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Diré et al.
Research Article
WORLD JOURNAL OF PHARMACEUTICAL
World Journal of Pharmaceutical and Medical Research
ISSN 2455-3301
AND MEDICAL RESEARCH
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WJPMR
ANALYSIS OF MEASUREMENT OF WEIGHT OF ANIMALS TREATED WITH A
FREEZE-DRIED AQUEOUS EXTRACT OF COSTUS SPICATUS.
Nascimento, C. C. H. C.1,2, Vasconcelos, S. D. D. De 1,3, Nascimento, S. F.1,2, Oliveira, J. F. F.2, Nogueira, R. I.4,
Barreto, A. S.1, *Diré, G. F.1,2
1
Laboratory of Chemical and Biological Analysis (LAQB), Foundation State University Center of the West Zone
(UEZO), Avenue Manuel Caldeira De Alvarenga, 1203, Campo Grande, Rio De Janeiro, 23070-200, Brazil.
2
Estácio De Sá University (UNESA), Rio De Janeiro, Brazil.
3
Federal Center of Technological Education Celso Suckow Da Fonseca - CEFET-NI. Rio De Janeiro, RJ – Brazil.
4
Empresa Brasileira De Pesquisa Agropecuária (Embrapa), Rio De Janeiro, Brazil.
*Corresponding Author: Dr. Diré, G. F.
Laboratory of Chemical and Biological Analysis (LAQB), Foundation State University Center of the West Zone (UEZO), Avenue Manuel
Caldeira De Alvarenga, 1203, Campo Grande, Rio De Janeiro, 23070-200, Brazil.
Article Received on 08/06/2016
Article Revised on 29/06/2016
Article Accepted on 19/07/2016
ABSTRACT
The Costus spicatus commonly called Brejo Cana in Brazil, is a medicinal plant found in coastal rainforests. The
rhizome of this plant is used for the treatment of complaints of the bladder and urethra and to expel kidney stones.
The infusion of the aerial parts is used to treat colds, sore throat, dysentery and diarrhea. Its use in traditional
medicine includes the use of leaves, stems and rhizomes as a diuretic and tonic. The antioxidant action of species
Costus is not yet well understood. However, studies show that many species of Zinziberaceae can sequester free
radicals. In some Western Hemisphere countries, especially in Brazil, Costus Spicatus is used in the form of
infusion for the treatment of kidney diseases and in order to reduce levels of blood glucose. The aim of this study
was to evaluate the effect of a freeze-dried aqueous extract of Costus spicatus in measuring the animal weight
(F1 mice) treated with said natural extract. The aqueous extract of Costus spicatus was administered daily orally (1
mL) with the aid of a gavage needle F1 male mice for 16 days. The data suggest that the aqueous extract studied
of Costus spicatus exhibit a weight maintenance effect of the treated animals. This effect may be related to action
mechanisms of certain phytomolecules which exhibit antioxidant and hypoglycemic action. There was no
reduced weight gain and food consumption in the treated group during the stages of the experiment.
KEYWORDS: Costus spicatus, antioxidant, hypoglycemic, plant extract, medicinal plant.
INTRODUCTION
Brazil is custodian of huge rich flora in raw material that
has aroused the interest of researchers from various
universities and institutions dedicated to herbal medicine.
The Amazon region is considered inexhaustible source of
therapeutic potential with the use of medicinal plants
(Martins, 2006).
The World Health Organization (WHO) has expressed its
position on the need to value the use of medicinal plants
in the health field, taking into account that 80% of the
population uses plants or their preparations as regards the
primary health care. Beside this, there is the participation
of countries development in this process, as have 67% of
the plant species in the world. In this context, only in
2011, phytotherapy in Brazil generated revenues R $ 1.1
billion. Therefore, the popular and institutional
interesthas grown to strengthen Phytotherapy in the
Unified Health System (SUS) as the use of medicinal
plants and their rituals provides an economical way
healing for most of the population, contributes
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significantly to the attention primary health (BRASIL,
2013a).
Since 2007, the SUS provides herbal plant derived, such
that currently offers the use of 12 herbal medicines of the
National List of Essential Medicines (RENAME)
available SUS (Holy Bramble, Guaco, Artichoke,
Aroeira, Cascara, Devil's claw, Isoflavone-of-soy, Cat
nail, Mint, Aloe, Willow, Plantago) (BRASIL, 2013b).
The Ministry of Health issued in February 2009, the
National Relationship Medicinal Plants of Interest to the
SUS (RENISUS). This list includes plants medicines that
have the potential to generate products of interest to
SUS. The list view was and is directing studies and
research that could subsidize the maintenance of herbal
ratio available for use by the population, with safety and
efficacy for the treatment of some diseases (BRASIL,
2009).
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Diré et al.
Ethnopharmacology meets these objectives, since its
basic purpose in phytochemical and pharmacological
studies to reconcile the information acquired from the
local communities that make use of medicinal flora in
order to provide data for research of affordable
medicines to world population, especially in developing
countries (Souza et al., 2012).
Ethnobotanical studies are important mainly in Brazil,
since its territory is home to one of the richest floras of
the globe, of which 99.6% are chemically unknown and
also one of the largest cultural diversities (Gottlieb et al.,
1996; Revilla, 2002).
The use of medicinal herbs in the treatment of various
diseases occurs for thousands of years. Ancient
civilizations already knew the medicinal power of some
plants and cultivated, passing the knowledge to every
generation. Over the years and the advent of medicine,
this knowledge has become devalued by health
professionals, who have begun to focus on allopathic
treatment. But today, science and health policies are
seeking to restore the use of medicinal plants by the
population (Feijó, et al., 2012).
Medicinal plants encompass a rich source of active
compounds that can neutralize snake venoms or
toxins. The Costus family has a broad popular use for
different
parts
of
the
American
continent.
Costus speciosus is an important medicinal plant widely
used in several indigenous medicinal formulations.
Costus spicatus is used by the Amazonian population to
treat inflammation, pain and other pathological
manifestations.
Costus pulverulentus
C.
Presl
(Costaceae), a species endemic to Mexico, is used for the
empirical treatment of cancer, pain, and inflammation
(Selim and Al Jaouni, 2015a; Picanço et al., 2016;
Alonso-Castro et al., 2016).
The ancestral use of herbs, over the years showed that
some plants have potentially dangerous substances. From
the scientific point of view, some research has shown
that many of these plants have aggressive substances and
therefore should be used with care, respecting their
toxicological risks (Veiga Jr. et al., 2005).
Despite all the advances, we see that the world market
for herbal medicines (understood as plant extracts, or a
mixture of various compounds, chemically and
pharmacologically quantified) grows gradually, and most
notable and unfortunate still is growing too more in
developed countries than in those who are not. The
improvement of pharmaceutical technology in
phytotherapy area allowed a better quality control of
drugs based on modern technology identification,
determination
and quantification
of chemical
compounds, making it possible to manufacture pesticides
safe, effective and fully reproducible effect. On the other
hand, advances in herbal research in pharmacological
level, toxicology and molecular have recognized that
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World Journal of Pharmaceutical and Medical Research
they have a full mechanism of action or partially cleared
up, with safe toxicological evaluation, and studies of preclinical pharmacology and clinical pharmacology carried
out according to the rules governing pure drugs
validation processes (Yunes et al., 2001).
In Brazilian folk medicine, the Costus spicatus tea is
used with depurative purposes, astringent and diuretic.
Boorhem (1999) describe the decoction of the vegetative
plant parts of the species active in the treatment of
vaginal irritation, leukorrhea and ulcers. Fresh juice
diluted stem is effective in the treatment of gonorrhea,
syphilis, nephritis, insect bites, bladder problems, and
diabetes. The sheets can be combined with Bonamia
ferruginea "vine-Tuira" (Choisy) Hallier in the form of
combinations (potions) in the treatment of malaria,
hepatitis and diabetes (Borras, 2003; Silva, 2004).
The objective of this study is to investigate the effect of
an aqueous extract of Costus spicatus in measuring the
F1 mice weight treated with the referred plant extract.
MATERIAL AND METHODS
Sample Plant and Collection
We used the leaves of Costus spicatus collected during
flowering (December-January) on the Sítio Nosso Sonho
(Site Our Dream, 22°54'48.0"S 43°54'02.6"W), the
municipality of Itacuruça, Rio de Janeiro, Brazil. The
choice of the time of collection was based on data from
the literature indicate that concentrations of flavonoids
substances tend to increase during this time. This is due
to the action of flavonoids as attractors of pollinators and
as co-pigments of anthocyanidins (Dourado & Ladeira
2008). The voucher specimen of the species will be
deposited in the Herbarium of the UFRJ National
Museum for botanical certification by an expert.
Preparation of extract
The leaves Costus Spicatus were dried in an oven with
circulating air at 40 °C, manually crushed and stored in
amber glass jar. The crude aqueous extract was obtained
by infusion of bark powder using distilled water as the
liquid extractant. The extract was lyophilized and stored
in amber glass bottle under refrigeration at a minimum
temperature of - 20 °C.
In experiments were used F1 mice, with about two
months old, male, kept under care related to food (food
and water ad libitum) except during the experimental
phase in temperature (22 ± 25 °C) and cycles 12 hours
light/ dark the Vivarium Foundation State University
Center of the West Zone (UEZO / Rio de Janeiro /
Brazil).
Procedures performed with animals were approved from
the previous analysis of a research project which was
submitted for review to the Ethics Committee for Animal
Use of the Foundation State University Center of the
West Zone (CEUA / UEZO).
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Administration Extract
The aqueous extract of Costus spicatus was administered
daily orally (1 mL) with the aid of a gavage needle F1
male mice for 16 days.
In the control group throughout the experiment, the
animals received food and water ad libtum, receiving no
oral administration of the plant extract.
The treated group consists of five animals receiving diet
ad libtum treated by oral administration of 1 mL of
lyophilized aqueous extract of Costus spicatus (10
mg/mL) for 16 days.
F1 mice Weight Analysis
The animals of the control and treated groups were
weighed on days 02, 10, 18 and 26 May, properly
stabilized in an balance (Semi-analytical balance class II
Bel Mark 220 - 220g), where measurements of their
weights were made.
The statistical analyzes were fragrant from the
processing of data of the statistical program Graph Pad
stat.
RESULTS
Table 1: Measuring the weights (g) of the control group animals.
Weight (g) control group F1 mice
Identification Days 05/02
05/10
1
36.3
35.7
2
41.4
41.7
3
35.4
36.8
4
34.8
37.0
5
39.1
40.2
05/18
37.3
42.5
34.2
36.5
40.8
05/26
36.7
41.4
34.2
37.3
39.4
.
Weight change (g) fasting of control animals (F1 mice) not treated with the extract of Costus spicatus.
Table 2: Measuring the weight (g) of animals treated.
Identification
1
2
3
4
5
Weight (g) mice treated group
Days 05/02
05/10
05/18
40.1
40.3
39.6
38.2
37.6
38.3
39.4
38.6
37.9
34.6
35.4
35.8
32.8
34.2
34.6
05/26
40.1
36.9
39.7
34.9
34.0
Weight change (g) F1 mice treated group fasting treated with the extract of Costus spicatus.
From the statistical analysis (ANOVA) was possible to
observe a value for p equal to 0.9897 is therefore not
significant. There was no reduced weight gain and food
consumption in the treated group during the stages of the
experiment.
Graph 1- Comparative analysis of the average weight of the animals of the control and treated groups.
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Diré et al.
In repeated dose 16-day oral toxicity study, the
administration of the extracts at the concentration of 10
mg/mL revealed no-treatment toxicity. The weight of the
animals no exhibited statistically significant effects, none
was adverse. Our findings suggest that the aqueous
extract of Costus spicatus is well tolerated for both single
and chronic administration. The oral no-observedadverse-effect level (NOAEL) for the extract was
10 mg/mL administered daily orally (1 mL) with the aid
of a gavage needle F1 male mice for 16 days.
In the graph above it can be seen the comparative
analysis of the average weight of the animals of the
control [A (05/02), B (05/10), C (05/18) and D (05/26)]
and treated groups [E (05/02), F (05/10), G (05/18) and
H (05/26)].
DISCUSSION
The use of medicinal plants has been used extensively in
the treatment and prevention of diseases in the
population in recent years. For millennia indigenous
peoples around the world have used plants for medicinal
purposes, and these skills were passed from generation to
generation to spread with the emergence of modern
civilizations. Thus, the global scientific community has
been working to confirm the popular use of medicinal
plants, thus contributing to the safe use and new drug
discovery (Rezende et al, 2004).
The Costus spicatus Jacq. S. W. (Costaceae /
Zinziberaceae), commonly called Cana do Brejo in
Brazil, is a medicinal plant found in coastal rainforests.
The rhizome of this plant is used for the treatment of
complaints of the bladder and urethra and to expel
kidney stones. The infusion of the aerial parts is used to
treat colds, sore throat, dysentery and diarrhea. Its use in
traditional medicine includes the use of leaves, stems and
rhizomes as a diuretic and tonic (Lorenzi & Matos,
2008).
Diosgenin, a naturally occurring steroid saponin found
abundantly in Costus speciosus, and also found in Costus
spicatus, is a well-known precursor of various synthetic
steroidal drugs that are extensively used in the
pharmaceutical industry. Selim and Jaouni (2015b)
suggested that diosgenin isolated from C. speciosus
possess anticancer and apoptotic effects on cell
proliferation, and therefore, can be used as
pharmaceuticals drugs. In relation to the effects
described we can speculate that the extract under study
could be able to maintain its integrity thereby
contributing to the maintenance of cellular and tissue
homeostasis thus contributing to the preservation of
animal weight.
Picanço et al., (2016) reported that an extract of Costus
spicatus could present a central mechanism of inhibition
of pain, and can also inhibit prostaglandin synthesis.
These findings corroborate the traditional administration
of decoction of Costus spicatus to treat inflammatory
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World Journal of Pharmaceutical and Medical Research
diseases, including those caused by Bothrops atrox
poisoning. In our experiment, we found that an aqueous
extract of Costus spicatus was not able to induce changes
in animal weights that were treated with the natural
extract referred.
Perera et al., (2016), reported that the methanolic extracts
Costus Spicatus sheets showed inhibitory activities in
vitro α-glucosidase, fructosamine formation, glycation
and glycation induced cross-linking of proteins. These
results provide scientific evidence to support the use of
Costo spicatus sheets for hypoglycemic effects with an
additional advantage in delaying the glycation of
proteins. These findings related to the hypoglycaemic
effect of Costus species could justify maintaining the
weight of the treated animals in our study with the
natural extract, as phytochemicals components present in
the constitution of the aqueous extract of Costus spicatus
could help in preserving the structural tissue proteins, as
well as glycogen in muscle and liver.
Nascimento et al., (2016a) reported that phytochemical
compounds of Costus spicatus present in the leaves of
plant include glycosides, tannins, saponins, terpenoids,
phenolics, flavonoids, alkaloids as well as eremanthin
could optimize sugar uptake in the liver. These feats
could induce insulin secretion and release from cells, as
well as stimulates the tissue's insulin sensitivity leading
to an increase of the tissues' glucose uptake, storage, and
oxidation.
In a study Hardikar et al., (2016) described that active
insulin-like protein (ILP) from Costus igneus has a
potent hypoglycemic property in STZ-induced diabetic
Swiss mice. Their study revealed that ILP acts via insulin
signaling pathway and can be used as oral insulin
mimetic.
Nascimento et al (2016b) reported that an aqueous
extract of Costus spicatus studied has a potential
hypoglycemic action which could be related to the
synergic action of molecules with antioxidant profile
which can enhance the secretion of insulin by the
pancreas and the increase in absorption of tissue glucose
level. Based on evidence from the analysis of the results,
they suggested that the effect of decreasing glucose
Costus Spicatus extract could be associated with the
enhancement of insulin release from pancreatic islets and
enhancing peripheral glucose utilization. This fact could
support our findings due to the with the preservation of
weight of animals for hypoglycemic and antioxidant
effects related to Costus spicatus extract.
Oxidative stress is developed due to susceptibility of
biological substrates to oxidation by generation of free
radicals. In degenerative diseases, oxidative stress level
can be reduced by antioxidants which neutralize free
radicals. Pai Kotebagilu et al., (2014) observed that
extract of Costus showed highest inhibition of oxidation
and correlated well with the polyphenol and flavonoid
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content. They concluded that antioxidants from
medicinal plants are capable of inhibiting oxidation in
biological systems, suggesting scope for their use as
nutraceuticals.
In another study Nascimento et al., (2015) speculated
that an aqueous extract of Costus spicatus features
phenolic compounds with anti-hemolytic action and
saponins molecules which would be possibly related with
the hemolytic effects related with the natural extract
referred studied. These findings may reinforce the
understanding that certain phytochemicals components
present in Costus spicatus extract could help in the
preservation of tissue protein structures which are
important for the morphological and physiological
maintenance of animal tissues collaborating in this way
to maintain the weight of the treated animals.
CONCLUSION
The data suggest that the aqueous extract studied Costus
spicatus exhibit a weight management effect of the
treated animals. This effect may be associated with
certain mechanisms phytomolecules which exhibit
antioxidant and hypoglycemic action. From the chronic
treatment performed with an aqueous extract of Costus
spicatus we can speculate that the referred natural extract
has no adverse effects and no poses the health risk in
relation to oral toxicity study.
World Journal of Pharmaceutical and Medical Research
8.
9.
10.
11.
12.
13.
ACKNOWLEDGEMENT
We thank the Foundation Carlos Chagas Filho Research
of the State of Rio de Janeiro (FAPERJ) for supporting
this research.
14.
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