Journal of Ethnopharmacology 141 (2012) 895–900

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Journal of Ethnopharmacology
journal homepage: www.elsevier.com/locate/jethpharm

Vasorelaxant action of the total alkaloid fraction obtained from Solanum

paludosum Moric. (Solanaceae) involves NO/cGMP/PKG pathway
and potassium channels
Fabio S. Monteiro a , Ana C.L. Silva a , Italo R.R. Martins a , Ana C.C. Correia a , Ionaldo J.L.D. Basílio a ,
Maria F. Agra a,c , Jnanabrata Bhattacharyya b , Bagnólia A. Silva a,c,∗
a
Centro de Ciências da Saúde/Universidade Federal da Paraíba, Brazil
b
University of Georgia, Department of Chemistry, Athens, United States
c
Departamento de Ciências Farmacêuticas, Centro de Ciências da Saúde, Universidade Federal da Paraíba, Brazil

a r t i c l e i n f o a b s t r a c t

Article history: Ethnopharmacological relevance: Solanum paludosum Moric. (jurubeba-roxa) is commonly used to treat
Received 2 December 2011 hypertension as a substitute for Solanum paniculatum L. (jurubeba verdadeira). The total ethanolic extract
Received in revised form 13 February 2012 from the root bark of Solanum paludosum have been found to cause hypotension in rats.
Accepted 17 March 2012
Aim of the study: To investigate the mechanism by which the total alkaloid fraction obtained from the
Available online 26 March 2012
root bark of Solanum paludosum (FAT-SP) acts as a vasorelaxant agent on rat thoracic aorta.
Materials and methods: Rings of rat aorta were suspended in organ bath containing Krebs solution at 37 ◦ C,
Keywords:
bubbled with carbogen mixture (95% O2 and 5% CO2 ) under a resting tension of 1 g. Isometric contractions
Solanum paludosum
Total alkaloids
were measured using a force transducer coupled to an amplifier and a microcomputer.
Rat aorta Results: FAT-SP has been found cause relaxation of the aortic rings pre-contracted with phenyle-
NO/sCG/PKG pathway phrine (Phe) in a concentration-dependent manner, in the presence and absence of endothelium.
Potassium channels This effect was more potent on the endothelium-intact aorta. In the presence of endothelium, nei-
ther indomethacin (non-selective cyclooxygenase inhibitor) nor atropine (non-selective muscarinic
receptor antagonist), produced significant changes on the relaxation response. On the other hand,
in the presence of calmidazolium (a calmodulin inhibitor), N-nitro-l-arginine methyl ester (l-NAME,
nitric oxide synthase inhibitor), hydroxocobalamin (HDX) (scavenger of free-radical nitric oxide), 1-
H-[1,2,4]-oxadiazolo-[4,3a]-quinoxalin-1-one (ODQ, selective blocker of soluble guanylate cyclase),
Rp-8-bromo-␤-phenyl-1,N2 -ethenoguanosine 3 :5 -cyclic monophosphorothioate sodium salt hydrate
(Rp-8-Br-PET-cGMPS, competitive inhibitor of cGMP-dependent protein kinase G) or TEA+ (tetraethy-
lammonium, nonselective potassium channel blocker), the vasorelaxant effect was significantly reduced,
suggesting the involvement of NO/sCG/PKG pathway and potassium channel opening in vasorelaxant
action of the FAT-SP.
Conclusion: The mechanism of vasorelaxant activity of the FAT-SP on rat aorta involves both NO/sCG/PKG
pathway and potassium channels.
© 2012 Elsevier Ireland Ltd. All rights reserved.

1. Introduction

The mechanisms of actions by which the plants can modulate

Abbreviations: FAT-SP, total alkaloid fraction of the root-bark of Solanum
paludosum; NOS, NO synthase; sGC, soluble guanylate cyclase; GTP, guanosine
triphosphate; cGMP, cyclic guanosine monophosphate; PKG, cGMP-dependent pro-
tein kinase G; Cav , L-type voltage-gated Ca2+ channels.
∗ Corresponding author at: Universidade Federal da Paraíba, Centro de
Biotecnologia/Pós-Graduação em Produtos Naturais e Sintéticos Bioat-
ivos/Laboratório de Farmacologia Funcional Prof. George Thomas, Cidade
Universitária, P.O. Box: 5009, Zip Code: 58051-970, João Pessoa, Paraíba, Brazil.
Tel.: +55 83 32167126; fax: +55 83 32167502.
E-mail addresses: bagnoliasilva@yahoo.com, bagnolia@ltf.ufpb.br (B.A. Silva).
NO action can be promising candidates for vasodilation, which may
have the potential to prevent and treat cardiovascular diseases such
as hypertension (Park et al., 2009).
In Brazil, several species of Solanum (S. paniculatum L., S. melon-
gena L. and S. stipulaceum Roem & Schult) were reported to induce
hypotension in rats (Ribeiro et al., 1986, 2002).
The systemic arterial hypertension is characterized by high
blood pressure (>140/90 mm Hg) and is a major risk factor responsi-
ble for the development of cardiovascular disease, renal failure and

0378-8741/$ – see front matter © 2012 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2012.03.032
896 F.S. Monteiro et al. / Journal of Ethnopharmacology 141 (2012) 895–900
endothelial dysfunction. Hypertension is one of the most important
public health problems in Brazil (Longo et al., 2009).
Antihypertensive drugs lower the blood pressure by acting on
the resistance vessels, the capacitance vessels, the heart, and the
kidney. Diuretics, sympatholytic drugs, vasodilators, calcium chan-
nel blockers, angiotensin-II blockers and angiotensin converting
enzyme inhibitors are important classes of antihypertensive drugs
(Webb et al., 2010).
Nitric oxide (NO) has been found to be, in recent years, an impor-
tant regulator of vascular functions by controlling blood vessel tone
as well as the interaction of blood cells with the vessel wall. The
NO/cGMP/PKG pathway and potassium channels play an essential
role in vascular smooth muscle relaxation, and the clinical studies
indicate that endothelium-derived NO is involved in normal and
pathological blood pressure regulation in humans (Moncada and
Higgs, 2006).
Solanum paludosum is a shrub commonly known as “jurubeba-
roxa” in northeastern Brazil (Agra and Bhattacharyya, 1999), and
the root bark of this plant is often used in folk medicine to
treat hypertension and other diseases as a substitute for Solanum
paniculatum (jurubeba-verdadeira) (Ribeiro et al., 1986).
In a recent study aimed to investigate the pharmacological
properties, the total alkaloid fraction of the root bark of Solanum
paludosum (FAT-SP) did not show any hemolytic activity on ery-
throcytes of rats and displayed spasmolytic activity on rat uterus
and guinea-pig trachea (Correia et al., 2011).
A chemical study of the FAT-SP resulted in the identification
(IR, 1 H and 13 C NMR with APT and DEPT) of the following com-
pounds: N-hydroxysolasodine, leptinidine, tomatidenol and a new
compound, putuline (see Bhattacharyya et al., 2009).
Moreover, the total ethanolic extract of the root bark of Solanum
paludosum has been found to have curare-like and hypotensive
activity in rats (Ataíde, “unpublished results”). Therefore, we inves-
tigated the vasorelaxant effect and action mechanism of the FAT-SP
on isolated rat aorta.
2. Methods
2.1. Plant material
The plant material of Solanum paludosum was collected in
November 2005 in the municipality of Santa Rita, State of Paraíba,
Brazil. The material was collected and identified by Maria de Fátima
Agra, and the voucher specimen (Agra 6734) is deposited at the
Prof. Lauro Pires Xavier (JPB) Herbarium, Universidade Federal da
Paraíba, João Pessoa, PB, Brazil.
2.2. Extraction of FAT-SP
The dried and powdered root bark (1.1 kg) of Solanum paludo-
sum was extracted with MeOH:H2 O (95:5) until the last extract
was free from color. The solvent was then removed under reduced
pressure, 5% H2 SO4 was added and the mixture was left for stand-
ing overnight. The product was then filtered through celite and
the filtrate was extracted with ether to remove fatty material. The
aqueous acid phase was then cooled, basified with NH4 OH to pH 9
and left standing for a few hours. The precipitated alkaloids were
collected by filtration which is designated FAT-SP (Scheme in Sup-
plementary Material).
2.3. Drugs
Magnesium sulfate heptahydrate (MgSO4 ·7H2 O), calcium chlo-
ride dihydrate (CaCl2 ·2H2 O), potassium chloride (KCl) and sodium
bicarbonate (NaHCO3 ) were obtained from Vetec (Brazil).
Tetraethylammonium chloride (TEA+ ), dimethyl sulfoxide
(DMSO), histamine hydrochloride, atropine sulfate, indomethacin,
NG -nitro-l-arginine methyl ester (l-NAME), phenylephrine
(Phe), 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ),
Rp-8-bromo-␤-phenyl-1,N2 -ethenoguanosine 3 :5 -cyclic
monophosphorothioate sodium salt hydrate (Rp-8-Br-PET-cGMPS)
and calmidazolium chloride were obtained from Sigma–Aldrich
(USA).
Glucose (C6 H12 O6 ) and monobasic potassium phosphate
(KH2 PO4 ·H2 O) were obtained from Nuclear (Brazil).
Sodium chloride (NaCl) and acetylcholine hydrochloride (ACh)
were obtained from Merck (Brazil).
Hydroxocobalamin hydrochloride (HDX) was obtained from
Bristol-Myers Squibb (Equador). Carbogen mixture (95% O2 and 5%
CO2 ) was obtained from White Martins (Brazil).
FAT-SP, provided by our collaborator of phytochemistry
(Bhattacharyya et al., 2009) was dissolved in distilled water to the
concentration of (10 mg/mL) and stored at 0 ◦ C. The stock solu-
tion was diluted in distilled water according to each experimental
protocol.
All substances were kept in freezer at −20 ◦ C except HDX which
was kept under room temperature. Indomethacin was dissolved
and diluted in a in a 0.2 M NaHCO3 solution and ODQ in DMSO.
2.4. Animals
Male Wistar rats (Rattus norvegicus), 12 weeks old and weighing
250–300 g each, obtained from Bioterium Prof. Thomas George of
UFPB, were used in this study. The animals were housed in cages
at a temperature of 21 ± 1 ◦ C in a 12 h light/12 h dark cycle and free
access to food and water. All procedures were approved by the UFPB
Animal Research Ethics Committee (protocol/CEPA no. 0111/09).
2.5. Tissue preparation
All rats were euthanized by decapitation with guillotine. The
aortic rings about 3–5 mm wide were obtained from the thoracic
aorta and were freed from the surrounding connective tissue. To
obtain isometric responses, the rings were individually suspended
on stainless steel rods in organ baths (6 mL) containing normal
Krebs solution at 37 ◦ C with the following composition (mM): NaCl
(118.0), NaHCO3 (25.0), KCl (4.6), MgSO4 (5.7), KH2 PO4 (1.1), CaCl2
(2.5) and glucose (11.0). The test solutions were stabilized for 1 h.
During this period, the aortic rings were kept under a resting ten-
sion of 1 g. In the meantime, the nutrient solution was renewed
every 15 min to prevent metabolite interference (Altura and Altura,
1970). In some of the rings, usual care was taken to avoid abrasion
of the inner surface and maintain the integrity of the endothelial
layer. In the remaining aortic rings, endothelial cells were removed
by gently rubbing the inner surface with moistened cotton swabs.
After the initial stabilization period, a contraction was induced
with Phe (0.3 ␮M) and when the tonic phase was reached
(12–15 min), ACh (1 ␮M) was added to all preparations to verify
endothelium integrity (Furchgott and Zawadzki, 1980). The vas-
cular endothelium was considered intact when the aortic rings
showed more than 50% of relaxation (Ajaya et al., 2003). Endothe-
lium removal was confirmed by the absence of relaxation after ACh
was added to the bath, or when the relaxation was lower than 10%,
when these rings were considered free of functional endothelium
(Fig. 1).
2.6. Experimental protocol
2.6.1. Effect of FAT-SP on Phe-induced tonic contractions in
endothelium-intact and -denuded rat aortic rings
Aortic rings were obtained as described in Section 2.5. After
the verification of endothelium integrity and during the tonic
 F.S. Monteiro et al. / Journal of Ethnopharmacology 141 (2012) 895–900
Fig. 1. Representative originals records in the absence (control, A) and presence of FAT-SP on endothelium-intact (B) and denuded (C) rat aorta rings. Arrow represents
concentration of FAT-SP (1, 3, 9, 27, 81, 243, 500 and 750 ␮g/mL). Phe: phenilefrine and W: washout.
component of a second response to the agonist, FAT-SP was cumu-
latively added to the bath in different preparations. The relaxation
was expressed as the reversal percentage of initial contraction
elicited by contractile agents and EC50 values (concentration of a
substance that produces 50% of its maximum effect) were obtained
with nonlinear regression from FAT-SP concentration–response
curves of rat aortic rings with both endothelium-intact and -
denuded aorta.
2.6.2. Effect of FAT-SP on Phe-induced tonic contractions in
endothelium-intact rat aortic rings in the presence of atropine or
indomethacin
The aorta was mounted as described in Section 2.5. Before
taking the second concentration–response curve induced by Phe,
the preparations with endothelium were separately incubated for
15 min with atropine (1 ␮M), a nonselective muscarinic receptor
antagonist (Barnes, 2000) or indomethacin (10 ␮M), a nonselective
inhibitor of cyclooxygenase (Sigthorsson et al., 2000). Even in the
presence of blockers, an additional Phe contraction was obtained.
Subsequently, FAT-SP was cumulatively added to the organ bath
and on the sustained component of contraction. The relaxation
was expressed as a percentage of the reverse contraction produced
by Phe. EC50 values were calculated by the nonlinear regression
method from the relaxation curves obtained both in the presence
and in the absence of blockers.
2.6.3. Effect of FAT-SP on Phe-induced tonic contractions in
endothelium-intact rat aortic rings in the presence of
calmidazolium, l-NAME, HDX, ODQ or Rp-8-Br-PET-cGMPS
The aorta was obtained as described in Section 2.5. Before
inducing the second Phe contraction in rings with functional
endothelium, the following agents were added in different prepa-
rations: l-NAME, a competitive NO synthase inhibitor (100 ␮M for
30 min) (Rees et al., 1990); calmidazolium, a calmodulin inhibitor
(10 ␮M for 30 min) (Schini and Vanhoutte, 1992), HDX, a NO rad-
ical scavenger (30 ␮M for 30 min) (Broderick et al., 2005); ODQ, a
897
selective soluble guanylyl cyclase (sCG) blocker (10 ␮M for 15 min)
(Garthwaite et al., 1995); and Rp-8-Br-PET-cGMPS, a competi-
tive cGMP-dependent protein kinase G (PKG) inhibitor (30 ␮M for
30 min) (Ibarra-Alvarado et al., 2002).
In the tonic component of the second Phe-induced contraction
in the presence of each blocker, FAT-SP was cumulatively added to
the bath in different preparations. The relaxation was expressed as
previously described.
2.6.4. Effect of FAT-SP on KCl-induced (30 or 80 mM) tonic
contractions in endothelium-intact rat aortic rings
The aortic rings were obtained as described in Section 2.5. After
washing the preparation and during the tonic component of a
second response achieved with KCl (30 or 80 mM) solution (Pérez-
Vizcaíno et al., 1998), FAT-SP was added cumulatively to the bath, in
different preparations. The relaxation was expressed as previously
described.
2.6.5. Effect of FAT-SP on Phe-induced tonic contractions in
endothelium-intact rat aortic rings in the presence of TEA+
The aorta was obtained as described in Section 2.5. Before
inducing the second Phe-induced contraction in rings with intact
endothelium, TEA+ (10 mM), a nonselective potassium channel
blocker (Sordi et al., 2011), was added in different preparations.
In the tonic component of the second Phe-induced contraction in
the presence of TEA+ , FAT-SP was cumulatively added to the bath in
different preparations. The relaxation was expressed as previously
described.
2.7. Statistical analysis
The values were expressed as mean ± S.E.M. Statistical analysis
was performed using Graph-Pad Prism 5.01 software (Graph-
Pad Software Inc., San Diego, CA, USA). The differences between
mean values were statistically compared using t-test and one-way
ANOVA, followed by Bonferroni correction when applicable, and
898 F.S. Monteiro et al. / Journal of Ethnopharmacology 141 (2012) 895–900

0 0

25 25

Relaxation (%)
Relaxation (%)

50 50
Phe

75 Endothelium-intact 75 Indomethacin

Endothelium-denuded
Atropine

100 100

0 1 2 3 0 1 2 3
log [FAT-SP] µg/mL log [FAT-SP] µg/mL
Fig. 2. Effect of FAT-SP on Phe-induced (0.3 ␮M) tonic contraction in rat aorta rings Fig. 3. Effect of FAT-SP on Phe-induced (0.3 ␮M) tonic contraction in endothelium-
with intact (䊉) or denuded () endothelium. Symbols and vertical bars represent intact rat aorta rings in absence (control, 䊉) and presence of atropine (1 ␮M, )
means and S.E.M., t-test, ***p < 0.001 (endothelium-intact vs. endothelium-denuded) or indomethacin (10 ␮M, ). Symbols and vertical bars represent means and S.E.M
(n = 5). (n = 5).

were considered to differ significantly when p < 0.05. The EC50 val-
ues were determined using nonlinear regression (Neubig et al.,
2003).
3. Results and discussion
This study demonstrated for the first time that the total alka-
loid fraction obtained from the root bark of Solanum paludosum
(FAT-SP) shows vasorelaxant effect in isolated rat aorta. Moreover,
the most significant finding concerns the characterization of the
vasorelaxant action mechanism, which includes the activation of
NO/sCG/PKG pathway and the potassium channels. The presence
of steroid alkaloids may be responsible for the vasorelaxant effect
of FAT-SP.
The FAT-SP relaxed rat aorta pre-contracted with Phe
(0.3 ␮M) in a concentration-dependent manner on both intact
(EC50 = 75.4 ± 6.2 ␮g/mL) and denuded (EC50 = 242.8 ± 11.7 ␮g/mL)
endothelium (Fig. 2). The fact that vasorelaxant effect of the FAT-SP
was more potent on endothelium-intact rat aorta is suggestive of
the involvement of endothelium-derived relaxing factors (EDRF)
such as NO and prostacyclin (PGI2 ). The Phe-induced contraction
was reversible 30 min after the withdrawal of FAT-SP from the bath
calmidazolium, a calmodulin inhibitor to investigate a possible
calmodulin role in the relaxation produced by FAT-SP on rat aorta.
Since this effect produced by FAT-SP was attenuated with the
decrease in the relaxation potency (EC50 = 379.0 ± 64.8 ␮g/mL) and
maximum effect reduction (Fig. 4) in the presence of calmida-
zolium, it appears that vasorelaxant effect of the FAT-SP involves
calmodulin participation.
The endothelium-derived NO is more likely free radical (NO• )
form (Feelisch et al., 1994). Therefore, we used HDX, a NO•
scavenger, to investigate NO role in the relaxation produced
by FAT-SP on rat aorta. The relaxation potency and effect
(EC50 = 294.9 ± 20.4 ␮g/mL) of FAT-SP was reduced (Fig. 4) in the
presence of HDX, it is suggestive that vasorelaxant effect of the
FAT-SP involves NO participation.
Solanum species are known to be rich in steroid alkaloids and
several species present relevant hypotensive activities. It has been
reported in the literature that substances with a steroidal skeleton
500
**
400
***
EC50 (µg/mL)

(data not shown).

300 ***
Thus, we investigated the role of muscarinic receptors,
** ***
cyclooxygenase enzyme and NO synthase (eNOS) using atropine,
indomethacin and l-NAME, respectively. 200
The relaxation induced by FAT-SP (EC50 = 75.4 ± 6.2 ␮g/mL)
on endothelium-intact rat aortic rings pre-contracted with 100
Phe was not significantly attenuated in the presence of
both atropine (EC50 = 93.3 ± 10.0 ␮g/mL), or indomethacin 0
(EC50 = 90.7 ± 13.2 ␮g/mL) (Fig. 3). However, the vasorelaxant
m

Q
ol

PS
E

effect was found to be attenuated, as observed in the EC50 values

iu
tr

M
H

O
on

ol

cG

(EC50 = 335.8 ± 32.5), in Fig. 4, and maximum effect reduction

N

az
C

L-

T-
id

in the presence of l-NAME, indicating eNOS involvement in the

PE
m
al

r-

vasorelaxant effect of the FAT-SP.

C

B
8-

The participation of eNOS was also detected in other Solanum

p-
R

species, e.g., Solanum indicum ssp. distichum and Solanum torvum

Swartz which contain steroidal alkaloids (Bahgat et al., 2008;
Nguelefack et al., 2009).
The eNOS is activated by calcium–calmodulin (Ca2+ /CaM)
complex which binds to NOS in response to increases in intra-
cellular calcium concentration (Stuehr, 1997). Therefore, we used
[FAT-SP µg/mL]
Fig. 4. FAT-SP EC50 values in absence and presence of the blockers l-NAME
(100 ␮M), calmidazolium (10 ␮M), HDX (30 ␮M), ODQ (10 ␮M) or Rp-8-Br-PET-
cGMPS (30 ␮M). Columns and bars represent means and S.E.M. One-way ANOVA
followed by Bonferroni’s test, **p < 0.01, ***p < 0.001 (n = 5).
 F.S. Monteiro et al. / Journal of Ethnopharmacology 141 (2012) 895–900
0 0
25
Relaxation (%)
50
KCl 30 mM
75
KCl 80 mM
100
0 1 2 3
log [FAT-SP] µg/mL
Fig. 5. Effect of FAT-SP on tonic contraction induced by KCl (30 mM, ) and KCl
(80 mM, ) in endothelium-intact rat aorta rings. Symbols and vertical bars repre-
sent means and S.E.M (n = 5).
show vasorelaxant activity in an endothelium-dependent, NO-
mediated manner in rat aorta (Achike and Kwan, 2003), supporting
our results.
The soluble guanylate cyclase (sCG) is a key protein in NO/cGMP
signaling pathway (Sharina et al., 2008). Therefore, we investi-
gated the vasorelaxation effect of the FAT-SP in the presence
of ODQ. As a result, we observed that vasorelaxation effect and
potency of the FAT-SP were reduced (Fig. 4) in the presence of ODQ
(EC50 = 228.2 ± 30.9 ␮g/mL), indicating the participation of sGC.
sGC converts guanosine triphosphate (GTP) into the second
messenger 3 :5 -cGMP (Sharina et al., 2008). An increase in cGMP
concentration results in the PKG activation which phosphorylates
specific targets, e.g., potassium channels. This leads to their open-
ing with hyperpolarization of membrane and blockade of L-type
voltage-gated Ca2+ channels (CaV ), which causes the reduced cyto-
plasmic Ca2+ concentration and, consequently, vascular smooth
muscle relaxation (Hofmann et al., 2000). Thus, the PKG com-
petitive inhibitor Rp-8-Br-PET-cGMPS was used. The relaxation
potency of the FAT-SP was reduced (EC50 = 253.7 ± 18.7 ␮g/mL)
(Fig. 4), showing a PKG participation in the vasorelaxant effect of
FAT-SP on isolated rat aorta.
Since both NO and PKG may directly activate potassium chan-
nels in the smooth muscle (Bolotina et al., 1994) to produce
relaxation and PKG can inhibit L-type CaV , we investigated this
hypothesis with a simple protocol using elevated extracellular
potassium [K+ ]e for a better characterization of the NO/cGMP/PKG
pathway.
The FAT-SP significantly relaxed rat aorta pre-contracted
with KCl (30 mM) in a concentration-dependent manner on
endothelium-intact (EC50 = 98.8 ± 26.5 ␮g/mL). In KCl-induced
contractions (80 mM), the FAT-SP showed maximum effect of
10.2 ± 2.3% on endothelium-intact rat aortic rings (Fig. 5).
K+ channel openers effectively relax vascular smooth muscles
under moderate [K+ ]e conditions, but this effect was markedly
weakened at higher [K+ ]e concentrations (Pérez-Vizcaíno et al.,
1998). The present findings demonstrate that FAT-SP relaxes
endothelium-intact rat aorta more effectively when pre-contracted
with 30 mM of KCl than with a higher amount (80 mM) of KCl, sug-
gesting that the vasorelaxant effect of the FAT-SP on rat aorta is
due to a positive modulation of K+ channel and indirect L-type CaV
closing.
899
25
Relaxantion (%)
50
Phe
75
TEA+
100
0 1 2 3
log [FAT-SP] µg/mL
Fig. 6. Effect of FAT-SP on Phe-induced (0.3 ␮M) tonic contraction in endothelium-
intact rat aorta rings in absence (control, 䊉) and presence of presence of TEA+
(10 mM, ). Symbols and vertical bars represent means and S.E.M., t-test, ***p < 0.001
(control vs. TEA+) (n = 5).
In order to confirm the hypothesis of K+ channel involve-
ment in vasorelaxant effect of the FAT-SP, experiments were
performed using endothelium-intact rat aorta in the presence of
TEA+ (10 mM). The relaxation curve of the FAT-SP was shifted to
the right in the TEA+ 10 mM presence, reducing the relaxation
potency (EC50 = 217.3 ± 39.6 ␮g/mL), which suggests potassium
channel involvement in the vasorelaxant effect of the FAT-SP
(Fig. 6).
There is currently a growing interest in the therapeutic poten-
tial of substances that modulate potassium channels. There are
few compounds potassium channel activators; cassiarin A being
an example of an alkaloid among those (Matsumoto et al., 2010).
In conclusion, the vasorelaxant action of the total alkaloids frac-
tion from root barks of Solanum paludosum (Solanaceae) (FAT-SP)
on rat aorta involves NO/sCG/PKG pathway and potassium channel
opening. Further complementary studies on this alkaloid fraction
using molecular experiments and in vivo studies are needed to pur-
sue the search for a potential antihypertensive agent.
Acknowledgements
The authors thank Jose Crispim Duarte (Laboratory Technician/
UFPB), CNPq and CAPES (organ development).
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.jep.2012.03.032.
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