1. health and fitness
2. disease
3. cholesterol

Pakistan Journal of Pharmacology
Vol.28, No.1, January 2011, pp.1-8
HYPOGLYCEMIC AND HYPOCHOLESTEROLEMIC EFFECTS
OF ACACIA TORTILIS (FABACEAE) GROWING IN MAKKAH
WAHEEB DAKHELALLAH MOHAMMAD ALHARBI
AND AISHA AZMAT
Department of Physiology, Faculty of Medicine
Umm-Al-Qura University Makkah, Saudi Arabia
ABSTRACT:
Traditional use of herbs and plants is centuries old and they have shown therapeutic
properties. But scientific studies are not done to confirm their efficacy and margin of
safety. Acacia was collected from UQU, Makkah. Leaves were separated from shoots and
aqueous extract of Acacia tortilis was prepared at the concentration of 100mg/ml. Acacia
aqueous extract was administered at the dose of 800mg/kg in treated rats for seven days.
The various parameters studied include blood glucose levels, serum cholesterol levels,
HDL, LDL, AST, ALT and Urea. All parameters were estimated using diagnostic kits
spectrophotometrically. At the end of study autopsy also performed. Administration of
Acacia aqueous extract (800mg/kg) for seven consecutive days caused significant (P<
0.05) decrease in blood glucose, serum total cholesterol, and LDL level while an increase
in serum HDL- cholesterol was observed as compared to their control rats. However a
statistically significant (P< 0.05) decrease in body weight was also observed. Autopsy
did not show any sign of abnormality and all rats were comparable with their control rats.
The findings of the present study confirm that above-mentioned plant validates our claim.
In the light of our pharmacological and toxicological studies, Acacia tortilis appears to be
a valuable plant, which can be useful, at least as an adjunct, in the therapy of diabetes and
hypercholesterolemia or in a condition in which hyperglycemia and dyslipidemia coexist
quite often. Further the toxicological, hematological and autopsy results further confirm
the wide marginal safety of Acacia tortilis leaves aqueous extract.
Keywords: Acacia tortilis leaves aqueous extract, Hypoglycemia, Hypocholesterolemia,
Flavanoids.
INTRODUCTION
Hypercholesterolemia
and
Diabetes
mellitus (DM) are the commonest disorder that
affects more than 100 million people
worldwide (6% of the population) (WHO/
Acadia, 1992). A previous population-based
national epidemiological study conducted
among adult Saudi (age 15 years) reported that
the risk of developing hyper-cholesterolemia
increased by 2% for each year (Al-Nuaim,
1997). Hypercholesterolemia, high levels of
serum triglycerides (TG) are strong,
independent predictors for CHD (Hokanson &
Austin, 1996 and Haim et al., 1999). Hypercholesterolemia is an important risk factor for
atherosclerosis, myocardial infarction or stroke
(Morozova et al., 2005), While DM is found to
damage many of the body systems,
particularly the blood vessels and nerves
(Nagappa et al., 2003). For its therapy, various
clinical practice guidelines have addressed the
treatment of hypercholesterolemia. The
American College of Physicians has addressed
hypercholesterolemia in patients with, diabetes
(Snow et al., 2004). However, along with the
synthetic drugs, a change in diet (Tang et al.,
1998), weight management, physical activity
and many agents of the plant origin are also in
use. The National Center for Complementary
and Alternative Medicine focused on use of
complementary and alternative medicine
Correspondence to: e-mail: [email protected]
2
Hypoglycemic and hypocholesterolemic effects of Acacia tortilis
(CAM) suggest that CAM was used to treat
hypercholesterolemia by 1.1% of U.S. adults
during 2002 (Durrington, 2003). Plants are
always an exemplary source of drugs; in fact
many of the currently available drugs were
derived either directly or indirectly from them.
According to world ethnobotanical information reports, almost 800 plants may possess
antidiabetic and hypercholesterolemic activity
(Alarcon-Aguilara et al., 1998: Iman et al.,
2003; Metwally et al., 2009). Now in recent
years there has been a growing trend to
evaluate
the
plant
constituents
on
pharmacological basis and screening of more
effective and safe hypoglycemic and
hypocholesterolemic agents has continued to
be an important area.
Therefore the present work have been
planned to study indigenous plants (Acacia
tortilis) of Makkah using new and sensitive
technologies and statistical analysis. Acacia is
a genus of shrubs and trees belonging to the
subfamily Mimosoideae of the family
Fabaceae, first described from Africa by
Linnaeus in 1773. Acacia is one of the plant
that has been frequently used as medicine like
Acacia Bark has been used in treatment of
haemoptysis, gonorrhea (Wallis, 1967),
leprosy and headaches (Stacey and Michael
Chillemi, 2007). Acacia Catechu has been
used for treating fever, diarrhoea, leucorrhoea,
piles, erysipelas (Kirtikar, 1935), throat
infection (Usher, 1984). It is reported that
Acacia catechu has hypoglycaemic activity
(Singh et al., 1976) hepatoprotective,
antipyretic and digestive properties (Chopra et
al., 1956; Jayasekhar et al., 1997).
Considering the above facts, the present study
was undertaken to evaluate the hypocholesterolemic and hypoglycaemic activities
of Acacia in experimental animal models.
MATERIALS AND METHOD
Plant Material:
Acacia was collected from Umm-Al-Qura
University (UQU) in Alawali Makkah, Saudi
Arabia in May 2010 and the leaves were
separated from shoots and wood. The plant
was authenticated by plant taxonomist Dr.
Kadry Abdel Khalik, Associate Professor of
Plant Taxonomy, UQU, Faculty of Science,
Biology Department, Makkah, Saudi Arabia.
Extraction:
Fifty grams of leaves of Acacia were
washed with 1000 ml of distilled water and
rinsed twice with 500 ml of distilled water. To
prepare the water extract, 50g of leaves were
milled using the blender in 500 ml of distilled
water to obtain 100mg/ml. The extract mixture
was allowed to mix on shaking water bath at
40°C for half an hour and filtered. The
resultant solution (500 ml) or filtrate was then
refrigerated. This Acacia tortilis leaves
aqueous extract (AAq) used on each day of the
experiment. The extract solution was
administered orally post esophagus in a dose
of 800mg/1000 g of animal concentration of
extract 100mg/ml (Meena et al., 2006).
Animals:
Twenty Adult Sprague dawley rats (200250g) of either gender were obtained from
animal house of UQU and were housed in
groups of 3 per cage for seven days prior to
experimentation with free access to food and
tap water ad libitum and kept on a 12h
light/dark cycle. Each experimental group
consisted of ten animals. All animals were
housed in an air-conditioned room at 23+1 ºC
during the quarantine period.
Toxicological studies in rats:
Two groups of Sprague dawley rats (200250 g) containing ten animals in each group
were used in this study. All animals were
treated orally p.o. once daily for seven
consecutive days.
Group I (5 males and 5 females), was
treated with Acacia tortilis leaves aqueous
extract (AAq) (800 mg/kg), while Group II (5
males and 5 females) served as control
received distilled water only.
Waheeb Alharbi and Aisha Azmat
Estimation of different biochemical
Parameters and Autopsy:
At the end of 7th day all survived rats were
anaesthetized with ether and autopsied
(Mohamed et al., 2002). The blood samples
approximately (4 to 8ml) were withdrawn
from cardiac puncture before dissecting the
animals with sterile disposable syringe (HI
Med, Korea) from all treated and control rats,
were left at room temperature for 20 minutes.
Then incubated at 37oC for 30 minutes and
centrifuged separately at the speed of 3000
rpm for 20 minutes. Supernatants (Serum)
were separated out and the residue was
discarded. Serum obtained (1-3 ml) was
subjected for the study of following
parameters.
Cholesterol, Glucose, Lactate dehydrogenase
(LDH), High density lipoprotein (HDL),
ALT/GPT, AST/GOT and Urea.
All these kits were purchased from
Spinreact, S.A. Ctra. Santa Coloma, 7 E-17176
SANT ESTEVE DE BAS (GI) SPAIN.
Spectrophotometer (APEL, PD 303UV) and
Water bath (Gesellschaft fur Labortechnik
m.b.H and co D3006 Burgwedel 1) were used
to measure the absorbance of light and for
incubation.
3
Statistical analysis:
Changes in serum biochemical levels were
compared using student’s t-test. P<0.05 was
used as level of significance.
RESULTS
Hypoglycemic activity of Acacia (800mg/kg)
in rats:
The observations of present study showed
that the oral administration of AAq at the dose
of 800mg/kg showed a significant (p<0.05)
hypoglycemic effect (Table 1) in normoglycaemic rats that’s was 39% in comparison
with their control as shown in Fig 1.
Hypocholesterolemic activity of Acacia
(800mg/kg) in rats:
Oral administration of AAq (800mg/kg)
showed a fall in the serum cholesterol level
(17%) in treated rats, when compared to their
control values as shown in Fig 1. The
reduction in serum cholesterol level was
statistically significant (p<0.05) in comparison
with their respective controls. It prevented the
elevation of lipid profile significantly (Table
1).
After the administration of AAq (800
Fig. 1. Effect of AAq (800mg/kg) on glucose and lipid profile.
Data are expressed as Percent mean (%) ± SEM. Significant at *P < 0.05
4
Hypoglycemic and hypocholesterolemic effects of Acacia tortilis
mg/kg) a decrease was observed in serum LDL
level (31%) as shown in Fig 1. However, an
increase was observed in serum HDL level
that was 75% when compared to its control as
shown in Fig 1.
in body weight after 7th days treatment was
statistically significant (P<0.05). The body
weights of the rats before (on ‘0’ day) and
after (on 7th day) drug administration were
compared using Student’s‘t’ test (Paired).
Effect of Acacia on body weight:
Fig. 2 showed the effect of AAq
(800mg/kg) on body weight of rats. During the
study the changes in body weight were
recorded and compared with their respective
control. The results showed that AAq was
found to decrease body weight in both male
and female rats by 10% (Fig. 2). This decrease
Toxicological studies in rats:
Toxicological studies of Acacia were
carried out in rats. Oral administration of
Acacia (800 mg/kg) did not show any change
in physical behavior of rats. None of the
groups examined showed any mortality. None
of these animals showed any sign of toxicity
and change physical behavior after first 2 hour.
Fig. 2. Effect of AAq on body weight
Values are expressed as mean+ SE (n).
* Show P<0.05 indicate significantly different from control
Table-1
Effects of Acacia tortilis leaves (AAq) extract at the dose 800 mg /kg
on different biochemical parameter
Parameter
Cholesterol
Lactate dehydrogenase (LDH)
High density lipoprotein (HDL)
Glucose
Alanine transaminase (ALT)
Aspartate aminotransferase (AST)
Urea
Control
65.63+4.50 (10)
37.15+6.00 (10)
83.90+10.9 (10)
61.08+4.46 (10)
2.290+0.22 (10)
2.870+0.70 (10)
34.90+11.0 (10)
All the value have been presented as mean ± Standard error (n)
* Indicate level of significance P<0.05 when compared to control.
Treated
55.49+4.62 (10)*
23.23+2.18 (10)*
113.8+6.80 (10)*
39.67+7.23 (10)*
1.850+0.05 (10)
2.877+0.56 (10)
32.90+7.86 (10)
Waheeb Alharbi and Aisha Azmat
Effect of Acacia on different biochemical
parameters:
Oral administration of 800 mg/kg dose of
Acacia for 7 consecutive days was not found
to alter the liver, heart and kidney enzyme
significantly. It produced slight but nonsignificant decrease in the serum ALT/GPT as
shown in Table 1, but this decrease was also
statistically non significant (p>0.05) as shown
in Table 1. Heart profile showed that It
increase the serum AST/GOT in comparison to
control rats, although this increase was
statistically non significant (p>0.05), as shown
in Table 1. A non-significant decrease in the
serum urea was also observed in the AAq (800
mg/kg) treated rats in comparison to control
rats. These reductions were statistically non
significant (p>0.05) showing the normal
profile of kidney.
Autopsy:
Autopsy revealed that no gross changes
were observed in organs like liver, spleen,
heart and kidney. Drug did not cause any
internal body hemorrhage
DISCUSSION
In the present study rats were feed AAq
(800mg/kg) for 7 consecutive days. The
observations
showed
that
the
oral
administration of AAq tortilis at the dose of
800mg/kg showed significant (p<0.05)
hypoglycemic effect (Fig 1). The results
indicate the presence of some powerful
hypoglycemic activity which suppressed the
elevation of sugar level. Previously Singh et
al., 1976 showed that Acacia catechu extract
decreased blood sugar of normal rats. It is
believed that the extract causes the pancreas to
secrete more insulin in normal animals (Singh
et al., 1976). Another study showed the effect
of gum Arabica on carbohydrate metabolism
and they found it be beneficial to the diabetic
patient; it suppressed the diabetes during
feeding period only (Singhal et al., 1982).
Acacia coriacea also significantly reduced the
initial rise in plasma glucose levels (p<0.05)
and the area under the plasma glucose curve
(p<0.005) in six non-diabetic subjects
5
(Thorburn et al., 1987). Some other studies
also showed that flavonoids, alkaloids and
saponins which are present in Acacia tortilis
leaves have been documented to have blood
glucose reduction effect (Bolkent et al., 2000;
Diatewa et al., 2004). These findings were in
accordance with the present results confirming
the hypoglycemic effect of Acacia tortilis (Fig
1). In the present study AAq (800mg/kg)
induces hypocholesterolemia in normal rats. It
also prevented the elevation of lipid profile
significantly because administration of AAq
decrease the level of LDL and increase the
level of HDL significantly (Table 1).
The mechanism(s) of hypolipidemic effect
of the Acacia tortilis leaves may be similar to
some of those proposed previously by Stacey
and Michael Chillemi (2007), they suggest that
fruit is rich in essential fatty acids and is said
to be very beneficial for good coronary health.
β-sitosterol (beta sitosterol) and phytosterol
that competes with dietary cholesterol for
absorption, it is thought to help to reduce
blood cholesterol levels. Moreover, during the
present study AAq is believed to improve
coronary healthy by elevating high-density
lipoprotein levels (HDLs or "good"
cholesterol) in the blood, while decreasing the
low-density lipoproteins (LDLs, or "bad"
cholesterol). It may reduce the risk of
arteriosclerosis, stroke and heart attacks
(Gordon et al., 1989).
Phytochemical analysis has established
that leaves of Acacia tortilis contained a
variety of phenolic compounds (flavones,
flavonols,
flavonones,
anthocyanins,
leucoanthocyanins, coumarins and tannins)
and alkaloids (tertiary and quaternary
alkaloids)
in
varying
concentrations
(Nakafeero et al., 2007). In agreement with
our results, that ingestion of polyphenols of
red wine (Pal et al., 2003) and green tea
(Bursill et al., 2001) increase LDL-receptor
expression in a human hepatocyte cell line in
culture. In addition, they interfere with
solubilization of cholesterol in the digestive
tract of rats, thereby reducing cholesterol
absorption (Raederstorff et al., 2003) thus
Hypoglycemic and hypocholesterolemic effects of Acacia tortilis
6
decreasing the cholesterol and LDL level in
the present study.
In animals treated with AAq (Fig 2) a
decrease in body weight was also observed
which might be related to a decrease serum
cholesterol or glucose level. It is also reported
that Acacia tortilis contain flavanol (flavan-3ol) monomers (Nakafeero et al., 2007), have a
tendency to decreased body weight and waist
circumference (Chantre and Lairon, 2002).
Therefore the present study suggests that
administration of AAq (containing flavanol
catechins) inhibits the formation of oxidized
lipids such as LDL, and thus decreasing the
risk for developing arteriosclerosis. Moreover,
catechin intake decreases body fat. These
results suggest that catechins by decreasing the
body weight (Fig 2) contribute to the
prevention of diseases, particularly obesity.
The present study suggests that extracts of
Acacia tortilis leaves (AAq) possess significant
hypocholesterolemic
and
hypoglycemic
activity. Keeping this view in mind the
autopsy and toxicological studies were carried
out.
Oral administration of AAq (800mg/kg)
was found to be safe because no apparent
toxicity was observed under the experimental
conditions it did not cause lethality or organ
toxicity (liver, heart and kidney). Although to
confirm the toxicity of Acacia tortilis leaves
extract biochemical studies were carried out
showed that Acacia did not affect other blood
chemistry parameters. The non-significant data
collected after 7days were interpreted as
biological variability normally observed in
rats.
CONCLUSION
In conclusion, this study is unique;
showed that oral administered aqueous extract
(AAq at the dose of 800mg/kg) for 7 days only
causes
significant
hypoglycemia
and
hypolipidemia in normal rats. In the light of
present pharmacological and toxicological
studies, Acacia tortilis appears to be a valuable
plant, which can be useful, at least as an
adjunct, in the therapy of diabetes and hypercholesterolemia or in a condition in which
hyperglycemia and dyslipidemia coexist quite
often. Further the toxicological, hematological
and autopsy results further confirm the wide
marginal safety of Acacia tortilis leaves
extract.
ACKNOWLEDGEMENT
Authors are thankful to Miss Reham
Abdullah Badweelan, Miss Rehab Jameel AlHarbi, Miss Nehal Mohammad Sendi Faculty
of Applied Medical Sciences, Umm-Al-Qura
University, Makkah, Saudi Arabia, for
providing the research assistance.
REFERENCES
Alarcon-Aguilara, F.J., Roman-Ramos, R.,
Perez-Gutierrez, S., Aguilar-Contreras,
A., Contreras-Weber, C.C. and FloresSaenz, J.L. (1998). Study of the antihyperglycemic effect of plants used as
antidiabetics. J. Ethnopharmacol, 61: 101110.
Al-Nuaim, R.A.. (1997). High prevalence of
metabolic risk factors for cardiovascular
diseases among Saudi population, aged
30-64 years. Int. J. Cardiol., 62: 227-235.
Bursill, C., Roach, P.D., Bottema, C.D. and
Pal, S. (2001). Green tea upregulates the
low-density lipoprotein receptor through
the sterol-regulated element binding
protein in HepG2 liver cells. J. Agric.
Food Chem., 49: 5639-5645.
Chantre, P. and Lairon, D. (2002). Recent
findings of green tea extract AR25
(Exolise) and its activity for the treatment
of obesity. Phytomedicine, 9: 3-8.
Chopra, R.N., Nayar, S.L. and Chopra, I.C.
(1956). Acacia catechu Willd. Glossary of
Indian Medicinal plants. Council of
Scientific and Industrial Research. New
Delhi, India.
Waheeb Alharbi and Aisha Azmat
Durrington, P. (2003). Dyslipidaemia. Lancet
362(9385): 717-731.
Gordon, D.J., Probstfield, J.L., Garrison, R.J.,
Neaton, J.D., Castelli, W.P., Knoke, J.D.,
Jacobs, D.R., Bangdiwala, S. and Tyroler,
H.A.(1989). High-density lipoprotein
cholesterol and cardiovascular disease.
Four prospective American studies.
Circulation, 79(1): 8-15.
Haim, M., Benderly, M., Brunner, D., Behar,
S., Graff, E., Reicher-Reiss, H. and
Goldbourt, U. (1999). Elevated serum
triglyceride levels and long-term mortality
in patients with coronary heart disease.
Circulation, 100: 475-482.
Hokanson, J.E. and Austin, M.A. (1996).
Plasma triglycerides level is a risk factor
for cardiovascular disease independent of
high density lipoprotein cholesterol level:
meta-analysis of the population-based
prospective studies. J. Cardiovasc. Risk,
3: 213-219.
Iman, A. Hakima, Mohammed, A., Alsaifb,
Amal Aloudb, Mansour Alduwaihyc,
Khalid Al-Rubeaanc, Abdul Rahman AlNuaimc (2003). Black tea consumption
and serum lipid profiles in Saudi women:
a cross-sectional study in Saudi Arabia.
Nutrition Research, 23 1515-1526.
Jayasekhar, P. and Mohanan, P.V. (1997).
Rathinam K. Hepatoprotective activity of
ethyl acetate extract of Acacia catechu.
Indian J. Pharmacol., 29: 426-428.
Kirtikar, K.R. and Basu, B.D. (1935). Acacia
(Tourn.) Linn. In: Blatter, E., Caius, J.F.,
and Mhaskar, K., (editors). Indian
Medicinal Plants. 2nd ed. Dehradun:
International Book Distributors, Book
Sellers and Publishers, pp.919-935.
Meena, P.D, Kaushik, P., Shukla, S., Soni,
A.K., Kuma, M. and Kuma, A. (2006).
Anticancer and antimutagenic properties
of Acacia nilotica (Linn.) on 7,12dimethylbenz(a)anthracene-induced skin
papillomagenesis in Swiss albino mice.
Asian Pac. J. Cancer Prev., 7(4): 627632.
Metwally, M.A.A., El-Gellal A.M. and ElSawaisi. S.M. (2009). Effects of Silymarin
on Lipid Metabolism in Rats. World
7
Applied Sciences Journal, 6(12): 16341637.
Mohamed, S., Noordin, M.M., Baharuddin,
I.N., Abdullah, R. and Hamsan, K. (2002).
The effects of Cymbopogon citrates
(Serai) on the Heart, Liver and Kidney of
normal and hypercholesterolaemic rats. J.
Trop. Med. Plants, 3: 1-11.
Morozova, S., Suc-Royer, I. and Auwerx, J.
(2005).
Cholesterol
metabolism
modulators in future drug therapy for
atherosclerosis. Med Sci (Paris). 21:53-8.
Nagappa, A.N., Thakurdesai, P.A., Venkat
Rao, N. and Jiwan, S. (2003). Antidiabetic
activity of Terminalia catappa Linn fruits.
J. Ethnopharmacol., 88: 45-50.
Nakafeero, A.L., Mark, S. Reed and Nkbobi
M. Moleele (2007). Allelopathic potential
of five agroforestry trees, Botswana . Afr.
J. Ecol. 45:590-593.
Pal, S., Ho, N. and Santos, C. (2003). Red
wine polyphenolics increase LDL receptor
expression and activity and suppress the
secretion of ApoB100 from human
HepG2 cells. J. Nutr., 133: 700-7706.
Raederstorff, D.G., Schlachter, M.F., Elste, V.
and Weber, P. (2003). Effect of EGCG on
lipid absorption and plasma lipid levels in
rats. J. Nutr. Biochem., 14: 326-332.
Singh, K.N., Mettal, R.K. and Barthwal, K.C.
(1976). Hypoglycemia activity of Acacia
catechu, Acacia suma, and Albizzia
Odoratisima seed diets in normal albino
rats. Indian J. Med. Res., 64(5): 754-757.
Snow, V., Aronson, M., Hornbake, E., MotturPilson, C. and Weiss, K. (2004). Lipid
control in the management of type 2
diabetes mellitus: a clinical practice
guideline from the American College of
Physicians. Ann. Intern. Med., 140(8):
644-649.
Stacey, C. and Michael, C. (2007). The
complete herbal guide. A natural approach
to healing the body. Pub Lulu.com USA,
pp.173-175.
Tang, J.L., Armitage, J.M., Lancaster, T.,
Silagy, C.A., Fowler, G.H. and Neil, H.A.
(1998). Systematic review of dietary
intervention trials to lower blood total
8
Hypoglycemic and hypocholesterolemic effects of Acacia tortilis
cholesterol in free-living subjects. BMJ,
316(7139): 1213-1220.
Usher, G. (1984). Acacia catechu Willd
(Catechu, Dark catechu). A dictionary of
plant. 1st ed., CBS publishers and
distributors, Delhi, India.
Wallis, T.E. (1967). Cutch. Text book of
Pharmacognosy. 5th ed. J. and A.
Churchill Ltd., London.
WHO/Acadia (1992). Rapport de la Journe
Internationale de, diabetes.