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Morphoanatomy and pharmacognostic study of the wood of Croton echioides, the Northeastern Marapuama

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  • Universidade Anhanguera de São Paulo - Unian-SP

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Croton echioides Baill., Euphorbiaceae, is a small tree found in Bahia, Northeastern Brazil. Its stem bark has been widely sold as an aphrodisiac and tonic, as a substitute for the roots of Ptychopetalum olacoides Benth. Olacaceae, the Amazon Muira Puama or Marapuama, and C. echioides is characterized as the "Northeastern Marapuama". This contribution describes a morphoanatomical analysis and pharmacognostic study of stem bark of this species. The stem has a thick cortex with compound starch grains and laticifers; a sclerenchymatic sheath which consists of brachysclereids with large crystals externally to the phloem, and abundant fiber in the secondary xylem, as the main features of the species. The data obtained for water content (9.26±0.07%), water-soluble extractives (3.92±0.19%), total ash (1.24±0.06%) and acid-insoluble ash (0.16±0.01%), together with the chromatographic profile obtained by TLC, contribute to the quality control and standardization for the plant drug. The pharmacological screening indicated LD50 values above 500 mg/kg orally and equal to 500 mg/kg by the i.p. route, as well as some stimulant potential, depending on the dose.
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946
ISSN 0102-695X
http://dx.doi.org/10.1590/S0102-
695X2012005000062
Received 23 Jan 2012
Accepted 11 Apr 2012
Available online 15 May 2012
Revista Brasileira de Farmacognosia
Brazilian Journal of Pharmacognosy
22(5): 946-956, Sep./Oct. 2012 Morphoanatomy and pharmacognostic
study of the wood of Croton echioides, the
Northeastern Marapuama
Cláudio R. Novello,1 Luís C. Marques,2 Cristine R. Miyazaki,2
Maria A. Milaneze-Gutierre,3 Daniela S. Carneiro-Torres,4
Maria H. Sarragiotto,5 João C. P. de Mello*,1
Abstract: Croton echioides Baill., Euphorbiaceae, is a small tree found in Bahia,
Northeastern Brazil. Its stem bark has been widely sold as an aphrodisiac and
tonic, as a substitute for the roots of Ptychopetalum olacoides Benth. Olacaceae,
the Amazon Muira Puama or Marapuama, and C. echioides is characterized as the
"Northeastern Marapuama". This contribution describes a morphoanatomical analysis
and pharmacognostic study of stem bark of this species. The stem has a thick cortex
with compound starch grains and laticifers; a sclerenchymatic sheath which consists
of brachysclereids with large crystals externally to the phloem, and abundant  ber in
the secondary xylem, as the main features of the species. The data obtained for water
content (9.26±0.07%), water-soluble extractives (3.92±0.19%), total ash (1.24±0.06%)
and acid-insoluble ash (0.16±0.01%), together with the chromatographic pro le
obtained by TLC, contribute to the quality control and standardization for the plant
drug. The pharmacological screening indicated LD50 values above 500 mg/kg orally
and equal to 500 mg/kg by the i.p. route, as well as some stimulant potential, depending
on the dose.
Keywords:
Croton echioides
Euphorbiaceae
morphoanatomy
Northeastern Marapuama
pharmacognostic quality control
Introduction
Products of natural origin with reputed
aphrodisiac effects have long been of great interest for
therapeutic uses and also to international pharmaceutical
companies, and have been sought after and studied
for millennia. Some of the sought-after species have
proved to be effective, as occurred with yohimbe bark
(Corynanthe johimbe K. Schum., Rubiaceae), the roots of
Korean ginseng (Panax ginseng C.A. Mey., Araliaceae),
and cantharidin obtained from insect cantharides (Lytta
vesicatoria L., Meloidae), among others (Riley, 1994;
Sandroni, 2001).
A Brazilian plant with popular interest and
commercial potential for its aphrodisiac effects is Muira
Puama or Marapuama, the root bark of Ptychopetalum
olacoides Benth., Olacaceae. This plant drug has been
marketed and exported since the beginning of the
twentieth century, when it was sometimes counterfeited
by substitution of guava roots (Silva, 1925). This was
later formalized in the rst edition of the Brazilian
Pharmacopoeia (Silva, 1926).
Several studies have provided evidence of the
effects of this plant drug on the central nervous system.
Initially it was found to show effects as an anxiogenic
(Silva et al., 2002), anticholinesterase (Siqueira et al.,
2003), neuroprotective (Siqueira et al., 2004), facilitation
of memory recovery in young and old mice (Silva et al.,
2004), antioxidant (Siqueira et al., 2007), antidepressant
(Paiva et al., 1998; Piato et al., 2009), and anti-stress
(Piato et al., 2010). Its aphrodisiac potential has not been
con rmed experimentally or clinically, although some
of its effects, such as antidepressant and antioxidant
activity, may improve sexual performance (Feldman et
al., 1994).
Interest in these effects and the folk wisdom of
the Brazilian Amazon have kept this species in evidence,
together with catuaba and other plants (Charam, 1987;
Pozzoli, 1999). Consequently, it has continued to be
collected in the wild, marketed, and exported for decades.
The continual exploitation of this Amazon endemic
species, the use of roots, and the lack of incentives
to cultivate it to supply this demand must have led to
signi cant reduction of its natural populations.
Article
1Departamento de Farmacia, Universidade Estadual de Maringá, Brazil,
2Universidade Bandeirante de São Paulo, Brazil,
3Departamento de Biologia, Universidade Estadual de Maringá, Brazil,
4Universidade Estadual de Feira de Santana, Brazil,
5Departamento de Química, Universidade Estadual de Maringá, Brazil.
Morphoanatomy and pharmacognostic study of the wood of Croton
echioides, the Northeastern Marapuama
Cláudio R. Novello et al.
Rev. Bras. Farmacogn. Braz. J. Pharmacogn. 22(5): Sep./Oct. 2012 947
When such a situation arises, usually the market
seeks alternative sources of supply, eventually triggering
the replacement of the true species by another with the
same indications, although it may not always be chemically
similar, as occurred with the catuaba (Marques, 1998a). In
the case of P. olacoides, the commercial substitution led
to exploitation of stems, which were initially offered as
being from the species itself, as a strategy to prevent the
destruction of the specimens by removal of the roots, as
reported by plant drug distribution companies. However,
it was gradually found that these stems came from the
interior of the state of Bahia, probably originating
from some species of Croton, Euphorbiaceae, and were
completely different from the Amazon Marapuama
(Tobias et al., 2007).
The large and diverse genus Croton L. occurs
in tropical and subtropical regions worldwide (Webster,
1993), and according to Govaerts et al. (2000) contains
about 1200 species. The main compounds found
in the genus Croton are clerodane-type diterpenes,
avonoids, and indole alkaloids (Matos, 2011). Many
clerodane diterpenes show biological activity, including
antibiotic, antitumor, insecticidal, anti-inammatory,
antinociceptive, hypoglycemic, antiulcer (Maciel et al.,
1998), and cardiovascular (Silva et al., 2005).
P. olacoides was reported initially to contain
behenic acid ester and β-sitosterol (Auterhoff & Pankow,
1968), free fatty acids, and no alkaloids (Toyota et al.,
1979), as well as several steroids, the triterpene lupeol,
avonoids, and saponins (Maul et al., 1998). More
recently, Colombo et al. (2010) found vanillic acid,
protocatechuic acid, and theobromine in the stems of P.
olacoides. However, the presence of essential oils, tannins
and alkaloids was reported by Bucek et al. (1987), as
well as several clerodane diterpenes by Tang et al. (2009,
2011).
By linking these contradictory phytochemical
observations with the reported problem of adulteration
of commercial species in the Brazilian market, which
also extrapolates to the international market, even as
raw material for scientic research, it is apparent that
some of these studies in fact assessed the stems of the
"Northeastern Marapuama".
Taking into account the lack of strict control of the
harvesting, identication, and botanical characterization,
for chemical or pharmacological quality control for this
species of Croton, this study aimed to collect and identify
unambiguously the species used as the "Northeastern
Marapuama", C. echioides Baill. In addition, we sought to
describe the morphoanatomy of the plant drug (stems) and
conduct a pharmacognostic analysis, establishing the rst
scientic specications for this raw material.
Materials and Methods
Plant material
Through contacts with suppliers of plant drugs
for the Brazilian market, we tracked the source location
of stems offered as P. olacoides in commercial batches
of raw materials. This effort led to José Maria Fernandes
Monteiro from the municipality of Lagoa Real, state of
Bahia, and we visited him personally to collect, plant
material, in December 2008. Mr. Monteiro is a leading
supplier of plant drugs for the Brazilian pharmaceutical
market.
The owering aerial parts of Croton echioides
Baill., Euphorbiaceae, were collected at Lagoa
Real under collection authorization No. 11995-3 of
02.11.2010, registered as IBAMA No. 1844493, under
the responsibility of João Carlos Palazzo de Mello. We
also collected samples of stems (plant drugs) for use in
further studies.
Botanical identication and herborization
The botanical material was mounted on
exsiccates, which were deposited in the herbarium of
the Universidade Estadual de Feira de Santana, under
HUEFS number 139049, and in the herbarium of the
Universidade Estadual de Maringá, under HUEM number
19433. The species was identied by Prof. Dr. Daniela
Santos Carneiro Torres, with the aid of the key by Silva
et al. (2009).
Pharmacobotany characterization
The organoleptic characterizations, and
macroscopic and microscopic examinations of stem
segments were performed according to standard
methodology (Oliveira et al., 2003; Farmacopeia
Brasileira, 2010). Pieces of commercial stems were
evaluated, described, and photographed. Young (up to 1
cm diameter) and mature (over 1 cm in diameter) stem
segments were dried and evaluated with the naked eye,
with regard to the macroscopic structure, rhytidome
color, and internal tissues. For the anatomical analysis,
fresh stems were cut into smaller portions, xed in FAA
50, and conserved in 70% ethanol (Johansen, 1940).
The histological sections were made on several planes,
freehand with the aid of stainless-steel blades, and then
bleached with sodium hypochlorite (30% commercial)
for approximately 5 min, stained with safranin and Astra
blue (1% aqueous solutions in the ratio 9:1 v/v) (Kraus
& Arduin, 1997), and mounted as semi-permanent slides
with glycerin jelly. For the preparation of the illustrations,
the tissues were photographed with a digital camera
attached to an Olympus microscope, using the software
Image-Pro Plus 4.5.
Morphoanatomy and pharmacognostic study of the wood of Croton
echioides, the Northeastern Marapuama
Cláudio R. Novello et al.
Rev. Bras. Farmacogn. Braz. J. Pharmacogn. 22(5): Sep./Oct. 2012
948
Pharmacognostic quality control assays
Samples of stems of C. echioides (about 10 kg)
were cut into small pieces, dried at room temperature in
the PALAFITO-UEM Laboratory, ground in a hammer
mill (Tigre ASN5), and stored hermetically sealed and
protected from heat and light. The granulometry of the
powder was measured by sieves of mesh sizes 150, 180,
212, 300, 600, and 850 μm (Bertel AGT-01). The levels
of total ash, acid-insoluble ash, and loss on drying were
determined according to methods described by Brazilian
Pharmacopeia (Farmacopeia Brasileira, 2010) as were
the water-soluble extractives (WHO, 1998), all from the
samples of powdered stems.
For thin layer chromatography (TLC), ground
or powdered plant drug (1 g) was subjected to reux
with 100 mL of methanol, ltered and concentrated to 10
mL. This solution (20 μL) was applied to silica gel F254
plates (Merck), eluted with n-hexane: dichloromethane:
methanol (6:3:1, v/v), and revealed with sulfuric
anisaldehyde upon heating to 110 °C, and the retention
factors (Rf) were determined.
Pharmacological screening of the crude extract
We used male Swiss mice aged 2-3 months and
weighing from 30-40 g, from the Center for Development
of Experimental Models of the Universidade Federal de
São Paulo. The animals were kept for a week before
the experimental procedures at the Laboratory of
Experimental Pharmacology of Uniban, in groups of
ten animals per box (24±3 °C) with a controlled light/
dark cycle (12 h) and fresh water and food ad libitum,
up to 120 min before the start of the experiments. All
experimental procedures were performed according to
the ethical principles in animal experimentation adopted
by Uniban protocol (No. 20090161). The plant extract
was dissolved in water with drops of Tween 80 at the
time of testing.
The extracts were prepared from powdered
stems of the plant (7.6 kg) by Ultra-Turrax (Ika Works,
UTC115KT) using 70% (v/v) hydroalcoholic solvent
in the proportion of 10% (w/v). The extraction was
performed for 5 min of stirring followed by 2 h of rest,
successively, within a period of 48 h. The solvent was
removed in a rotavapor under reduced pressure, frozen,
and lyophilized to provide the crude extract (CE).
Groups of ve young male mice received various
doses of CE (1, 10, 100, and 500 mg/kg) by i.p., and 500
mg/kg of CE orally (gavage), together with a control
group. Immediately thereafter the mice were placed in
wire cages (ve per cage), and were observed at the times
of 5, 15, and 30 min, 1, 2, 4, and 24 h, noting all behaviors
pertinent to the expression of some pharmacological and/
or toxic effect. The animals were weighed before the
experiment and again after fourteen days of observation.
The animals were evaluated according to the protocol
established by Carlini (1972): urination, defecation,
abdominal constrictions, rough hair coat, motor activity,
tremors, convulsions, ataxia, loss of posture reex, pain
sensitivity, ptosis, Straub tail, abduction of the hind legs,
signs of stereotypy, yawning, sleep, excessive grooming,
muscle tone, lacrimation, exophthalmia, salivation, and
death.
Results and discussion
Botanical identication
Review of the literature showed that, until the
present, C. echioides had not been properly identied and
related to the commercial material available throughout
the country, and also as raw material for export, since
invariably it was labeled and offered as Ptychopetalum
olacoides. Thus, the systematic examination proved
crucial to unequivocally identify the plant material
offered in Brazil as ‘Marapuama’.
These observations have claried the
identications of the main species known as Brazilian
aphrodisiacs, including Catuaba stem bark of Trichilia
catigua A. Juss. (Marques, 1998a); Brazilian Ginseng
roots Pfafa glomerata (Spreng.) Pedersen (Marques,
1998b); and now Croton echioides as the "Northeastern
Marapuama", available commercially as a substitute for
the true "Marapuama", P. olacoides.
Croton echioides is popularly known in the
northeast region of Brazil as "quebra-faca", "caatinga-
branca" and "canela-de-velho". It is restricted to the
Caatinga biome, occurring in the states of Piauí, Ceará,
Rio Grande do Norte, Pernambuco, Paraíba, Alagoas,
Bahia, and Minas Gerais in arboreal to shrubby caatinga
vegetation, in sandy and clay soils, with rocky outcrops
(Figure 1). As do all species of Euphorbiaceae, C. echioides
has unisexual owers and schizocarp fruit, tricarpellar
with only one ovule per locule. It is differentiated from
other species of Croton of the Caatinga biome in having
leaves with the margin entire, the presence of two short-
stipitate glands between the petiole and leaf blade,
staminate owers with spatulate petals, and pistillate
owers that are sessile and sparsely arranged along the
inorescence.
Organoleptic, macroscopic and microscopic
characterization
The samples are marketed in stem fragments
of various sizes, 10-15 cm long and about 1-4 cm wide.
The rhytidome is mixed in color, ranging from light
gray to light brown with dark-brown spots; there are a
few thick cracks, with no regular pattern, but generally
Morphoanatomy and pharmacognostic study of the wood of Croton
echioides, the Northeastern Marapuama
Cláudio R. Novello et al.
Rev. Bras. Farmacogn. Braz. J. Pharmacogn. 22(5): Sep./Oct. 2012 949
longitudinal. In median longitudinal section, the cork
appears thin, strongly adhered to other tissues. The wood
is light brown, with the central, medullar, dark brown
along its entire length; in transverse view, the medullary
tissue shows as a central dark spot (Figure 2 and 3).
These characteristics are consistent with the strains of C.
echioides collected in the municipality of Lagoa Real, as
are the organoleptic characteristics, in which the bark has
its own aromatic odor and slightly bitter taste; the wood
is odorless and tasteless.
Figure 1. Map of geographical distribution of Croton echioides
Baill. Gray delineates the Caatinga biome.
Figure 2. Stem segments of Croton echioides.
The optical-microscopy analysis revealed that
the stem of C. echioides shows a periderm composed of
6-7 strata of quadrangular cells in cross section (Figures
4A and 5A), somewhat elongated longitudinally (Figures
4B and 5B) and 10 to 12 strata can occur at sites of
defoliation. The cells of the periderm are thicker in the
periclinal walls (Figure 5C) than in the opposite walls, and
have different dimensions when viewed in longitudinal
tangential section (Figure 5D).
The stem cortex of C. echioides is thick, with
approximately twenty strata whose cells vary in diameter
(Figures 4A and 5A). A longitudinal section revealed
somewhat longitudinally elongated cortical cells,
sometimes isodiametric, but also with no uniformity
in their dimensions (Figures 4B, 5B, and 6B). This
tissue contains compound starch grain (Figure 6C) and
abundant laticifers (Figures 4A, 4B, 5A, 5B, and 6A, 6C,
and 6D). According to Farias et al. (2009), the presence
of latex in the cortical portion of the stem is common in
many species of the genus Croton. Although few studies
describe the anatomical tissues of the stem of species of
Brazilian Croton, Silva (2006) reported the occurrence of
laticifers located near the region of the stem phloem of C.
oribundus. When comparing the Marapuama samples
available on the market, the presence of laticifers, which
is common in Euphorbiaceae, in contrast to Olacaceae
species, serves as a distinctive character between
P. olacoides and C. echioides, although it does not
distinguish the latter from other members of the genus
Croton.
More internally, and involving the vascular
system, there is a thick sclerenchymatous sheath, which
consists of brachysclereids. In young stem, has 5-6
layers of this cells (Figure 6A) and 10-17 layers in older
stem (Figure 7A). The sclerenchymatic cells are more
homogeneous in shape when viewed in cross section
(Figures 7A, 7B, and 7C), but show different shapes,
tending to be elongated in longitudinal section. In its thick
walls, lamellations (Figure 7B) and branched punctations
are evident (Figure 7C), similar to those observed by
Silva (2006) in the secondary stem of C. oribundus
and C. macrobothrys, although in these species they
are arranged in a discontinuous ring, interspersed with
parenchyma cells. In addition, according to Thomas
et al. (1995) in Hevea brasiliensis these sclereids are
distributed diffusely or in bundles, from near the periderm
to the phloem region. Both the brachysclereids and the
cells that compose the parenchyma rays that permeate the
sclerenchymatous sheath contain large crystals, which
are polygonal to typical prismatic in shape (Figures 6A,
7A, and 7B). The presence of sclerenchyma with large
crystals in the secondary phloem was also reported by
Farias et al. (2009) in C. draco var. draco, a Mexican
species. The druses observed by Silva (2006) in the
cortical parenchyma of C. oribundus were not found in
C. echioides.
The secondary xylem of C. echioides rarely
contains parenchyma cells (Figures 8A, 8B and 8C),
which is not common for other members of this taxonomic
group. In the secondary xylem, growth rings were evident
(Figure 8A) as were vessel elements, most often solitary,
although multiple pairs and clusters may also occur
(Figures 8A, 8B, and 8C). In these vessel elements the
punctuations are areolate and the alternating type (Figure
8F). The bers that compose the secondary xylem of C.
echioides are abundant, with very thick walls, giving
them a small lumen (Figures 8B and 8C), and a high
degree of rigidity to the mature stems of this species.
Morphoanatomy and pharmacognostic study of the wood of Croton
echioides, the Northeastern Marapuama
Cláudio R. Novello et al.
Rev. Bras. Farmacogn. Braz. J. Pharmacogn. 22(5): Sep./Oct. 2012
950
The gelatinous bers, common in species of Croton, also appear in abundance in the secondary xylem of youngers
stems.
The parenchyma rays of secondary xylem are uniseriate and are almost obliterated in mature stem segments
(Figures 8C, 8D, and 8E). In this gures can also be seen crystals, as in the phloem, present in nearly all the cells of
the ray parenchyma.
Figure 3. Exsiccata (A) and stem segments of Croton echioides (B). The arrow indicates a typical transverse projection of the stem.
Figure 4. Sections of Croton echioides stem. General appearance in cross section (A) and tangential longitudinal section (B). Arrows
indicate laticifers. co: cortex, pe: periderm; ph: phloem; ss: sclerenchymatic sheath; sxy: secondary xylem.
Morphoanatomy and pharmacognostic study of the wood of Croton
echioides, the Northeastern Marapuama
Cláudio R. Novello et al.
Rev. Bras. Farmacogn. Braz. J. Pharmacogn. 22(5): Sep./Oct. 2012 951
Figure 5. Sections of the more peripheral stem tissues of Croton echioides. Details of the periderm and cortex in cross section
(A) and tangential longitudinal section (B). Details of the periderm in cross section (C) and longitudinal tangential section
(D). The single arrows indicate laticifers, and the double arrow indicates the phellogen. co: cortex; pe: periderm.
Figure 6. Sections in the cortical region of young stems of Croton echioides. Detail of the cortex in cross section (A), longitudinal
tangential section (B) radial longitudinal section (C). D shows the arrangement of laticifers in the cortex. Arrows indicate laticifers.
co: cortex; csg: compound starch grain; pc: prismatic crystal; ph: phloem; ss: sclerenchymatic sheath.
Morphoanatomy and pharmacognostic study of the wood of Croton
echioides, the Northeastern Marapuama
Cláudio R. Novello et al.
Rev. Bras. Farmacogn. Braz. J. Pharmacogn. 22(5): Sep./Oct. 2012
952
Figure 7. Sections in the sclerenchymatic sheath region of older stems of Croton echioides. General aspect (A) and details of
brachysclereids in cross section (B e C). Arrows indicate prismatic crystal. bp: branched punctations; co: cortex; csg: compound
starch grain; la: lamellations.
The pith, even in young stem, is compose by
sclerenchymatic sheath which consists of brachysclereids,
with a square to polygonal shape in cross section (Figures
9A and 9B), most of them slightly elongated compared to
the longer axis of the organ (Figures 9C and 9D). In this
cell type with very thick walls, the punctuations are, in
most cases, branched (Figure 9D).
Pharmacognostic quality-control data
The results for the analysis of the
physicochemical quality control of the stems of C.
echioides are seen in Table 1. The levels are within
the parameters found in most plant drugs in the
pharmacopeia, with the exception of the particle-size
test, which is specific for this plant.
Table 1. Quality-control analysis of powdered stem bark of
Croton echioides.
Assay Results
Powder granulometry 360±10 μm (n=3)
Water content 9.26±0.07% (n=5)
Water-soluble extractives 3.92±0.19% (n=5)
Total ash 1.24±0.06% (n=5)
Acid-insoluble ash 0.16±0.01% (n=5)
The thin-layer chromatographic analysis
showed a different chromatographic profile, as revealed
with sulfuric anisaldehyde (Figure 10). In addition,
there were light-brown and purple spots with the same
Rf values 0.24 and 0.52, compared to the reference
compounds diterpene [15,16-epoxy-3,13(16),14-neo-
clerodatrien-17-carboxy-18-methyl carboxylate] and
Morphoanatomy and pharmacognostic study of the wood of Croton
echioides, the Northeastern Marapuama
Cláudio R. Novello et al.
Rev. Bras. Farmacogn. Braz. J. Pharmacogn. 22(5): Sep./Oct. 2012 953
Figure 8. Sections of the secondary xylem of Croton echioides. General aspect (A) and details of the tissue organization (B and C)
in cross sections. Details of the xylem tissue in tangential longitudinal sections (D, E and F). ap: alternating punctations; fb: ber; pc:
prismatic crystal; pr: parenchyma ray; ve: vessel element.
lupeol, respectively, as shown in Figure 9. Notably,
lupeol is also found in P. olacoides (Auterhoff &
Momberger, 1971; Ito et al., 1995), and therefore is not
a useful standard for the differentiation of the species P.
olacoides and C. echioides.
Pharmacological screening results
The CE showed a preliminary stimulatory effect
at low doses, by oral and i.p.; toxicity was evident for the
larger i.p. doses, with 100% mortality after administration
of 500 mg/kg (Table 2). Given this prole, it is suggested
that more specic pharmacological tests be used with
doses ranging from 1 to 100 mg/kg, in order to determine
more precisely the levels of toxicity. One case of Straub
tail was observed, which indicated stimulation of opioid
receptors; it would be interesting to apply tests for
analgesia or others aimed at evaluating this potential
effect. Most of the groups lost weight, although not in
statistically signicant amounts.
Conclusion
The data presented here conrmed that the stems
of Croton echioides are the "Northeastern Marapuama"
that is widely marketed within and outside Brazil. The
pharmacognostic data obtained in this study resulted in
an effective basic characterization and standardization
of this plant material, contributing to the differentiation
from the plant drugs obtained from roots or stems of
Ptychopetalum olacoides.
Morphoanatomy and pharmacognostic study of the wood of Croton
echioides, the Northeastern Marapuama
Cláudio R. Novello et al.
Rev. Bras. Farmacogn. Braz. J. Pharmacogn. 22(5): Sep./Oct. 2012
954
The LD50 values above 500 mg/kg orally and
equal to 500 mg/kg by the i.p. route indicated that the
species has low acute toxicity (Zucker, 1985), although
new pharmacological and toxicological studies would be
appropriate to better understand these aspects. Therefore,
further investigations are needed to conrm whether
there are effective therapeutic and pharmacological
similarities between the two Brazilian species popularly
named "Marapuama".
Figure 9. Sections of the primary xylem and medulla from young stem segment of Croton echioides. General aspect (A) and
details of primary xylem (B) in cross sections. Details of the medullary tissue in tangential longitudinal sections (C and D).
bp: branched punctations; me: medulla, pxy: primary xylem poles; sxy: secondary xylem.
Table 2. Results of the pharmacological screening test of the CE of Croton echioides in young male mice (n=5 per group).
parameters oral route intraperitoneal route
control 500 mg/kg 1 mg/kg 10 mg/kg 100 mg/kg 500 mg/kg
mortality no no no no no 100%**
abdominal contortions no no no no yes (+) yes (++)
tremors no no no no yes (+) yes (++)
motor activity normal increase (+) increase (+) increase (++) decrease (-) death
climbing behavior no no yes yes no death
straub tail no no no no yes* death
initial weight (g) 40.4±0.8 45.2±4.5 39.3±3.0 42.3±3.0 40.1±3.0 death
nal weight (g) 39.2±1.0 37.6±5.0 34.2±12.0 42.3±2.0 39.3±2.0 death
Anova: not signicant; *one animal; **5 min.
Acknowledgments
This work was supported by the Brazilian
granting agencies CNPq, CAPES/Proap, INCT_if,
FINEP and Fundação Araucária. The authors are grateful
to Dr. Janet W. Reid, JWR Associates, Trumansburg,
New York, for the English language revision. In
memorian Prof. Dr. Gentil José Vidotti.
Morphoanatomy and pharmacognostic study of the wood of Croton
echioides, the Northeastern Marapuama
Cláudio R. Novello et al.
Rev. Bras. Farmacogn. Braz. J. Pharmacogn. 22(5): Sep./Oct. 2012 955
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*Correspondence
João Carlos Palazzo de Mello
Departamento de Farmácia. Universidade Estadual de Maringá
Avenida Colombo, 5790, BR-87020-900, Maringá-PR, Brazil
mello@uem.br
Tel. + 55 44 3011 4816
Fax: +55 44 3011 5050
... In addition, plants of this genus have been studied for their anti-cancer action. The extract of C. campestris had its antitumor activity confirmed, inhibiting the growth of lung tumor cells (Monteiro, 2012), and some indole alkaloids of C. echioides showed in vitro action to combat human colon carcinoma (Mello et al., 2010 andNovello et al., 2012). ...
... According to Martins (2018), the hexanic extract of Euphorbia tirucalli, also from the Euphorbiaceae family, as well as C. echioides, presented itself as promising for the development of new anticancer drugs. Possibly this activity is again associated with the antioxidant activity of the extract, which was confirmed by Novello et al. (2012). ...
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The search for compounds with anticancer effects is of paramount importance today due to the high incidence of the disease. The Euphorbiaceae family is known for having compounds with therapeutic properties, one of its genera being Croton. It has several species, which contain compounds already known for their biological activities, presenting anti-inflammatory, antimicrobial and anticancer properties. Thus, the cytotoxicity/antiproliferative activity of semi-purified fractions and compounds isolated from Croton echioides in liver tumor cells of Rattus norvegicus (HTC) was evaluated by the MTT test. The semi-purified fractions showed cytotoxicity at concentrations above 200 µg/mL, at 24, 48 and 72 hours, reaching cell viability of 24.78% [400 µg/mL] at 24 hours, 12.79% [500 µg/mL] at 48 hours and 10.57% [300 µg/mL] at 72 hours. For the isolated compounds, lupeol had a cytotoxic effect in all concentrations (1, 5, 10, 15, 20, 40, 60, 80 and 100 µg/mL) and tested times (24, 48 and 72 hours), reaching minimum viability of 4.37% [100 µg/mL], within 72 hours. The clerodan diterpenes CEH-1 and CEH-4 also showed antiproliferative activity, with minimum viability of 36.19% [100 µg/mL] over 72 hours and 21.33% [100 µg/mL] over 48 hours, respectively. However, the clerodan diterpenes CEH-2 and CEH-3 did not shows a cytotoxic effect for HTC cells. Thus, there is a cytotoxic/antiproliferative potential of C. echioides against tumor cells, with targeted to mitochondrial enzymes, associated with cell proliferation, indicating that this species deserves prominence in the search for new molecules for the treatment of cancer.
... by Luchi (2004), because the cellulosic lamellae ('G' layer) detach from the lignified layer. Gelatinous fibers are present in many genera of Euphorbiaceae, as reported by Mennega (2005), being common in Croton species, and abundant in the secondary xylem of young C. echioides Baillon stems (Novello et al. 2012). In this cortical portion, we also found amyloplasts (Fig. 4c) and idioblasts with druses ( Fig. 4d) that reacted positively with 60% chloral hydrate with 25% sulfuric acid. ...
... Energy-dispersive X-ray spectroscopy analysis of these crystals showed calcium (20.3%), carbon (27.4%), and oxygen (41.3%) peaks (Fig. 5). (Novello et al. 2012), but were abundant in C. lechleri and C. lanjouwensis (Amaral et al. 2005). Idioblasts with dense content reacted positively with ferric chloride and Sudan IV, confirming the presence of polyphenols (more abundant) and lipid substances, respectively, which are more frequent in thicker stem bark. ...
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Croton floribundus, Euphorbiaceae, popularly known as “capixingui” is a native tree of the Atlantic Forest. In folk medicine, the tea of C. floribundus stem bark is used for the treatment of leukemia, tumors, and syphilis. The aim of this work was to describe the morphological and anatomical characteristics of C. floribundus stem bark and to establish parameters for its quality control. Accordingly, different analyses were performed, including organoleptic, morphological, anatomical, and histochemical analysis of the samples, plus the characterization of druse crystals by energy-dispersive X-ray spectroscopy. The sample showed menthol and camphor odor, and bitter taste. The main macroscopic characteristic was the presence of a thin periderm with striations. The main distinctive microscopic features of the species were: dense gelatinous fibers, phenolic idioblasts, sclereids, numerous crystalline idioblasts with druses located in the cortex and abundantly in the parenchymal rays of the phloem, and absence of laticifers in the mature stem bark. These analyses made possible to describe the morpho-anatomy of that species, contributing to phytochemical and pharmacognostic future studies of C. floribundus.
... The tree shows rapid growth, and may be used in recovery planting (Lorenzi, 2002). The wood of some species of this genus have already been studied (Luchi, 2004; Wiedenhoeft, 2008; Wiedenhoeft et al., 2009; Longui et al., 2012; Novello et al., 2012; Siegloch et al., 2012). However, no studies in the literature have reported on C. piptocalyx wood. ...
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