Acta Poloniae Pharmaceutica ñ Drug Research, Vol. 68 No. 2 pp. 291ñ294, 2011
HYPOLIPIDEMIC EFFECT OF FRESH TRITICUM AESTIVUM
SAROJ KOTHARI*, ANAND K. JAIN, SWAROOP C. MEHTA and SHRINIVAS D. TONPAY
Department of Pharmacology, Gajara Raja Medical College, Gwalior, M.P., India
Present study was aimed to elucidate hypolipidemic effect of fresh Triticum aestivum
wheat) grass juice (GJ) in experimentally induced hypercholesterolemia in rats and to investigate its role in cho-
lesterol excretion. Hypercholesterolemia was induced experimentally in rats by including 0.75 g% cholesterol
and 1.5 g% bile salts in normal diet for 14 days. Hypercholesterolemic rats were administered fresh Triticum
GJ at the dose of 5 mL/kg and 10 mL/kg and the standard drug atorvastatin 0.02% w/v in 2% gum
acacia suspension at the dose of 1 mg/kg for 14 days by gavage. Blood samples were collected after 24 h of last
administration and used for estimation of lipid profile. Fecal cholesterol levels were estimated using standard
methods. Fresh GJ administration at 5 mL/kg and 10 mL/kg resulted in dose dependent significant decline in
total cholesterol (TC), triglycerides (TG), low density lipoprotein-cholesterol (LDL-C) and very low density
lipoprotein-cholesterol (VLDL-C) levels in hypercholesterolemic rats. Further, in comparison to atorvastatin,
GJ administration at the dose of 10 mL/kg resulted in comparable decrease of TC, LDL-C, TG and VLDL-C
levels (p > 0.05). Fecal cholesterol excretion was significantly (p < 0.05) enhanced by Triticum aestivum
administration. Phytochemical analysis revealed the presence of flavonoids, triterpenoids, anthraquinol, alka-
loids, tannins, saponins and sterols in fresh wheat grass juice. The results of present study revealed hypolipi-
demic effect of Triticum aestivum
GJ in hypercholesterolemic rats by increasing fecal cholesterol excretion.
Fresh GJ could have potentially beneficial effect in atherosclerosis associated with hyperlipidemia.
Triticum aestivum, grass, hypercholesterolemia, atorvastatin
Cardiovascular diseases are the most common
and lead compounds, suitable for further modifica-
cause of death worldwide. Abnormalities in plasma
tion during drug development (6). Triticum aestivum
lipoprotein and derangement in lipid metabolism
L. (family: Poaceae) grass commonly known as
rank as the most firmly established and best under-
wheat grass, is the freshly sprouted shoot of grain
stood risk factor for atherosclerosis and cardiovas-
wheat that has been used as herbal medicine in pres-
cular complications (1).
Approximately 10% of the
ent and past cultures and is highly valued for its
global population is affected by dyslipidemia (2).
therapeutic and nutritional properties but lacking
Therapeutic approaches for prevention of athero-
scientific validation. Lipid lowering effect of fresh
sclerosis are largely based on the use of statins,
grass juice (GJ) in normal rats is
which inhibit the rate limitng enzyme of cholesterol
reported (7). The present study is aimed to assess
biosynthesis. Currently inhibition of intestinal cho-
hypolipidemic potential of fresh Triticum aestivum
lesterol absorption by interfering with the sterol
grass juice in hypercholesterolemic rats. Effect of
transporting system is reported as novel mechanism
grass juice on fecal cholesterol excretion was evalu-
for lowering of serum cholesterol (3). This mecha-
ated to investigate mechanism of hypolipidemic
nism is complimentary to that of statins, which
decrease the endogenous synthesis of cholestrerol.
Statins are found less beneficial in persons where
MATERIALS AND METHODS
LDL receptors are low as in homozygous hypercho-lesterolemia (4). Myopathy is the most troublesome
Growing of the grass
adverse effect of statins (5). Importance of natural
The grass of Triticum aestivum
used in this
products in modern medicine is increased recently.
study was grown indoors until required for experi-
Natural products are important source of new drugs
ments. Earthen pot was filled with 2.5 inches of
* Corresponding author: e-mail: saroj.kothari@ rediff.com; phone-91-751-4082868; Mobile 9827322002
growing medium composed of 3 parts of soil and
gum acacia suspension at a dose of 1 mg/kg, respec-
one part of compost. Overnight soaked Triticum aes-
tively, along with high cholesterol diet. All the
seeds were then evenly spread over it and fur-
above treatments were carried out each day in the
ther covered with 0.5 inch of soil. Small quantities
morning under similar constant conditions, as far as
of water were sprinkled evenly over soil and 3ñ4 h
indirect sunlight was allowed daily for growth ofgrass. On the tenth day, when grass is about 6 inch-
Lipid profile measurements
es tall, it is cut 0.5 inch above the surface of soil. To
After 24 h of the last dose administration the
harvest continuous supply of fresh grass, pots were
animals were anesthetized with diethyl ether and
similarly planted at one-day interval (7).
blood samples were collected by orbital puncture.
These were allowed to clot and then were cen-
Preparation of fresh grass juice
trifuged at 3000 rpm/10 min and serum was used for
Twenty grams of above harvested fresh grass
estimation of total cholesterol (TG) by CHOD Pap,
was grounded in a laboratory mortar and the juice
triglycerides (TG) by GPO Pap and high density
was squeezed out through four layers of wet muslin
lipoprotein-cholesterol (HDL-C) by direct method
cloth. The residue was twice resuspended in 3 mL of
using commercial enzymatic kits (Randox, UK) and
sterile water and similarly squeezed. The filtrate was
Photometer model BTR-830 (Biotech, Spain).
made up to 20 mL (w/v) final volume with sterile
density lipoprotein-cholesterol (LDL-C) and very
water and administered as GJ. Each day the fresh
low density lipoprotein-cholesterol (VLDL-C) were
juice was prepared prior to administration (7).
calculated using Friedewaldís formula (9).
Fecal cholesterol excretion
Randomly bred six to eight weeks old Wistar
Fecal matter was collected during last 3 days of
rats of both sexes, weighing 150ñ200 g, raised in the
the treatment period. The dried and powdered fecal
animal house of the Department of Pharmacology,
matter was extracted with chloroform : methanol
Gajara Raja Medical College, Gwalior, (M.P.),
(2:1, v/v) mixture. The resultant extract was then
India, were used for the study. These were main-
analyzed for cholesterol content in a manner similar
tained at 24 ± 2OC with 12 h light and dark cycle and
to that of serum. Cholesterol excreted in fecal mat-
kept on standard pellet diet (Pranav Agro Industries
Delhi, India) and water ad libitum
. The care andmaintenance of animals was according to the
approved guidelines of the Committee for the
Fresh Triticum aestivum
GJ was subjected to
Purpose of Control and Supervision of Experiments
phytochemical tests for presence of bioactive com-
on Animals (CPSCEA) in India. The Institutional
pounds by standard methods as described by
Animal Ethics Committee approved the protocol.
Induction of hypercholesterolemia
Hypercholesterolemia was induced experimen-
Statistical evaluation was done using one-way
tally in rats by including 0.75 g% cholesterol and 1.5
g% bile acid in normal diet for 14 days (8).
multiple comparison tests. Differences with p < 0.05were considered significant. Data are presented as
Protocol for antihyperlipidemic activity
the mean ± SD. All statistical analyses were per-
The experimental animals were divided into
formed by Sigma Stat software version 2.0, Jandel
five groups of six animals in each group and
received following treatments for 14 days by gav-age.
The first group (NDB), served as baseline
parameter and was treated with normal diet. The
Supplementation of high cholesterol diet in rats
second group (HCG) served as hypercholes-
(HSG) for 14 days resulted in significant (p < 0.05)
terolemic control and was administered with high
increase in TC and LDL-C levels as compared to the
cholesterol diet. The third, fourth and fifth group
rats treated with normal diet (NDC). TC and LDL-C
(HTA5, HTA10 and HSG) animals were given grass
levels were increased by 202% and 333%, respec-
juice at the dose of 5 mL/kg, 10 mL/kg and standard
tively (Table 1). The levels of TC, TG, LDL-C and
hypolipidemic drug atorvastatin 0.02% w/v in 2%
VLDL-C were decreased by 50, 22, 56 and 22 per-
Hypolipidemic effect of fresh Triticum aestivum
Table 1. Effect of fresh grass juice of Triticum aestivum
and atorvastatin on serum lipid profile of hypercholesterolemic rats.
Values are the mean ± SD, n = 6 in each group. The lipid profiles are given in mg/dL * p < 0.05 (Student-Newmen-Keulís multiplecomparison test), a significant from control group (HCG), b significant from (HTA5) group; NDB = rats treated with normal diet, HSG= rats treated with high cholesterol diet. HTA5 = rats treated with high cholesterol diet and Triticum aestivum
grass juice at the doseof 5 mL/kg. HTA10 = rats treated with high cholesterol diet and Triticum aestivum
grass juice at the dose of 10 mL/kg. HSG = ratstreated with high cholesterol diet and atorvastatin at the dose of 1 mg/kg
cent in rats treated with grass juice at the dose of 5mL/kg (HTA5) and by 60, 38, 69 and 38 percent inrats treated with grass juice at the dose of 10 mL/kg(HTA10), respectively, in comparison with HCG .
The decrease in TC, TG, LDL-C and VLDL-C lev-els were dose dependent and significant (p <
HDL-cholesterols shown an increase by 12 and 15percent in HTA5 and HTA10, respectively, andwere not significant (p > 0.05). Administration ofstandard drug, atorvastatin, resulted in a decrease ofTC, TG, LDL-C, and VLDL-C by 66, 40, 72 and40%, respectively, in HSG group and was signifi-cant (p < 0.05). HDL-C levels showed 6% non sig-nificant increase (p > 0.05).
grass juice induced 28 and
Figure 1. Effect of fresh Triticum aestivum
grass juice on fecal
50% increase in cholesterol excretion at 5 mL/kg
cholesterol excretion in hypercholesterolemic rats. Each column
and 10 mL/kg doses, respectively, in fecal matter.
represents the mean ± SD, n = 6. *p < 0.05 as compared with con-
The differences in cholesterol excretion were signif-
trol group. HSG = rats treated with high cholesterol diet. HTA5 =rats treated with high cholesterol diet and Triticum aestivum
icant (p < 0.05) at the two test doses of GJ (Figure
juice at the dose of 5 mL/kg . HTA10 = rats treated with high cho-
lesterol diet and Triticum aestivum
grass juice at the dose of 10
Phytochemical tests revealed the presence of
flavonoids, triterpenoids, anthranol, alkaloids, tan-nins, saponins. and sterols in fresh grass juice.
atorvastatin. These changes in lipid levels after grassjuice treatment may be attributed to bioactive com-
High cholesterol diet-induced experimental
pounds that were demonstrated after phytochemical
hypercholesterolemic rat model has been used to
screening of fresh Triticum aestivum
study hypolipidemic effects of plant extract (8). The
plant compounds ñ flavonoids and triterpenoids, are
results of present study clearly show that oral fresh
reported to modulate lipid levels (12, 13). The pres-
grass juice of Triticum aestivum
has dose dependent
ence of flavonoids and triterpenoids in Triticum aes-
significant hypolipidemic activity on diet-induced
grass juice might have contributed in lipid low-
raised levels of TC, TG, LDL-C and VLDL-C as
ering effect of Triticum aestivum
grass juice in similar
compared to control. Lipid levels at the dose of 10
manner. Tannins are reported to increase in activity of
mL /kg were comparable with that of standard drug ñ
the endothelium bound lipoprotein lipase activity,
which hydrolyzes triglycerides as reported by Tebib
et al. (14). The presence of tannins in Triticum aes-
Development Establishment, Ministry of Defence,
grass juice might be involved in triglyceride
Government of India, Gwalior, for their unstinted
lowering activity but this need to be investigated by
help. We are also thankful to Dr. (Mrs.) Shaila
further studies. Cholesterol absorption inhibitors are
Sapre, Dean, Gajara Raja Medical College and J.A.
novel class of drugs reducing cholesterol levels and
Group of Hospitals, Gwalior (M.P.), India for finan-
ezetimibe is the first in this class (15). Cholesterol in
the intestine can arise both from the diet and hepaticsecretions. Further, inhibition of cholesterol absorp-
tion from intestine also decreases the delivery of cho-lesterol to the liver, thereby lowering serum as well as
1. Singh R.B., Mengi S.A., Xu Y.J., Arneja A.S.,
hepatic cholesterol. This, in turn, accelerates the
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uptake of LDL from plasma via
LDL receptors and an
2. Shilpha S., Bairy K.: Indian J. Physiol.
increase in the clearance of plasma cholesterol (16).
Plant sterols are also reported to decrease cholesterol
3. Davis H.R.: International Congress series 1262,
absorption but through a different mechanism.
Phytosterols compete with dietary and billiary cho-
4. Goodman and Gilmanís The Pharmacological
lesterol for incorporation into mixed miscelles in the
Basis of Therapeutics. 10th edn., Hardman J.G.,
intestinal lumen thus inhibiting their uptake (17). To
Limbird L.E. Eds., The McGraw Hill Co. Inc.,
study the effect of fresh Triticum aestivum
intestinal absorption of cholesterol, fecal cholesterol
5. Chapman J.M., Carrie A.: Arterioscler.
levels were measured. A dose dependent significant
Thromb. Vasc. Biol. 25, 2441( 2005).
increase in cholesterol content of fecal matter of
6. Chin Y.W., Balunas M.J., Chai H.B., Kinghonn
hypercholesterolemic rats after Triticum aestivum
administration indicates interference in absorption of
7. Kothari S., Jain A.K., Mehta S.C., Tonpay S.D.:
intestinal cholesterol. It is suggested that sterols found
Indian J. Pharmacol. 40, 235 (2008).
in grass juice might have decreased cholesterol
8. Visavadiya N.P., Narsinhacharya A.V.R.L.:
absorption and increased cholesterol excretion there-
Indian J. Pharmacol. 37, 376 (2005).
by contributed in hypolipidemic activity of Triticum
9. Friedewald W.T., Levy R.I., Fredrickson D.S.:
. Saponins are another highly active plant
compounds reported to increase fecal cholesterol
10. Souza T.D., Mengi S.A., Hassarajani S.,
excretion (18). It is tempting to suggest that besides
Chattopadhyay S.: Indian J. Pharmacol. 39,196
sterols, the presence of saponins in Triticum aestivum
might have contributed in increasing fecal cholesterol
11. Harborne J.B.: Phytochemical methods: A
excretion. The effect of fresh GJ on cholesterol syn-
guide to modern techniques of plant analysis. 2nd
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edn., Chapmann and Hall, London 1983.
results found are encouraging to explore it in further
12. Koshi A.S., Anita L., Vijayalakshmi N. R.:
Thus, it is concluded that fresh Triticum aes-
13. Zhang Q., Chang Z., Yang J., Wang Q.:
grass juice possess hypolipidemic activity in
hypercholesterolemic rats and it increases choles-
14. Tebib K., Besancon P., Rounanet J.M.: J. Nutr.
terol excretion. It might be useful with statins for the
management of dyslipidemia. This is the first study
15. Sudhop T., L¸tiohann D., Kodal A., Igel M.,
which investigated the hypolipidemic activity of
Tribble D.L., Shah S., Perevoskaya I., von
grass juice in hypercholes-
Bergmann K.: Circulation 106, 1943 (2002).
terolemic rats and its effect on cholesterol excretion.
16. Patel S.B.: Curr. Cardiol. Rep. 6, 439 (2004).
The results found are encouraging for further assess-
17. Brufau G., Canela M.A., Refecas M.: Nutr. Res.
ment to elucidate other mechanisms of hypolipi-
18. Matsui Y., Kumagai H., Masuda H.: Nihon
Shokuhin Kagaku Kô
gakkai (Food Sci.
The authors wish to express their gratitude to
Dr.R.Vijayraghvan Director and Mr. Dev Kumar
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