6
BRIEF RESUME OF INTENDED WORK
6.1
NEED FOR THE STUDY
Hypercholesterolemia is the presence of high levels of cholesterol in the blood. It is not a disease
but a metabolic derangement that can be secondary to many diseases and can contribute to many
forms of disease, most notably cardiovascular disease. It is closely related to the terms
"hyperlipidemia" (elevated levels of lipids) and "hyperlipoproteinemia" (elevated levels of
lipoproteins)1.
Elevated cholesterol does not lead to specific symptoms unless it has been longstanding. Some
types of hypercholesterolemia lead to specific physical findings: xanthoma (thickening of tendons
due to accumulation of cholesterol), xanthelasma palpabrum (yellowish patches around the
eyelids) and arcus senilis (white discoloration of the peripheral cornea).
Longstanding elevated hypercholesterolemia leads to accelerated atherosclerosis; this can express
itself in a number of cardiovascular diseases1.
Amaranthus caudatus(Amaranthaceae) is a species of annual flowering plant. Many parts of the
plants, including the leaves and seeds, are edible, and are frequently used as a source of food in
India and South America. The mild flavored leaves are rich in vitamins and minerals2.
Plant used for purifying blood and in piles and as diuretic in stranguary also given in scrofula and
applied to scofullous sores3.
Amaranthus caudatus is used as diuretic and astringent 4, it is used as anthelmintic and the as
mild flavored leaves are rich in vitamins and minerals5. It is used as vegetable for enriching iron
deficiency6.
Nutritionally amaranth grain has 2-5 times higher biological value than a common cereal grain,
while containing substantially higher levels of proteins with 2-3 times higher lysine content.
Current industrial and public interest in the use of amaranth grain has not only linked to its
recognized nutritional properties, but also to its potential beneficial use as therapeutic adjuvant in
diets, hypercholesterolemic susceptible individuals. Recent studies have demonstrated a variety of
important and unique nutraceutical type applications for grain amaranth, including its use as an
adjunct to lower glucose response in non insulin dependent diabetics and in other application to
lower blood serum cholesterol level 7. The reports have shown that Amaranthus grain lowers the
blood serum cholesterol level, hence an attempt will be made to investigate leaves of Amaranthus
caudatus for the anti cholesterolemic activity.
1
6.2
REVIEW OF LITERATURE
● Filomena conforti .et. al., (2005) have reported in Vitro antioxidant effect and inhibition
of alpha-amylase of two varieties of Amaranthus caudatus seeds7.
● Klimczak I, Małecka M, Pachołek B (2002) have reported Antioxidant activity of ethanolic
extracts of amaranth seeds8.
.
● Andrea Y. A. Plate and José A. G. Areas (2002) have reported Cholesterol-lowering effect of
extruded amaranth (Amaranthus caudatus L.) in hypercholesterolemic rabbits9.
● Rinderle SJ, Goldstein IJ, Matta KL, Ratcliffe RM (1989) have reported, Isolation and
characterization of amaranthin, a lectin present in the seeds of Amaranthus caudatus, that
recognizes the T- (or cryptic T)-antigen10.
● Hejgaard J, Dam J, Petersen LC, Bjørn SE have reported primary structure and specificity of the
major serine proteinase inhibitor of amaranth (Amaranthus caudatus L.) seeds11.
● Rinderle SJ, Goldstein IJ, Remsen EE (1990) have reported physicochemical properties of
amaranthin, the lectin from Amaranthus caudatus seeds12.
● Pedersen B, Knudsen KE, Eggum BO (1990) have reported the nutritive value of amaranth
grain (Amaranthus caudatus) , energy and fibre of raw and processed grain13.
● Bressani R, Kalinowski LS, Ortiz MA, Elías LG (1987) have reported nutritional evaluation
of roasted, flaked and popped A. caudatus14.
● Bunzel M, Ralph J, Steinhart H (2005) have reported association of non-starch
Polysaccharides and ferulic acid in grain amaranth (Amaranthus caudatus L.) dietary
fibre15.
2
● Bruni R, Guerrini A, Scalia S, Romagnoli C, Sacchetti G(2002) have reported rapid
techniques for the extraction of vitamin E isomers from Amaranthus caudatus seeds:
ultrasonic and supercritical fluid extraction16.
● Zeng F. et. al.,(2003) have reported the effect of rare earth element europium on amaranthin
synthesis in Amarathus caudatus seedlings17.
● Bruni R. et. al.,(2001) have reported the wild Amaranthus caudatus seed oil, a nutraceutical
resource from Ecuadorian flora 18.
● Bouyoussfi M, Laus G, Verheyden P, Wyns L, Tourwe D, Van Binst G(1997) have reported
location of the three disulfide bonds in an antimicrobial peptide from Amaranthus caudatus
using mass spectrometry19.
● De Bolle MF. et. al.,(1996) have reported antimicrobial peptides from Mirabilis jalapa and
Amaranthus caudatus: expression, processing, localization and biological activity in
transgenic tobacco20.
● Verheyden P. et. al.,(1995) have reported the NMR study of the interaction of N,N',N"triacetyl chitotriose with Ac-AMP2, a sugar binding antimicrobial protein isolated from
Amaranthus caudatus21.
● De Bolle MF, David KM, Rees SB, Vanderleyden J, Cammue BP, Broekaert WF(1993) have
reported the cloning and characterization of a cDNA encoding an antimicrobial chitin-binding
protein from amaranth, Amaranthus caudatus22.
● Broekaert WF. et. al.,(1992) have reported the antimicrobial peptides from Amaranthus
caudatus seeds with sequence homology to the cysteine/glycine-rich domain of chitinbinding proteins24.
3
6.3
OBJECTIVES OF THE STUDY
a)
b)
c)
d)
Collection and authentication of Amaranthus caudatus Leaves.
Priliminary extraction of the drug using different solvents.
Phytochemical studies of the extracts.
Evaluation of anticholesterolemic activity.
7.0
MATERIAL AND METHODS
7.1
SOURCE OF DATA
Search on Pubmed , Science direct and other journals. RGUHS- Digital library,
Drug discovery and evaluation, Pharmacological Assays ,Second Edition by H.Gerhard Vogel
Place of Work: P. E. S. College of Pharmacy, Bengalooru -50
7.2
METHOD OF DATA COLLECTION
PLAN OF WORK
Collection of Plants: Leaves of the plant will be collected from G.K.V.K., Bengalooru and
authenticated by the Department of Botany, PES Degree College, Bengalooru.
Preliminary extraction: The extracts will be subjected to thorough Phytochemical studies.
Evaluation of anti-cholesterolemic activity: The different extracts will be further
evaluated for anti-cholesterolemic activity in rats.
Triton induced anti-cholesterolemic activity.
Hypocholesterolemic activity in rats.
7.3
DOES THE STUDY REQUIRE ANY INVESTIGATION TO BE CONDUCTED ON
PATIENTS OR ANIMALS?
-Yes-
7.4
HAS ETHICAL CLEARANCE BEEN OBTAINED FROM YOUR INSTITUTION IN
7.3 ?
Yes - Ethical clearance been obtained from IAEC
4
8.
Bibliography:1. http.//en.wikipedia.org/wiki/Hypercholesterolemia.[cited 2007 Nov 6th].
2. http://www.underutilized-species.org/species/species_details.asp id=42 . [cited 2007 Nov 6th].
3. Chopra.R.N, Nayar.S.L and Chopra.I.C. Glossary of Indian medicinal plants .1980; P 15 .
4. Evans.W.C. , Pharmacognosy , 15th ed. P 471 .
5. Yoganarasimhan.S.N. Medicinal plants of India Tamilnadu . Vol 2; 2000; P38.
6. http://www.pfaf.org/database/plants.php Amaranthus+caudatus .[cited 2007 Nov 6th].
7. Filomena conforti ,Giancarlo statti ,Monica Rosa Loizzo ,Gianni Sacchetti, Ferruccio Poli
and Francesco Menichini .In Vitro antioxidant effect and inhibition of alpha-amylase of two
varieties of Amaranthus caudatus seeds. Biol Pharm Bull. 2005 Jun; 28 (6):1098-102.
8. Klimczak I, Małecka M, Pachołek B. Antioxidant activity of ethanolic extracts of amaranth
seeds. Nahrung. 2002 Jun; 46(3):184-6.
9. Andrea Y. A. Plate and José A. G. Arêas Cholesterol-lowering effect of extruded amaranth
(Amaranthus caudatus L.) in hypercholesterolemic rabbits Food Chemistry, Volume 76, Issue
1, January 2002, Pages 1-6.
10.Rinderle SJ, Goldstein IJ, Matta KL, Ratcliffe RM. Isolation and characterization of
amaranthin, a lectin present in the seeds of Amaranthus caudatus, that recognizes the T- (or
cryptic T)-antigen. J Biol Chem. 1989 Sep 25; 264 (27):16123-31.
11. Hejgaard J, Dam J, Petersen LC, Bjørn SE. Primary structure and specificity of the major
serine proteinase inhibitor of amaranth (Amaranthus caudatus L.) seeds. Biochim Biophys Acta.
12. Rinderle SJ, Goldstein IJ, Remsen EE. Physicochemical properties of amaranthin, the lectin
from Amaranthus caudatus seeds. Biochemistry. 1990 Nov 20; 29 (46):10555-61.
13. Pedersen B, Knudsen KE, Eggum BO. The nutritive value of amaranth grain (Amaranthus
caudatus). Energy and fibre of raw and processed grain. Plant Foods Hum Nutr. 1990
Jan; 40 (1):61-71.
14. Bressani R, Kalinowski LS, Ortiz MA, Elías LG. Nutritional evaluation of roasted, flaked
and popped A. caudatus. Arch Latinoam Nutr. 1987 Sep;37(3):525-31.
15. Bunzel M, Ralph J, Steinhart H. Association of non-starch polysaccharides and ferulic acid
in grain amaranth (Amaranthus caudatus L.) dietary fiber.Mol Nutr Food Res. 2005
Jun; 49(6):551-9.
5
16.Bruni R, Guerrini A, Scalia S, Romagnoli C, Sacchetti G. Rapid techniques for the extraction
of vitamin E isomers from Amaranthus caudatus seeds: ultrasonic and supercritical fluid
extraction. Phytochem Anal. 2002 Sep-Oct; 13(5):257-61.
17.Zeng F, Tian HE, Wang Z, An Y, Gao F, Zhang L, Li F, Shan L. Effect of rare earth element
europium on amaranthin synthesis in Amarathus caudatus seedlings. Biol Trace Elem Res.
2003 Summer; 93(1-3):271-82.
18.Bruni R, Medici A, Guerrini A, Scalia S, Poli F, Muzzoli M, Sacchetti G. Wild Amaranthus
caudatus seed oil, a nutraceutical resource from Ecuadorian flora. J Agric Food Chem. 2001
Nov;49(11):5455-60.
19. Bouyoussfi M, Laus G, Verheyden P, Wyns L, Tourwe D, Van Binst G. Location of the
three disulfide bonds in an antimicrobial peptide from Amaranthus caudatus using mass
spectrometry. J Pept Res. 1997 Apr; 49(4):336-40.
20. De Bolle MF, Osborn RW, Goderis IJ, Noe L, Acland D, Hart CA, Torrekens S, Van Leuven
Broekaert WF. Antimicrobial peptides from Mirabilis jalapa and Amaranthus caudatus:
expression, processing, localization and biological activity in transgenic tobacco. Plant Mol
Biol. 1996 Aug; 31(5):993-1008.
21.Verheyden P, Pletinckx J, Maes D, Pepermans HA, Wyns L, Willem R, Martins JC. 1H
NMR study of the interaction of N,N',N"-triacetyl chitotriose with Ac-AMP2, a sugar binding
antimicrobial protein isolated from Amaranthus caudatus. FEBS Lett. 1995 Aug
21; 370 (3):245-9.
22. De Bolle MF, David KM, Rees SB, Vanderleyden J, Cammue BP, Broekaert WF. Cloning
and characterization of a cDNA encoding an antimicrobial chitin-binding protein from
amaranth, Amaranthus caudatus. Plant Mol Biol. 1993 Sep;22(6):1187-90.
23. H.Gerhard Vogel(Ed.). Drug discovery and evaluation, Pharmacological assays. 2nd ed.
P 1103, 1104,1106, 1107.
24. Broekaert WF, Mariën W, Terras FR, De Bolle MF, Proost P, Van Damme J, Dillen L,
Claeys M, Rees SB, Vanderleyden J, et al. Antimicrobial peptides from Amaranthus
caudatus seeds with sequence homology to the cysteine/glycine-rich domain of chitinbinding proteins. Biochemistry. 1992 May 5; 31(17):4308-14.
6