Acta Poloniae Pharmaceutica ñ Drug Research, Vol. 68 No. 2 pp. 291ñ294, 2011 ISSN 0001-6837
Polish Pharmaceutical Society
Cardiovascular diseases are the most common
cause of death worldwide. Abnormalities in plasma
lipoprotein and derangement in lipid metabolism
rank as the most firmly established and best under-
stood risk factor for atherosclerosis and cardiovas-
cular complications (1). Approximately 10% of the
global population is affected by dyslipidemia (2).
Therapeutic approaches for prevention of athero-
sclerosis are largely based on the use of statins,
which inhibit the rate limitng enzyme of cholesterol
biosynthesis. Currently inhibition of intestinal cho-
lesterol absorption by interfering with the sterol
transporting system is reported as novel mechanism
for lowering of serum cholesterol (3). This mecha-
nism is complimentary to that of statins, which
decrease the endogenous synthesis of cholestrerol.
Statins are found less beneficial in persons where
LDL receptors are low as in homozygous hypercho-
lesterolemia (4). Myopathy is the most troublesome
adverse effect of statins (5). Importance of natural
products in modern medicine is increased recently.
Natural products are important source of new drugs
and lead compounds, suitable for further modifica-
tion during drug development (6). Triticum aestivum
L. (family: Poaceae) grass commonly known as
wheat grass, is the freshly sprouted shoot of grain
wheat that has been used as herbal medicine in pres-
ent and past cultures and is highly valued for its
therapeutic and nutritional properties but lacking
scientific validation. Lipid lowering effect of fresh
Triticum aestivum grass juice (GJ) in normal rats is
reported (7). The present study is aimed to assess
hypolipidemic potential of fresh Triticum aestivum
grass juice in hypercholesterolemic rats. Effect of
grass juice on fecal cholesterol excretion was evalu-
ated to investigate mechanism of hypolipidemic
MATERIALS AND METHODS
Growing of the grass
The grass of Triticum aestivum used in this
study was grown indoors until required for experi-
ments. Earthen pot was filled with 2.5 inches of
HYPOLIPIDEMIC EFFECT OF FRESH TRITICUM AESTIVUM (Wheat) GRASS
JUICE IN HYPERCHOLESTEROLEMIC RATS
SAROJ KOTHARI*, ANAND K. JAIN, SWAROOP C. MEHTA and SHRINIVAS D. TONPAY
Department of Pharmacology, Gajara Raja Medical College, Gwalior, M.P., India
Abstract: Present study was aimed to elucidate hypolipidemic effect of fresh Triticum aestivum (common
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
aestivum 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 GJ
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.
Keywords: Triticum aestivum, grass, hypercholesterolemia, atorvastatin
* Corresponding author: e-mail: saroj.kothari@ rediff.com; phone-91-751-4082868; Mobile 9827322002
SAROJ KOTHARI et al.
growing medium composed of 3 parts of soil and
one part of compost. Overnight soaked Triticum aes-
tivum seeds were then evenly spread over it and fur-
ther covered with 0.5 inch of soil. Small quantities
of water were sprinkled evenly over soil and 3ñ4 h
indirect sunlight was allowed daily for growth of
grass. On the tenth day, when grass is about 6 inch-
es tall, it is cut 0.5 inch above the surface of soil. To
harvest continuous supply of fresh grass, pots were
similarly planted at one-day interval (7).
Preparation of fresh grass juice
Twenty grams of above harvested fresh grass
was grounded in a laboratory mortar and the juice
was squeezed out through four layers of wet muslin
cloth. The residue was twice resuspended in 3 mL of
sterile water and similarly squeezed. The filtrate was
made up to 20 mL (w/v) final volume with sterile
water and administered as GJ. Each day the fresh
juice was prepared prior to administration (7).
Randomly bred six to eight weeks old Wistar
rats of both sexes, weighing 150ñ200 g, raised in the
animal house of the Department of Pharmacology,
Gajara Raja Medical College, Gwalior, (M.P.),
India, were used for the study. These were main-
tained at 24 ± 2OC with 12 h light and dark cycle and
kept on standard pellet diet (Pranav Agro Industries
Delhi, India) and water ad libitum. The care and
maintenance of animals was according to the
approved guidelines of the Committee for the
Purpose of Control and Supervision of Experiments
on Animals (CPSCEA) in India. The Institutional
Animal Ethics Committee approved the protocol.
Induction of hypercholesterolemia
Hypercholesterolemia was induced experimen-
tally in rats by including 0.75 g% cholesterol and 1.5
g% bile acid in normal diet for 14 days (8).
Protocol for antihyperlipidemic activity
The experimental animals were divided into
five groups of six animals in each group and
received following treatments for 14 days by gav-
The first group (NDB), served as baseline
parameter and was treated with normal diet. The
second group (HCG) served as hypercholes-
terolemic control and was administered with high
cholesterol diet. The third, fourth and fifth group
(HTA5, HTA10 and HSG) animals were given grass
juice at the dose of 5 mL/kg, 10 mL/kg and standard
hypolipidemic drug atorvastatin 0.02% w/v in 2%
gum acacia suspension at a dose of 1 mg/kg, respec-
tively, along with high cholesterol diet. All the
above treatments were carried out each day in the
morning under similar constant conditions, as far as
Lipid profile measurements
After 24 h of the last dose administration the
animals were anesthetized with diethyl ether and
blood samples were collected by orbital puncture.
These were allowed to clot and then were cen-
trifuged at 3000 rpm/10 min and serum was used for
estimation of total cholesterol (TG) by CHOD Pap,
triglycerides (TG) by GPO Pap and high density
lipoprotein-cholesterol (HDL-C) by direct method
using commercial enzymatic kits (Randox, UK) and
Photometer model BTR-830 (Biotech, Spain). Low
density lipoprotein-cholesterol (LDL-C) and very
low density lipoprotein-cholesterol (VLDL-C) were
calculated using Friedewaldís formula (9).
Fecal cholesterol excretion
Fecal matter was collected during last 3 days of
the treatment period. The dried and powdered fecal
matter was extracted with chloroform : methanol
(2:1, v/v) mixture. The resultant extract was then
analyzed for cholesterol content in a manner similar
to that of serum. Cholesterol excreted in fecal mat-
ter (mg/g) was calculated (10).
Fresh Triticum aestivum GJ was subjected to
phytochemical tests for presence of bioactive com-
pounds by standard methods as described by
Statistical evaluation was done using one-way
ANOVA followed by Student-Newman-Keulís
multiple comparison tests. Differences with p < 0.05
were considered significant. Data are presented as
the mean ± SD. All statistical analyses were per-
formed by Sigma Stat software version 2.0, Jandel
Scientific Inc. USA.
Supplementation of high cholesterol diet in rats
(HSG) for 14 days resulted in significant (p < 0.05)
increase in TC and LDL-C levels as compared to the
rats treated with normal diet (NDC). TC and LDL-C
levels were increased by 202% and 333%, respec-
tively (Table 1). The levels of TC, TG, LDL-C and
VLDL-C were decreased by 50, 22, 56 and 22 per-
Hypolipidemic effect of fresh Triticum aestivum (wheat) grass...
cent in rats treated with grass juice at the dose of 5
mL/kg (HTA5) and by 60, 38, 69 and 38 percent in
rats 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 < 0.05).
HDL-cholesterols shown an increase by 12 and 15
percent in HTA5 and HTA10, respectively, and
were not significant (p > 0.05). Administration of
standard drug, atorvastatin, resulted in a decrease of
TC, TG, LDL-C, and VLDL-C by 66, 40, 72 and
40%, respectively, in HSG group and was signifi-
cant (p < 0.05). HDL-C levels showed 6% non sig-
nificant increase (p > 0.05).
Triticum aestivum grass juice induced 28 and
50% increase in cholesterol excretion at 5 mL/kg
and 10 mL/kg doses, respectively, in fecal matter.
The differences in cholesterol excretion were signif-
icant (p < 0.05) at the two test doses of GJ (Figure
Phytochemical tests revealed the presence of
flavonoids, triterpenoids, anthranol, alkaloids, tan-
nins, saponins. and sterols in fresh grass juice.
High cholesterol diet-induced experimental
hypercholesterolemic rat model has been used to
study hypolipidemic effects of plant extract (8). The
results of present study clearly show that oral fresh
grass juice of Triticum aestivum has dose dependent
significant hypolipidemic activity on diet-induced
raised levels of TC, TG, LDL-C and VLDL-C as
compared to control. Lipid levels at the dose of 10
mL /kg were comparable with that of standard drug ñ
atorvastatin. These changes in lipid levels after grass
juice treatment may be attributed to bioactive com-
pounds that were demonstrated after phytochemical
screening of fresh Triticum aestivum GJ. Bioactive
plant compounds ñ flavonoids and triterpenoids, are
reported to modulate lipid levels (12, 13). The pres-
ence of flavonoids and triterpenoids in Triticum aes-
tivum grass juice might have contributed in lipid low-
ering effect of Triticum aestivum grass juice in similar
manner. Tannins are reported to increase in activity of
the endothelium bound lipoprotein lipase activity,
Table 1. Effect of fresh grass juice of Triticum aestivum and atorvastatin on serum lipid profile of hypercholesterolemic rats.
Very low density
Group Total cholesterolTriglycerides
NDB 69.25 ± 3.62 58.31 ± 4.80 14.36 ± 1.11 43.14 ± 3.63 11.66 ± 0.96
HCG 209.83 ± 14.66a
63.63 ± 11.93 10.21 ± 1.57 186.89 ± 17.56 12.72 ± 2.38
HTA5 104.21 ± 5.31ab
49.76 ± 4.89a
11.45 ± 1.04 82.81 ± 3.44a
9.95 ± 0.97a
HTA10 76.18 ± 4.87ab
39.50 ± 3.91a
11.75 ± 0.79 58.56 ± 4.10ab
7.89 ± 0.78a
HSG 72.26 ± 3.94ab
38.29 ± 7.86a
11.90 ± 0.85 52.74 ± 3.84ab
Values are the mean ± SD, n = 6 in each group. The lipid profiles are given in mg/dL * p < 0.05 (Student-Newmen-Keulís multiple
comparison 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 dose
of 5 mL/kg. HTA10 = rats treated with high cholesterol diet and Triticum aestivum grass juice at the dose of 10 mL/kg. HSG = rats
treated with high cholesterol diet and atorvastatin at the dose of 1 mg/kg
Figure 1. Effect of fresh Triticum aestivum grass juice on fecal
cholesterol excretion in hypercholesterolemic rats. Each column
represents the mean ± SD, n = 6. *p < 0.05 as compared with con-
trol group. HSG = rats treated with high cholesterol diet. HTA5 =
rats treated with high cholesterol diet and Triticum aestivum grass
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
SAROJ KOTHARI et al.
which hydrolyzes triglycerides as reported by Tebib
et al. (14). The presence of tannins in Triticum aes-
tivum grass juice might be involved in triglyceride
lowering activity but this need to be investigated by
further studies. Cholesterol absorption inhibitors are
novel class of drugs reducing cholesterol levels and
ezetimibe is the first in this class (15). Cholesterol in
the intestine can arise both from the diet and hepatic
secretions. Further, inhibition of cholesterol absorp-
tion from intestine also decreases the delivery of cho-
lesterol to the liver, thereby lowering serum as well as
hepatic cholesterol. This, in turn, accelerates the
uptake of LDL from plasma viaLDL receptors and an
increase in the clearance of plasma cholesterol (16).
Plant sterols are also reported to decrease cholesterol
absorption but through a different mechanism.
Phytosterols compete with dietary and billiary cho-
lesterol for incorporation into mixed miscelles in the
intestinal lumen thus inhibiting their uptake (17). To
study the effect of fresh Triticum aestivum GJ on
intestinal absorption of cholesterol, fecal cholesterol
levels were measured. A dose dependent significant
increase in cholesterol content of fecal matter of
hypercholesterolemic rats after Triticum aestivum GJ
administration indicates interference in absorption of
intestinal cholesterol. It is suggested that sterols found
in grass juice might have decreased cholesterol
absorption and increased cholesterol excretion there-
by contributed in hypolipidemic activity of Triticum
aestivum. Saponins are another highly active plant
compounds reported to increase fecal cholesterol
excretion (18). It is tempting to suggest that besides
sterols, the presence of saponins in Triticum aestivum
might have contributed in increasing fecal cholesterol
excretion. The effect of fresh GJ on cholesterol syn-
thesis is not investigated in the present study and the
results found are encouraging to explore it in further
Thus, it is concluded that fresh Triticum aes-
tivum grass juice possess hypolipidemic activity in
hypercholesterolemic rats and it increases choles-
terol excretion. It might be useful with statins for the
management of dyslipidemia. This is the first study
which investigated the hypolipidemic activity of
Triticum aestivum grass juice in hypercholes-
terolemic rats and its effect on cholesterol excretion.
The results found are encouraging for further assess-
ment to elucidate other mechanisms of hypolipi-
The authors wish to express their gratitude to
Dr.R.Vijayraghvan Director and Mr. Dev Kumar
Development Establishment, Ministry of Defence,
Government of India, Gwalior, for their unstinted
help. We are also thankful to Dr. (Mrs.) Shaila
Sapre, Dean, Gajara Raja Medical College and J.A.
Group of Hospitals, Gwalior (M.P.), India for finan-
officer Defence, Research and
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Received: 23. 03. 2010