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24 ReferencesPharmacognostic and hypoglycemic studies of Achyranthus aspera L.
Abstract
In the present study, the ethanolic extract of Achyranthus aspera L. Leaves (EEA) were investigated for their pharmacognostic characteristics and hypoglycemic effects. The pharmacognosy of the plant was carried out using microscopic and macroscopic tools. Macroscopic studies revealed that the leaves were cauline, ramel, opposite, exstipulate, simple, sub-sessile, ovate, entire, acute, unicostate, reticulate, rough, coriaceous and hairy. The stem was erect, herbaceous, quadrangular, branched, solid, green and pubescent, while various internal parts were observed using powder drug for microscopic study. The hypoglycemic effect of the plant was studied in healthy normoglycemic rabbits. The EEA was tested in three doses (100, 150 and 200 mg/kg), and a dose dependent hypoglycemic effect was observed. The most significant hypoglycemic effect was observed against higher dose (200 mg/kg) which remained for 5 h while the hypoglycemic effect remained significant for 3 h against the lower dose. It was concluded that EEA has hypoglycemic properties and it is recommended for the treatment of diabetes mellitus II.
Vol. 5(7), pp. 127-131, July 2013
DOI: 10.5897/JPP2013-0271
ISSN 2141-2502 © 2013 Academic Journals
http://www.academicjournals.org/JPP
Journal of Pharmacognosy and
Phytotherapy
Full Length Research Paper
Pharmacognostic and hypoglycemic studies of
Achyranthus aspera L.
Ishtiaq Ahmad1, Muhammad Ibrar1, Barkatullah1, Naveed Muhammad2*, Zahir Muhammad1
and Niaz Ali1
1Department of Botany, University of Peshawar, Peshawar, KPK, Pakistan.
2Department of Pharmacy, Hazara University, Havelain Campus, Abbottabad, KPK, Pakistan.
Accepted 27 May, 2013
In the present study, the ethanolic extract of Achyranthus aspera L. Leaves (EEA) were investigated for
their pharmacognostic characteristics and hypoglycemic effects. The pharmacognosy of the plant was
carried out using microscopic and macroscopic tools. Macroscopic studies revealed that the leaves
were cauline, ramel, opposite, exstipulate, simple, sub-sessile, ovate, entire, acute, unicostate,
reticulate, rough, coriaceous and hairy. The stem was erect, herbaceous, quadrangular, branched,
solid, green and pubescent, while various internal parts were observed using powder drug for
microscopic study. The hypoglycemic effect of the plant was studied in healthy normoglycemic rabbits.
The EEA was tested in three doses (100, 150 and 200 mg/kg), and a dose dependent hypoglycemic
effect was observed. The most significant hypoglycemic effect was observed against higher dose (200
mg/kg) which remained for 5 h while the hypoglycemic effect remained significant for 3 h against the
lower dose. It was concluded that EEA has hypoglycemic properties and it is recommended for the
treatment of diabetes mellitus II.
Key words: Pharmacognostic, hypoglycemic, Achyranthus aspera L.
INTRODUCTION
Achyranthus aspera L. is a common perennial herb,
growing as a wild plant, in natural and cultivated habitats.
A. aspera L. has a branched tap root system. The stem is
herbaceous, quadrangular, branched, solid, green and
pubescent. Leaves are simple and exstipulate. A spike
type of recemose inflorescence occurs in this specimen.
Almost all parts (seed, root, stem and leaf) of A. aspera
L. possess medicinal value (Lans, 2007) but leaves have
greater importance in the traditional (unani) system of
health and herbal medicine formulation. Alcoholic decoc-
tions of A. aspera L. are mostly used as anti-inflammatory
(Girach Aminuddin and Khan, 1992) and infertility
inducing agents (Chakraborty et al., 2002; Shibeshi et al.,
2006). Furthermore, the plant possesses lavicidal
(Bagavan et al., 2008), antioxidant (Edwin et al., 2008),
anticancer (Chakraborty et al., 2002), anti- diarrheal and
antidysentric (Boily and Van Puyvelde
, 1986; Girach Aminuddin and Khan, 1992), antimicrobial
(Boily and Van Puyvelde, 1986) properties. The
hypoglycemic effect of methanolic and aqueous extract of
the whole plant of A. aspera has been reported (Akhtar
and Iqbal, 1991). The pharmacognostic profile and
hypoglycemic effect of the leaves of this plant was
studied in order to validate traditional uses.
MATERIALS AND METHODS
Plant
The study was conducted in the Department of Botany, University
*Corresponding author. E-mail. drnaveedrph@gmail.com
128 J. Pharmacognosy Phytother.
of Peshawar. Plants of A. aspera was collected from different
localities in the district of Charsadda, KPK, Pakistan. Plants were
identified by Dr. Muhammad Ibrar, Department of Botany, and
specimen was deposited there in the herbarium under voucher
number 7455/Bot. The whole plant was powdered for microscopic
study.
Preparation of extract
The ethanolic extract of the leaves was prepared using our previous
reported method (Barkatullah et al., 2011; Muhammad and Saeed,
2011; Muhammad et al., 2012b, c). Powder of leaves was
macerated in ethanol for 10 to 15 days. The extract was filtered and
then evaporated. The dry extract was stored in refrigerator for use.
Animals
Domesticated rabbits weighing 1 kg obtained from the district
Charsadda District, Pakistan were used. The animals were kept at
standard laboratory conditions. The animals were shifted from
animal house to the laboratory one hour prior to start the
experiment.
Pharmacognostic study
The pharmacognostic profile of the plant was reported using our
previous reported method (Muhammad et al., 2012; Ismail et al.,
2011). Vein islet number, vein termination number, palisade ratio,
stomatal number and stomatal indices were calculated from 1 mm2
pieces of leaf of A. aspera boiled in concentrated chloral hydrate
solution under high power compound microscope.
Macroscopical features
The following macroscopic characters of plant were noted: size,
shape, margins, apex, surface, color, odour, taste, nature, texture
of leaves, stem and root were also studied (Wallis, 1985; Evans
and Trease, 2002).
Powder drug Study
Powdered drug was studied using chloral hydrate and iodine
solution under compound microscope (Compoud microscope).
Organoleptic evaluation (color, odour, taste, fracture and touch) of
the powdered drug was done using five senses (Pharmacopoeia,
1986).
Phytochemical Study
Qualitative chemical identification tests
Various phytochemical tests were performed for detection of
various constituents preliminary, using well established procedures
(Trease and Evans, 1989; Muhammad and Saeed, 2011).
Hypoglycemic effects
The healthy rabbits were divided into five groups of six. Group I was
treated with normal saline (10 ml/kg) and acted as a control, Group
II was treated with the standard hypoglycemic drug, glibenclimide,
Group III, IV and V were treated with the ethanolic extract of
Achyranthus aspera (EEA) at doses of 100, 150 and 200 mg/kg,
respectively. Blood glucose level was noted at the start of the
experiment and then after 1, 2, 3, 4 and 5 h, respectively.
Glibenclimide was used as a standard hypoglycemic drug. Swab
soaked in methylated spirit was used as a disinfectant during
collection of blood. Blood was obtained by puncturing the veins in
the pinna of each animal by using a sharp lancet . A swab soaked
in methylated spirit was used to disinfect the lancet after each use.
RESULTS
Macroscopical features
Leaves of A. aspera L. are cauline, ramel, opposite,
exstipulate, simple, sub-sessile, ovate, entire, acute,
unicostate, reticulate, rough, coriaceous and hairy. The
stem is erect, herbaceous, quadrangular, branched, solid,
green and pubescent. A tap and branched root system is
present.
Microscopical features
Vein islet No., vein termination No., palisade ratio,
stomatal No., and stomatal index are given as in Table 1.
Powder drug study
Trichomes of various types that is, multicellular, shagay,
uniciniate, uniseriate etc (a-d), glandular hairs (e-f),
tracheids (l), parenchyma cells (j), mesophyll cells (i),
fragments of small veins (h), starch granules (g),
fragments of epidermis and stomata (k) were found in the
powder drug. Organoleptic evaluation of the powder drug
is as seen in Figure 1.
Phytochemical screening tests
Alkaloid, saponin, lignin, ca-oxalate, starch, protein,
mucilage, tannin were identified in the plant extract.
(Table 2).
Hypoglycemic effect
The hypoglycemic effects of ethanolic extracts of A.
aspera L. was compared with standard (glibenclamide).
Both standard and ethanolic plant extracts reduced the
blood glucose level of rabbits significantly (P < 0.05). The
result showed that ethanolic plant extracts of A. aspera L.
possess strong hypoglycemic effects. Higher doses of
plant extract is more effective in decreasing blood
glucose level when compared to low doses. The efficacy
of low doses last for 2 to 3 h while high doses last for
more than 3 h. (Table 3).
Ahmad et al. 129
a
b
c
d
e
f
g
h
i
j
k
l
Figure 1. Powder drug study of the A. aspera. Multicellular (a), shagy (b), uniciniate
(c), uniseriate (d), glandular hairs (e-f), starch granules (g), fragments of small veins
(h), mesophyll cells (i), parenchyma cells (j), fragments of epidermis and stomata
(k) and tracheids (l).
Table 1. Organoleptic evaluation of A. aspera leaves.
S/No
Parameter
Observation
1
Color
Dark green
2
Odour
Pungent
3
Taste
Slightly disagreeable
4
Fracture
Fibrous
5
Touch
Soapy
DISCUSSION
The pharmacognostic profile is essential for crude drugs
as it helps in the identification of specific drugs within the
same genus. There are several plants within the same
genus with similar structure and with different micro-
scopic characteristics, however the medicinal value of
these morphological similar plants vary from each other.
Pharmacognostic studies help in the identification of
specific plants possessing organoleptic and microscopic
characteristics (Ismail et al., 2011).
Glibenclamide is a sulfonylurea derivative and
commonly used in the management of diabetes mellitus
type II; it causes hypoglycemia by stimulating beta cells
of pancreas and increasing release of insulin and
inhibiting glucagon secretion. As these effects require a
functional pancreas, it can lower blood sugar levels in
non-diabetic subjects (Soon and Tan, 2002) and a
similarity was obtained in our tested ethanolic extract. It is
very interesting to note that the ethanolic extract of the
leaves of our selected plant is used in the tradition
system of medicines in the management of various
ailments. Our results show that the extract can decrease
the serum glucose level in normoglycemic individuals.
However, an adverse effect is that the plant can decrease
the normal glucose level, just like glibenclamide,
therefore, it is recommended that in the use of this plant
as an anti-inflammatory or even for diabetic manage-
ment, the glucose level should be monitored periodically.
It is also worth mentioning that the plant should not be
used in high doses for the treatment of ailments other
than diabetes because it lowers the normal serum
glucose level in normal patients.
In diabetes, oxidative stress has been found to be
mainly due to an increased production of oxygen free
radicals and a sharp reduction of anti-oxidant defenses
(Oberley, 1988; Brownlee, 2001; Muhammad et al.,
2012a). Hence, compounds with both hypoglycemic and
anti-oxidative properties would be useful anti-diabetic
agents (Brownlee, 2001). It has been reported that the
methanolic extract of A. aspera is a good antidiabetic and
antioxidant (Akhtar and Iqbal, 1991; Edwin et al., 2008)
agent. The phytochemical study revealed the presence of
130 J. Pharmacognosy Phytother.
Table 2. Phytochemical screening of the EEA.
Phytochemicals
Result
Remarks
Alkaloid
Reddish brown
++
Saponin
Frothing
++
Lignin
Pink coloration
++
Ca-oxalate
Dissolve in HCl without effervescence
++
Starch
Deep blue coloration
++
Protein
Yellow ppt.
++
Mucilage
Gummy suspension
++
Tannin
Greenish coloration
++
Table 3. Effect EEA on blood glucose level of normoglycemic rabbits.
Treatments (mg/kg)
0 h
1 h
2 h
3 h
4 h
5 h
Control
153±2
151±1.5
152±2
149±2
147.7±2.5
142.3±1.15
Glb (0.2)
153.7±1.5
111.3±2.1**
113.6±1.5**
117.3±2.5**
120.3±1.5**
122.7±1.5**
EEA (100)
149.7±1.5
114.3±2.1**
118±2**
123.6±2.5**
133.7±3
139±2.6
EEA (150)
144±2
109.7±2.5**
127.7±2.1**
131.3±2.1**
133±2
135.3±2.1
EEA (200)
150.7±2.5
90.6±2.1**
101. 0±1.5**
111.3±2.5**
116.7±1.5**
124.7±1.5**
Glb= Glibenclamide, Data presented as mean ± SEM (n = 6). *P < 0.05, **P < 0.01.
starch which produce hyperglycemia but the presence of
alkaloids, saponin and tannin in our results as hypogly-
cemic. Besides A. aspera L, many other plants like
Averrhoa bilimbi (Pushparaj et al., 2000) and Indigofera
pulchra (Tanko et al., 2008) have hypoglycemic effects
on blood glucose level of normoglcemic rats. While plants
like Nauclea latifolia (Gidado et al., 2005), Rhinacanthus
nasutus (Rao and Naidu, 2010) and Veranonia
amygdalina (Michael et al., 2010) show hypoglycemic
effects in alloxin and streptozocin induced diabetic rats
but have no pronounced effects on blood glucose in
normoglycemic rats.
Conclusion
The ethanolic extract of the leaves of A. aspera validates
its use as a hypoglycemic drug. It can be used safely for
the treatment of diabetes type two, however in certain
cases, use of the drug by non-diabetic patient in high
doses will lower the normal glucose level. It is therefore
recommended that the plant should not be used in large
doses for the treatment of non-diabetic conditions.
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- ReferencesReferences24
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- [Show abstract] [Hide abstract] ABSTRACT: The effect of the leaf extract of Rhinacanthus nasutus in normal and Streptozotocin (STZ) induced diabetic rats was studied. Blood glucose levels were determined after oral administration of graded doses of R.nasutus (200 mg/kg) in fasted normal and diabetic groups. In both groups, 200 mg/kg of the extract, significantly reduced blood glucose levels 8 h after administration, which was consistent and time-dependent. The higher doses of 100 and 300 mg/kg did not affect significantly the blood glucose levels as preliminary experiments conducted earlier. The plant extract (methanolic) is given to the diabetic rats as well as normal rats orally by gavages. The blood glucose levels were increased when diabetes is induced in all the experimental rats. The methanolic extract of R. nasutus action was moderate in reducing the higher blood glucose levels in diabetic when compare with the normal rats. The normal rats' blood glucose levels did not change much when treated with plant extract alone. The plant extract was capable of ameliorating at the dosage of 200 mg/kg, hyperglycaemia in streptozotocin-induced diabetic rats and could be a potential source for isolation of new orally active agent(s) for anti-diabetic therapy.
- [Show abstract] [Hide abstract] ABSTRACT: The pharmacognostic profile of crude drug has key role in standardization for quality, purity and drug identification. Various pharmacognostic studies of Viola betonicifolia were carried out in order to establish parameters for its identification and to check adulteration or contamination by other species of Viola. Morphological characteristic of the whole plant and its anatomic parts were studied by organoleptic evaluation. Transverse sections of the root and petiole were prepared and studied. Powder drug study of the whole plant was also carried out and various structures of the powder drug were observed. This study also includes the numerical and quantitative leaf microscopy, ash values and extraction values. The pharmacognostic profile of V. betonicifolia was first time studied in this research paper.
- [Show abstract] [Hide abstract] ABSTRACT: This study investigated the antidiabetic activity of the various combinations (ratios) of metformin (50 mg/kg) and aqueous extracts of the leaves of Vernonia amygdalina (100 mg/kg). The ratios of Extract to Metformin were 1:1, 2 1, and 2:1 and distilled water (control, p.o.) were given to both normoglycemic and alloxan-induced diabetic Wister albino rats. Blood was withdrawn and tested at 0, 1, 3 and 6 hours. Results showed that the combinations of the extract and metformin caused more reduction in glycemia compared to any of the agents acting alone in either of the two categories of animals. The ratio of 1:2 caused the most significant (p%0.05) reduction in blood sugar (-66.07%) compared to distilled water (-7.2%). However, the ratio of metformin: extract (2:1) caused a reduction of -62.66% but was found a better combination considering the safety of the drugs. The combination of Vernonia amygdalina with metformin for the management of diabetes should be highly encouraged with a reduction in the dose of metformin and an increase in the dose of the plant extract to guarantee efficacy and safety.
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