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Acta Poloniae Pharmaceutica ñ Drug Research, Vol. 68 No. 2 pp. 291ñ294, 2011 HYPOLIPIDEMIC EFFECT OF FRESH TRITICUM AESTIVUM (Wheat) GRASS 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
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
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, Triticum aestivum 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 tivum 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). Low
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 Phytochemical screening
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
Statistical analysis
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 (wheat) grass.
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 < 0.05).
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).
Triticum aestivum 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 grass 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 GJ. Bioactive 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 tivum 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, tivum 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- REFERENCES
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The authors wish to express their gratitude to Dr.R.Vijayraghvan Director and Mr. Dev Kumar

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