثفض ىولا ةضاطهلا ثحب ثعاحب تاماخلاو قرطلا هعضاو ىحىك كولا
ميقرحلا همزل شىلم هيذجبلاا فورحلاب هييضام اىحا عجارملا ميقرج شلابو هقذلا ىجرا
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