EFFECT OF OLIVE AND STEVIA LEAVES AND FLAXSEED EXTRACTS ON DIABETIC RATS

Document Type : Original Article

Abstract

ABSTRACT
Searching for effective, low cost and less side effect containing
herbal hypoglycemic agents is important therefore, the present study was
performed to evaluate the effect of olive and stevia leaves, flaxseed of
alcoholic extracts on diabetic rats. A total of forty eight male healthy rats,
weighing (201±1.5g) were classified in to 3 main groups of extraction of
(olive leaves, stevia leaves and flaxseed) each group was classified into 2
subgroup (levels of each type of extract at 0.2 and 0.5 %); in addition to
the control negative (-ve) and the control positive (+ve) groups. The rats
were divided into eight groups: Group (1), the control negative (-ve) was
fed on the basal diet. Group (2), the control positive (+ve) (diabetic rats)
was fed on the basal diet. Groups (3 and 4) are diabetic rats fed on the
basal diet containing olive leaves extract at 0.2 and 0.5 % respectively.
Groups (5 and 6) are diabetic rats groups fed on the basal diet containing
stevia leaves extract at 0.2 and 0.5 % respectively. Groups (7 and 8), are
diabetic rats groups were fed on the basal diet containing flaxseed extract
0.2 and 0.5 % respectively.
At the end of the experimental period (6 weeks), rats were
sacrificed and blood samples were collected to obtain serum. The results
indicated that, STZ treated rats showed significant increase in glucose
levels compared to normal rats. Supplementation with different levels of
olive leaves, stevia leaves and flaxseed in the diet caused significant
decrease (P<0.05) in the concentration of glucose level compared to the
positive control one. It was also observed that the concentration of serum
lipid profile, liver functions and kidney functions were significantly
improved for all tested groups, compared to the positive control group. In
conclusions, diet supplemented olive leaves, stevia leaves and flaxseed
caused an improvement of the biochemical results from diabetes,
therefore olive leaves, stevia leaves and flaxseed could be used as a
suitable supplementation therapy for diabetic patients.

Highlights

تأثیر المستخمص الکحولی لأو ا رق الزیتون ، الاستیفیا وبذور الکتان عمی الفئ ا رن
المصابة بالسکر
محمد صبری محمود* ، عید ذکی** ، میادة سعید محمد الحجار *
*: قسم التغذیة وعموم الاطعمة – کمیة الاقتصاد المنزلی- جامعة حموان
**: کمیة التربیة النوعیة – جامعة بنها
من المهم البحث عن نباتات فعالة ومنخفضة التکمفة واقل فی إحداث آثار جانبیة فی
تقمیل سکر الدم ، لذلک أجریت الد ا رسة الحالیة لتقییم تأثیر المستخمصات الکحولیة لأو ا رق
الزیتون والإستیفیا وبذور الکتان عمى الفئ ا رن المصابة بمرض السکری. تم تقسیم ثمانیة وأربعین
2.5 جم( إلى 3 مجموعات رئیسیة من ± من ذکور الفئ ا رن السمیمة البالغة ، وزنها ) 102
)أو ا رق الزیتون و الاستیفیا وبذور الکتان( تم تقسیم کل مجموعة إلى مجموعتین فرعیتین
%0.5 لکل مستخمص( ؛ بالإضافة إلى مجموعة الضابطة السالبة والمجموعة ،%0.1(
الضابطة الموجبة . قسمت الفئ ا رن إلى ثمانی مجموعات: المجموعة ) 2( المجموعة الضابطة
السالبة تم تغذیتها عمى النظام الغذائی الأساسی. المجموعة ) 1( المجموعة الضابطة الموجبة
، )الفئ ا رن المصابة بمرض السکری( تم تغذیتها عمى النظام الغذائی الأساسی. المجموعتان ) 3
4( عبارة عن فئ ا رن مصابة بداء السکری تتغذى عمى النظام الغذائی الأساسی الذی یحتوی عمى
6( عبارة عن ، مستخمص أو ا رق الزیتون بنسبة 0.1 و 0.5 ٪ عمى التوالی. المجموعتان ) 5
فئ ا رن مصابة بداء السکری تتغذى عمى النظام الغذائی الأساسی الذی یحتوی عمى مستخمص
8( من الفئ ا رن ، أو ا رق الاستیفیا بنسبة 0.1 و 0.5 ٪ عمى التوالی. تم تغذیة المجموعتین ) 7
Egypt. J. of Appl. Sci., 35 (7) 2020 67
20
المصابة بداء السکری عمى النظام الغذائی الاساسی المحتوی عمى مستخمص بذور الکتان 0.1
و 0.5 ٪ عمى التوالی .فی نهایة فترة التجربة ) 6 أسابیع( تم ذبح الفئ ا رن وتم جمع عینات الدم
لمحصول عمى سیرم الدم.
أظهرت زیادة معنویة فی مستویات STZ أشارت النتائج إلى أن الفئ ا رن المعالجة بمادة
الجموکوز مقارنة بالفئ ا رن السمیمة. التدعیم بمستویات مختمفة من أو ا رق الزیتون وأو ا رق الإستیفیا
فی ترکیز الجموکوز مقارنة بالمجموعة (P < وبذور الکتان أدی الی انخفاض معنوی ( 0.05
الضابطة الموجبة. کما لوحظ أن ترکیز الدهون فی الدم ووظائف الکبد ووظائف الکمى قد
تحسنت بشکل ممحوظ لجمیع المجموعات المختبرة ، مقارنة بالمجموعة الضابطة الموجبة.
الخلاصة: التدعیم باو ا رق الزیتون وأو ا رق الاستیفیا وبذور الکتان أدی الی تحسین النتائج
البیوکیمیائیة لمرض السکری ، لذلک یمکن استخدام أو ا رق الزیتون وأو ا رق الاستیفیا وبذور الکتان
کعلاج مکمل مناسب لمرضى السکری.

Keywords


EFFECT OF OLIVE AND STEVIA LEAVES AND
FLAXSEED EXTRACTS ON DIABETIC RATS
Mohamed S. Abd El-Baky* ; Eid A. Zaki **and Mayada S. El Haggar*
* Nutrition and Food Science Dept., Faculty of Home Economics, Helwan University
** Faculty of specific education, Banha University
Key Words: Diabetes, olive leaves, stevia leaves, flaxseed, glucose.
ABSTRACT
Searching for effective, low cost and less side effect containing
herbal hypoglycemic agents is important therefore, the present study was
performed to evaluate the effect of olive and stevia leaves, flaxseed of
alcoholic extracts on diabetic rats. A total of forty eight male healthy rats,
weighing (201±1.5g) were classified in to 3 main groups of extraction of
(olive leaves, stevia leaves and flaxseed) each group was classified into 2
subgroup (levels of each type of extract at 0.2 and 0.5 %); in addition to
the control negative (-ve) and the control positive (+ve) groups. The rats
were divided into eight groups: Group (1), the control negative (-ve) was
fed on the basal diet. Group (2), the control positive (+ve) (diabetic rats)
was fed on the basal diet. Groups (3 and 4) are diabetic rats fed on the
basal diet containing olive leaves extract at 0.2 and 0.5 % respectively.
Groups (5 and 6) are diabetic rats groups fed on the basal diet containing
stevia leaves extract at 0.2 and 0.5 % respectively. Groups (7 and 8), are
diabetic rats groups were fed on the basal diet containing flaxseed extract
0.2 and 0.5 % respectively.
At the end of the experimental period (6 weeks), rats were
sacrificed and blood samples were collected to obtain serum. The results
indicated that, STZ treated rats showed significant increase in glucose
levels compared to normal rats. Supplementation with different levels of
olive leaves, stevia leaves and flaxseed in the diet caused significant
decrease (P<0.05) in the concentration of glucose level compared to the
positive control one. It was also observed that the concentration of serum
lipid profile, liver functions and kidney functions were significantly
improved for all tested groups, compared to the positive control group. In
conclusions, diet supplemented olive leaves, stevia leaves and flaxseed
caused an improvement of the biochemical results from diabetes,
therefore olive leaves, stevia leaves and flaxseed could be used as a
suitable supplementation therapy for diabetic patients.
Egypt. J. of Appl. Sci., 35 (7) 2020 49-68
INTRODUCTION
Diabetes mellitus (DM) is one of the world’s fastest growing
chronic diseases. As the prevalence of diabetes continues to rise, it will
eventually reach a global rise by 2025 (Khavandi, et al., 2013).
According to the International Diabetes Foundation (IDF), there are 415
million people with diabetes in the world. By 2040, the number of
diabetes will increase to 642 million worldwide. The greatest increase in
the incidence and prevalence of diabetes is in the African population,
which is attributed to the dietary habits associated with urbanization and
westernization (Mennen, et al., 2001). Diabetes increases the risk of
developing heart diseases (Danesh et al., 2007).
Most of the oral drugs are costly and have a lot of side-effects.
Moreover, several medicinal plants, used to control diabetes along with
life style management, have been investigated for their beneficial effect
in different types of diabetes and are being more desired, owing to lesser
side-effects and low cost. In addition, during the past few years, many
phytoconstituents responsible for antidiabetic effects have been isolated
from hypoglycaemic plants. There are about 200 pure compounds from
plant sources reported to show blood glucose lowering activity. The
compounds may be flavonoids, alkaloids, carbohydrates, glycosides,
steroids, peptides and amino acids, phenolics and glycopeptides (Lo and
Wasser, 2011). Consumers across the world are becoming more
interested in foods with health promoting features as they gain more
awareness of the links between food and health. Over the last decades,
consumer demands for functional foods as an opportunity to improve
food product quality has increased enormously. The main
characterization of functional food is fortification with dietary fibre,
micronutrients, antioxidants, vitamins or minerals that contributes health
benefit effects in certain disorders. One of the promising functional
constituents that could be used for developing a functional food is inulin.
It has a great potential to be considered as a low glycaemic index (GI)
ingredient that could provide a number of health benefits such as
managing increased risk of chronic diseases (DM, cardiovascular
diseases, obesity, stroke and cancer), improving digestive health
(prevents constipation), reducing cholesterol and lipids and enhancing
mineral absorption from colon with its prebiotic role (prevents
osteoporosis) (Barclay et al., 2008).
The olive tree is mentioned in Quran and sunnah as a blessed tree,
and it is recommended to eat its fruit and use its oil, Many herbal
50 Egypt. J. of Appl. Sci., 35 (7) 2020
medicines have been recommended for the treatment of diabetes
(Chanwitheesuk et al., 2005). Hypoglycemic effects have been reported
for some plants that contain terpenoids, iridoid glycosides, flavonoids,
and other phenolic compounds (Li et al., 2004). In addition, a number of
secondary metabolites like flavonoids, phenolic acids, phenylpropanoids,
and terpenoids have shown significant antioxidant properties (Rahimi et
al., 2005; Harput et al., 2006 and Topcu et al., 2007). Historically,
olive leaves have been used as a folk remedy for combating diseases,
such as malaria (Garcia et al., 2000). In fact, Olive tree leaves can be
regarded as a particularly rich source of polyphenolic compounds, as
their polyphenolic contents may reach up to 40 g per kg of dry tissue.
A great number of medicinal plants have been used in the treatment
of diabetes in different part of the world. Evaluation of the antidiabetic
potentials of these plants becomes necessary to provide scientific proof
and justify their uses in ethnomedicine. Stevia rebaudiana belongs to the
Aster family which is indigenous to the northern region of South
America. It grows in many parts of Brazil, Paraguay, central America,
Thiland and Chine (Kolb et al., 2001).
Flaxseed oil is of particular interest in the nutritional therapy for
diabetes, given their potential role in several pathophysiological
processes related to cardiovascular disease (Jeppesen et al., 2013).
Recent studies suggest that for the vegetarians other than the fish oil
adding flaxseed oil to the diet may decrease insulin resistance in diabetics
and pre-diabetics and help in reducing the risk of developing type 2 as
well as type 1 diabetes (Bhardwaj et al., 2015).
Aim of the Study:
Searching for effective, low cost and less side effect containing
herbal hypoglycemic agents is important therefore, the present study was
performed to evaluate the effect of olive and stevia leaves, flaxseed of
alcoholic extracts on diabetic rats.
MATERIALS AND METHODS:
This study and all stages of experiment and analysis were
conducted at Graduate labs, Nutrition and Food Science Department,
Faculty of Home Economics, Helwan University. It was also be hiring
some specialized laboratories to conduct some determinations and
examinations.
Materials:
Chemicals: Streptozotocin was obtained from Sigma Company. Kits for
biochemical analysis were purchased from Gama Trade Company for
Egypt. J. of Appl. Sci., 35 (7) 2020 51
Pharmaceutical and chemicals, Dokki, Egypt. Casein, vitamins, minerals,
cellulose and choline were obtained from El-Gomhoria Company, Cairo,
Egypt.
Animals: Forty eight Male Sprague-Dawley rats weighing (201±1.5g)
were purchased from Farm of experimental animals in Helwan, Egypt.
Preparation of the basal diet:
The basal was prepared of protein 14 % (casein ≥85%), corn oil
4%, vitamin mixture 1%, salt mixture 3.5%, cellulose 5%, sucrose 1%
and reminder was corn starch as mentioned (Reeves et al., 1993).
Preparation of extraction of (olive and stevia leaves & flaxseed) :
A mixture of ethanol and water (20 ml, 70:30 (v/v)) were added
to the olive leaves and stevea leaves and flaxseed powdered (1 g). The
mixture was lifted to stand for at least one week at room temperature in
the dark. Subsequently, the solution was filtered using a 0.45 μm filter
paper. The extract was dried at 45 °C in rotary evaporator to produce a
semisolid mass and stored in airtight containers in refrigerator below
10°C.
Experimental animal design:
Injection with Streptozotocin: Streptozotocin (STZ) was
dissolved in a citrate buffer (pH 4.4) with a concentration of 15 mg/ml.
All animals were fasted overnight and were injected intraperitoneally (IP)
with a low dose STZ (60 mg/kg b.w.) for induction of diabetes, after 4
days, blood samples were obtained from medial canthus of eyes of each
rat to estimate glucose levels. Serum glucose was 100.34±1.22 and
300.01±2.15 mg/dl in healthy and injected rats respectively. Three days
later, the level of the blood glucose was assessed and the level ≥250
mg/dl was considered as diabetic (Ghauri et al .,2020).
A total of forty eight male healthy rats, weighing (201±1.5g) were
classified in to 3 main groups of extraction of (olive leaves, stevia leaves
and flaxseed) each group was classified into 2 subgroup (Levels of each
type of extract at 0.2 and 0.5 %); in addition to the control negative (-ve)
and the control positive (+ve) groups. The rats were divided into eight
groups: Group (1): The control (-ve) was fed on the basal diet. Group
(2): The control (+ve) (diabetic rats) was fed on the basal diet. Groups
(3 and 4): Diabetic rats groups were fed on the basal diet containing
olive leaves extract at the level of 0.2 and 0.5 % respectively. Groups (5
and 6): Diabetic rats groups were fed on the basal diet containing stevia
leaves extract at the level of 0.2 and 0.5 % respectively. Groups (7 and
52 Egypt. J. of Appl. Sci., 35 (7) 2020
8): Diabetic rats groups were fed on the basal diet containing flaxseed
extract at the level of 0.2 and 0.5 % respectively.
Blood sampling: At the end of experiment (4 weeks) rats were starved
for 12 hr., then sacrificed under ether anesthesia. Blood samples were
collected from the aortic vein into clean dry centrifuge tubes and stored
at room temperature for 15 minutes, put into a refrigerator for 2 hour,
then centrifuged for 15 minutes at 3000 rpm to separate serum. Serum
was carefully aspirated and transferred into dry clean Wasser –man tubes
by using a Pasteur pipette and kept frozen at (-20c) until analysis.
Biological Evaluation: Biological evaluations were carried out by
determination of feed intake (FI) which was recorded every day
throughout the experimental period. Body weight gain% (BWG) and feed
efficiency ratio (FER) were determined according to Chapman et al.,
1959, using the following equations:
BWG% =
× 100
FER = Weight gain (g) / Feed intake (g)
Biochemical analysis: For each group analyses included the following:
Serum total cholesterol (TC) were determined according to Allen
(1974).Triglycerides (TG) was done according to Fassati and Prencipe
(1982).High density lipoprotein–cholesterol (HDL-c) was determined
according to Lopez (1977) , whereas low density lipoprotein–cholesterol
(LDL-c) and very low density lipoprotein were determined according to
Friedewable et al.,( 1972).
LDL-c = TC - (HDL-c + VLDL-c)
Determination of aspartate aminotransferase (AST) and alanine
aminotransferase (ALT) were determined according to Reitman and
Frankel, (1957). Serum alkaline phosphates (ALP) was carried out
according to Belfield and Goldberg (1971).Urea was determined
according to Pattn and Crouch (1977), the determination of Creatinine
was according to Henry (1974), whereas glucose was determined
according to Trinder (1959).
Statistical analysis: The results were expressed as mean ± standard error
(SE). The statistical analysis was carried out by using SPSS, PC
statistical software (Verion 18.0 SPSS Inc., Chieago, USA) using the
Dunk 'test multiple range post-hoc test. All differences were considered
significant if P-values were (P˂ 0.05) (SPSS, 1993).
RESULTS:
Rats injected with STZ had significant (P≤0.05) higher glucose
level compared to the control negative group Table (1). Feeding diabetic
rats on diet supplemented with (stevia, olive or flaxseed) extract at the
Egypt. J. of Appl. Sci., 35 (7) 2020 53
level of 0.2 and 0.5% caused a significant decrease (P<0.05) in the
elevated serum glucose level, compared to the control positive group. It
was clear that, there were significant differences (P<0.05) in glucose
level among the treated groups with different two levels of (stevia, olive
or flaxseed) extract. While no significant difference in glucose level
among the treated groups with extract of olive at the level of 0.5% and
flaxseed at the level of 0.2 was detected. The percent of glucose
reduction as a result of supplementation with stevia, olive or flaxseed
extract are (43.77%, 51.88, 21.33%, 28.97%, 30.86% and 39.13%)
respectively, as compared to the value of glucose level in the positive
control group. Supplementation with extract of stevia caused the highest
reduction in glucose level.
Table (1) Effect of stevia, olive and flaxseed extracts on serum
glucose of diabetic rats
Parameters
Groups
Glucose (mg/dl) % of glucose
reduction
Control –ve 105.25±5.12 g -
Control +ve 317.50±3.30 a -
Stevia extract 0.2 % 178.50±1.93 e 43.77
Stevia extract 0.5% 152.75±4.23 f 51.88
Olive extract 0.2% 249.75±3.54 b 21.33
Olive extract 0.5% 225.50±3.42 c 28.97
Flaxseed extract 0.2% 219.50±3.96 c 30.86
flaxseed extract 0.5% 193.25±3.17 d 39.13
Data are expressed as mean ± SE.
Means with different superscript letters in the column are significantly differences
at (P ≤ 0.05).
The results in Table (2) revealed the effect of (stevia, olive or
flaxseed) extract at the level of 0.2 and 0.5% on liver function of diabetic
rats. The activities of serum ALT and AST significantly increased
(P<0.05) in the diabetic group, compared with the corresponding value of
normal control group .
Supplementation with (stevia, olive or flaxseed) extract at the
level of 0.2 and 0.5% significantly decreased (P<0.05) the elevated levels
of both serum ALT and AST compared to the negative control group. It
was clear that, no significant difference in AST and ALT levels among
the treated groups with different two levels of stevia extract. While there
were significant differences (P<0.05) in serum AST among the treated
groups with different two levels of flaxseed extract. Moreover, there was
no significant difference in serum AST and ALT among the treated
groups, but extract of olive at the level of 0.5% were significantly
different (P<0.05) among the treated groups. Supplementation with
54 Egypt. J. of Appl. Sci., 35 (7) 2020
extract of olive at the level of 0.5% caused the highest reduction in liver
function.
Table (2): Effect of stevia, olive and flaxseed extracts on liver
functions of diabetic rats
Parameters
Groups
AST (μ/L) ALT (μ/L)
Control –ve 83.00±2.88 d 40.20±2.16 d
Control +ve 119.60±2.58 a 99.67±0.95 a
Stevia extract 0.2 % 104.57±4.33 bc 74.25±2.80 b
Stevia extract 0.5% 103.37±1.74bc 72.35±2.83 b
Olive extract 0.2% 99.42±4.46 c 69.07±3.36 b
Olive extract 0.5% 87.80±3.20 d 59.00±3.53 c
Flaxseed extract 0.2% 109.65±1.43 b 75.75±3.06 b
flaxseed extract 0.5% 98.62±1.46 c 73.42±3.93 b
Data are expressed as mean ± SE.
Means with different superscript letters in the column are significantly differences
at (P ≤ 0.05).
Table (3) illustrates the effects of (stevia, olive or flaxseed)
extract at the level of 0.2 and 0.5% in serum kidney functions on diabetic
rats. Injection with STZ significantly increase (P<0.05) the level of
creatinine, urea and uric acid, compared to the control normal group
(control –ve). Feeding diabetic rats on diet supplemented with stevia,
olive or flaxseed extract at the tested level caused a significant decrease
(P<0.05) in the mean values of creatinine, urea and uric acid as compared
to the positive control group. There were no significant differences in
serum creatinine, urea and uric acid between the treated groups.
Table (3): Effect of stevia, olive and flaxseed extracts on kidney
functions of diabetic rats
Parameters
Groups
Creatinine Urea Uric acid
mg/dl
Control –ve 0.325±0.08 c 48.35±1.55 d 2.77±0.13 c
Control +ve 1.25±0.15 a 96.50±3.01 a 6.27±0.34 a
Stevia extract 0.2 % 0.825±0.08 b 78.12±2.13 b 4.15±0.40 b
Stevia extract 0.5% 0.750±0.06 b 72.85±4.50 b 4.17±0.41 b
Olive extract 0.2% 0.750±0.06 b 70.12±4.00 bc 3.97±0.16 b
Olive extract 0.5% 0.625±0.07 b 59.77±2.39 c 3.77±0.14 b
Flaxseed extract 0.2% 0.850±0.06 b 77.07±6.57 b 4.27±0.27 b
flaxseed extract 0.5% 0.800±0.09 b 69.23±2.21 bc 4.10±0.55 b
Data are expressed as mean ± SE.
Means with different superscript letters in the column are significantly differences
at (P ≤ 0.05).
Results illustrated in Table (4) shows the effect of (stevia, olive or
flaxseed) extract on lipids profile of diabetic rats. STZ injection to rats
caused a significant increase (P<0.05) in serum lipid profile, however, serum
Egypt. J. of Appl. Sci., 35 (7) 2020 55
HDL-C was significantly lowered, compared to the healthy rats. Diet
supplemented with stevia, olive or flaxseed extract at the level of 0.2 and
0.5% significantly decrease (P<0.05) the mean value of serum TC, TG,
VLDL-C and LDL-C, however, serum HDL-C level was increased
significantly (P<0.05), compared to the positive control group.
It was clear that, no significant difference in TC, TG and VLDL
levels but there were significant differences (P<0.05) in HDL and LDL
levels among the treated groups with different two levels of stevia extract.
Moreover, no significant difference in lipid profile among the treated groups
with different two levels of olive extract. Also, no significant difference in
TG and VLDL levels but there were significant differences (P<0.05) in TC,
HDL and LDL levels among the treated groups with different two levels of
flaxseed extract. There were no significant differences in serum TC between
different two levels of (stevia and olive) extract. It was obvious that, the
treatments with flaxseed extract at the level of 0.5% gave the highest
beneficial effect in improving lipid profile in diabetic rats.
Regarding to changes in body weight status, Table (5) illustrated the
changes of body weight, feed intake and FER in the diabetic rats fed on diet
supplemented with of (stevia, olive or flaxseed) extract. The initial body
weight of rats was (201.50±1.85 g), there were no significant differences in
IBW among all groups. Diabetic rats had significant decrease (P<0.05) in
the FBW compared to the negative control group. It was observed that STZ
induced diabetic in rats caused significant decrease (P<0.05) in FBW
compared to the healthy rats .
The supplementation with (stevia, olive or flaxseed) extract
significantly (P<0.05) increased the lowered FBW compared to the positive
control group. There were significant differences (P<0.05) in FBW among
the treated groups . There were significant differences (P<0.05) in FBW
among the treated groups with different two levels of Stevia extract, while
no significant change in FBW among the treated groups with different two
levels of (olive or flaxseed) extract.
Stevia extract at the level of 0.5% caused the highest increase in FBW
compared to other treatments. In regarding to BWG% and FER, diabetic rats
had significantly (P<0.05) lowered BWG% and FER but a significant increase
in FI compared to the negative control group. There were no significant
differences in BWG% and FER between the groups fed on (0.2 and 0.5%) of
(olive or flaxseed) extract, while the supplementation with Stevia extract
showed significant differences (P<0.05) in BWG%. However, the
supplementation with the tested materials caused a significant increase (P<0.05)
in BWG% and FER compared to the positive control group, while no
significant change in FER among the treated groups with different two levels of
stevia extract. Stevia extract at the level of 0.5% caused the highest increase in
BWG% and FER compared to other treatments.
56 Egypt. J. of Appl. Sci., 35 (7) 2020
Table (4): Effect of stevia, olive and flaxseed extracts on lipid profile of diabetic rats
Parameters TC TG HDL-C VLDL-C LDL-C
Groups (mg/dl)
20.55±2.61e 16.17±0.99 f 59.25±2.39 a 80.87±4.95 f 95.97±3.56 e Control –ve
88.45±3.81 a 26.27±0.35 a 25.75±1.10 d 131.35±1.78 a 140.47±2.57 a Control +ve
68.68±11.88 b 24.36±0.55 b 34.75±1.70 c 121.82±2.76 b 127.80±1.87 b Stevia extract 0.2 %
53.91±2.23 c 23.76±0.27 bc 45.25±1.25 b 118.82±1.35 bc 122.92±3.19 bc Stevia extract 0.5%
51.12±3.28 c 22.30±0.57 cd 47.92±2.71 b 111.50±2.87 cd 121.35±1.42 bc Olive extract 0.2%
51.25±1.20 c 20.90±0.45 de 48.75±1.75 b 104.50±2.25 de 120.90±1.82 bc Olive extract 0.5%
46.18±2.76 c 20.16±0.83 e 49.75±1.31 b 100.82±4.16 e 116.10±1.71 c Flaxseed extract 0.2%
33.46±2.64 d 19.66±0.73 e 55.96±1.63 a 98.32±3.67 e 109.10±1.00 d flaxseed extract 0.5%
Data are expressed as mean ± SE.
Means with different superscript letters in the column are significantly differences at (P ≤ 0.05)
Table (5) Effect of stevia, olive and flaxseed extracts on FI, BWG and FER of diabetic rats
Parameters IBW (g) FBW (g) BWG% FI (g/d/rat) FER
Groups
0.030±0.02 a 19.93±1.60 22.00 a 239.86±2.42 a 200.11±1.91 a Control –ve
- 0.073±0.04 e - 29.06±1.77 13.30 f 142.80±3.00 e 201.50±1.85 a Control +ve
0.021±0.01 bc 12.99±1.10 20.00 c 226.67±2.41 b 200.81±3.50 a Stevia extract 0.2 %
0.026±0.007 ab 16.39±0.48 21.00 b 237.53±1.99 a 204.10±1.81 a Stevia extract 0.5%
0.015±0.01 cd 8.47±0.67 18.40 de 216.60±1.86 cd 199.73±2.21 a Olive extract 0.2%
0.018±0.006 cd 10.30±0.41 19.00 cd 222.61±1.51 bc 201.85±1.78 a Olive extract 0.5%
0.012±0.01d 6.53±0.62 17.00 e 211.83±0.67 d 198.90±1.42 a Flaxseed extract 0.2%
0.014±0.01 d 7.68±0.75 17.50 de 216.77±2.26 cd 201.30±1.51 a flaxseed extract 0.5%
Data are expressed as mean ± SE.
Means with different superscript letters in the column are significantly differences at (P ≤ 0.05)
Egypt. J. of Appl. Sci., 35 (7) 2020 57
10
DISCUSSION:
DM is a chronic condition that grows the most, especially in
developing countries. The disease is highlighted for the severity of its
complications (Whiting et al., 2011). Controlling the blood glucose
level is vital for protecting complications and improving the diabetics’
health (Ceriello, 2005). The available hypoglycemic drugs have
numerous limitations, therefore herbal medicine can help to minimize
diabetic complications.
In the present study feeding diabetic rats on diet supplemented
with olive, stevia leaves and flaxseed extracts caused a significant
decrease (P<0.05) in the elevated serum glucose level, compared to the
control positive group. These results are in agreement with a study
carried out by Azzawie and Alhamdani, (2006) who found that ethanol
extract of olive leaf decreased blood glucose with diabetic rabbits. In the
study by pudyal et al., (2010) showed that treatment with OLE lowered
blood glucose levels and improved glucose tolerance in OLE rats
compared with control group. Another study examined the effect of 500
mg olive leaf extract on both diabetic patients and streptozotocin-induced
diabetic rats for 1 week , the results indicated a significant decreased in
blood glucose (Wainstein et al., 2012).
The anti-hyperglycemic effect of S. rebaudiana was investigated
in both rats and humans by (Jeppesen et al., 2003 and Thomas, 2010).
Ahmad and Ahmad, (2018) showed that stevia extract decrease blood
glucose level in treated diabetic rats, compared with the diabetic and nondiabetic
control rats after 8 weeks. The aqueous extract of stevia has antidiabetic
effects in albino rats, and therefore could be promising
nutraceutical therapy for the management of diabetes and its associated
complications.
Flaxseed may delay the development of T2DM in Zucker rats,
hypoglycemic effect due to its antioxidant activity (Prasad, 2000).
Flaxseed consumption can reduce the speed of glucose absorption and
the need for insulin production (Morisset et al., 2009). Soltanian and
Janghorbani, (2018) mentioned that flaxseed supplementation reduced
blood glucose in subjects with type 2 diabetes. Hasaniani et al., (2019)
showed that the addition of flaxseed to yogurt can be effective in the
management of type 2 diabetes. Moreover, Abduljawad, (2019)
mentioned that supplementation with flaxseed powder in the diet was
more effective than flaxseed oil (4%) in the reduction of blood glucose
level of diabetic rats. Yari et al., (2016) have shown that flaxseed
58 Egypt. J. of Appl. Sci., 35 (7) 2020
11
supplementation in patients with metabolic syndrome decreased insulin
resistance and FBS level.
Jemai et al., (2009) reported that 8 and 16 mg/kg doses of olive
leaf extract decreased serum glucose level. Olive leaf extract was able to
cure glucose metabolism in liver and kidneys of rats by minimizing
oxidative stress in rats (Salah et al., 2013). Abunab et al., (2017) found
that olive leaf extract decreased blood glucose level in diabetic rats.
Similarly, Ajwad et al., (2020) showed that the diabetic rats treated with
olive extract showed reduction in fasting blood glucose by enhanced
insulin sensitivity, improved some antioxidative parameters. These
results imply that olive leaf extract may have a potential effect in the
treatment of Type 2 diabetes and prevention of its complications (Acar-
Tek and Ağagündüz 2020). Khattab, et al (2020) showed that olive
leave extract may exert a protective role against STZ induced diabetic
nephropathy via an antioxidant mechanism.
Regarding to liver functions, Al-Sahib and Alsaadi, (2020)
showed that 4% and 8% olive leaves improved ALT and AST enzymes
activities compared to (diabetic) positive control group. The decrease in
the liver enzymes may be due to the presence of some active constituent
like flavonoids and terpenoids in the OL which have hepatoprotective
effect against hepatotoxins (Miles et al., 2005). Fki et al., (2020) suggest
that the oleuropein- and hydroxytyrosol-rich olive leaf extracts possessed
hypolipidemic and hepatoprotective effects against the HFD-induced
metabolic disorders. These results are in the same line with the obtained
results.
Shivanna et al., (2013) showed a reduction of blood glucose,
ALT and AST in the stevia whole leaves powder. Assaei et al., (2016)
suggested that supplementation with an aquatic extract of stevia (400
mg/kg) for the period of 28 days significantly reduced triglycerides,
ALT, AST levels, Creatinine, Urea and Uric acid compared with
diabetic rats (p<0.05). So stevia acts exerts beneficial antihyperglycemic
effects through stevia’s antioxidant properties.
Hyperproteinemia and hyperalbuminemia associated with
dysfunctions of liver and kidney, and the increased rate of body water
loss (Kalaiselvi et al., 2015).These observations are generally in
agreement with other investigations on STZ and alloxan induce relative
influences (Abdel-Kader et al., 2019 and Al-Attar and Alsalmi, 2019).
Egypt. J. of Appl. Sci., 35 (7) 2020 59
12
Dietary flaxseed has protective effects on the kidney in some
animal models of chronic renal disease (Ogborn et al., 1999) and in
humans with lupus nephritis (Clark et al., 2001).
Laaboudi et al., (2016) suggested that olive tree extract
consumption can improve lipid profile, and reduce glycemia. It was
found that olive oil improve lipid profiles and blood glucose in type-2
diabetic patients through the effect of monounsaturated fatty acids
(MUFA), the minor components of olive oil which prevents central fat
redistribution and the postprandial decrease in peripheral adiponectin
gene expression and insulin resistance induced by a carbohydrate-rich
diet in insulin-resistant subjects (Al Jamal and Ibrahim, 2011).
Diabetes is associated with fundamental changes in serum lipids profile
(Ani and Aginam, 2018), that is mainly attributable to increased TC and
TG levels and impaired lipoprotein profile (Ormazabal et al., 2018).
Long-term oral consumption of olive leaves by streptozocininduced
diabetic rats has been hypoglycemic effect, decreases serum
triglyceride, total cholesterol and LDL cholesterol and increases HDL
level (Basuny and Mohammed, 2020).
Haliga et al., (2015) suggested that the high dose of ground
flaxseed reduced total cholesterol and increasing HDL‑C levels. Bassett
et al., (2017) suggested that flaxseed can also lower circulating trans fats
levels. Flaxseed may lower circulating cholesterol levels (Parikh et al.,
2019). Abduljawad, (2019) mentioned that 10% flaxseed powder
decreases serum triglyceride, total cholesterol and LDL cholesterol and
increases HDL level as compared to diabetic control group.
Al-Attar and Alsalmi, (2019) noticed the extract of olive leaves
(200 and 400 mg/kg body weight) was investigated for antidiabetic
activity on STZ diabetic rats. Significant declines of body weight gain in
STZ-diabetic rats were noted after eight weeks. Similar observations
were noted in many experimental diabetes researches (Zhang et al., 2016
and Abdel-Kader et al., 2019). Also, Jung et al., (2019) suggest that
OLE is useful for preventing or treating obesity in which the
administration of the oleuropein- and hydroxytyrosol-rich olive leaf
extracts identically reduced the body weight, the weight gain, and the
epididymal fat accumulation.
Ahmad and Ahmad, (2018) showed that stevia aqueous extract
(200, 300, 400 and 500 ppm/kg b.w) for 8 weeks improved weight
control in diabetic rats by decreasing the feed intake and body weight
gain.
60 Egypt. J. of Appl. Sci., 35 (7) 2020
13
Regimen diet containing supplemented bakery with flaxseed or
flaxseed oil for 12 weeks decrease BMI, blood glucose, lipid profile in
type 2 diabetics (Tharwat et al., 2017). Soltanian and Janghorbani,
(2018) suggested that flaxseed cookies (10 g of flaxseed pre-mixed in
cookies twice per day) used as a snack may be a useful tool for
decreasing, weight, glycemic and lipid levels.
Conclusion: Olive and stevia leaves and flaxseed have long been utilized
for their various beneficial effects in traditional medicine. There are still
a long way ahead of these plants and their compounds to find their place
among drugs actively used in modern medicine to alleviate blood
glucose, their remarkable health-benefitting effects should not be
overlooked, which are very much worthy of further investments and
investigations.
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تأثیر المستخمص الکحولی لأو ا رق الزیتون ، الاستیفیا وبذور الکتان عمی الفئ ا رن
المصابة بالسکر
محمد صبری محمود* ، عید ذکی** ، میادة سعید محمد الحجار *
*: قسم التغذیة وعموم الاطعمة – کمیة الاقتصاد المنزلی- جامعة حموان
**: کمیة التربیة النوعیة – جامعة بنها
من المهم البحث عن نباتات فعالة ومنخفضة التکمفة واقل فی إحداث آثار جانبیة فی
تقمیل سکر الدم ، لذلک أجریت الد ا رسة الحالیة لتقییم تأثیر المستخمصات الکحولیة لأو ا رق
الزیتون والإستیفیا وبذور الکتان عمى الفئ ا رن المصابة بمرض السکری. تم تقسیم ثمانیة وأربعین
2.5 جم( إلى 3 مجموعات رئیسیة من ± من ذکور الفئ ا رن السمیمة البالغة ، وزنها ) 102
)أو ا رق الزیتون و الاستیفیا وبذور الکتان( تم تقسیم کل مجموعة إلى مجموعتین فرعیتین
%0.5 لکل مستخمص( ؛ بالإضافة إلى مجموعة الضابطة السالبة والمجموعة ،%0.1(
الضابطة الموجبة . قسمت الفئ ا رن إلى ثمانی مجموعات: المجموعة ) 2( المجموعة الضابطة
السالبة تم تغذیتها عمى النظام الغذائی الأساسی. المجموعة ) 1( المجموعة الضابطة الموجبة
، )الفئ ا رن المصابة بمرض السکری( تم تغذیتها عمى النظام الغذائی الأساسی. المجموعتان ) 3
4( عبارة عن فئ ا رن مصابة بداء السکری تتغذى عمى النظام الغذائی الأساسی الذی یحتوی عمى
6( عبارة عن ، مستخمص أو ا رق الزیتون بنسبة 0.1 و 0.5 ٪ عمى التوالی. المجموعتان ) 5
فئ ا رن مصابة بداء السکری تتغذى عمى النظام الغذائی الأساسی الذی یحتوی عمى مستخمص
8( من الفئ ا رن ، أو ا رق الاستیفیا بنسبة 0.1 و 0.5 ٪ عمى التوالی. تم تغذیة المجموعتین ) 7
Egypt. J. of Appl. Sci., 35 (7) 2020 67
20
المصابة بداء السکری عمى النظام الغذائی الاساسی المحتوی عمى مستخمص بذور الکتان 0.1
و 0.5 ٪ عمى التوالی .فی نهایة فترة التجربة ) 6 أسابیع( تم ذبح الفئ ا رن وتم جمع عینات الدم
لمحصول عمى سیرم الدم.
أظهرت زیادة معنویة فی مستویات STZ أشارت النتائج إلى أن الفئ ا رن المعالجة بمادة
الجموکوز مقارنة بالفئ ا رن السمیمة. التدعیم بمستویات مختمفة من أو ا رق الزیتون وأو ا رق الإستیفیا
فی ترکیز الجموکوز مقارنة بالمجموعة (P < وبذور الکتان أدی الی انخفاض معنوی ( 0.05
الضابطة الموجبة. کما لوحظ أن ترکیز الدهون فی الدم ووظائف الکبد ووظائف الکمى قد
تحسنت بشکل ممحوظ لجمیع المجموعات المختبرة ، مقارنة بالمجموعة الضابطة الموجبة.
الخلاصة: التدعیم باو ا رق الزیتون وأو ا رق الاستیفیا وبذور الکتان أدی الی تحسین النتائج
البیوکیمیائیة لمرض السکری ، لذلک یمکن استخدام أو ا رق الزیتون وأو ا رق الاستیفیا وبذور الکتان
کعلاج مکمل مناسب لمرضى السکری.
68 Egypt. J. of Appl. Sci., 35 (7) 2020

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