ArticlePDF Available

Abstract

Background: Salvia urica Epling is taxonomically and phylogenetically related to Salvia amarissima Ortega. The last species has pharmacological relevance by its contents of bioactive metabolites. Nowadays, Salvia urica has no reports about its chemical constituents and pharmacological activities. Hypothesis: Does the close relationship between S. amarissima and S. urica led both species produce similar specialized metabolites? Does Salvia urica display similar pharmacological effects as S. amarissima? Studied species: Salvia urica Epling (Lamiaceae). Study site and dates: The plant material was collected in Teopisca, Chiapas, Mexico, in December 2021. Methods: Metabolites of the acetone extract from Salvia urica were identified by GC-MS and HPLC-PDA profiling. In parallel, a phytochemical study was conducted, and the individual constituents purified, previously characterized by 1D NMR, were assayed on antihyperglycemic effect in diabetic mice and a charcoal-gum arabic-induced hyperperistalsis model in rats. Results: The volatile compounds identified by GC-MS were alkanes, aromatics and triterpenes. The principal constituents of the acetone extract of Salvia urica were amarissinin A and 5,6-dihydroxy-7,3',4'-trimethoxyflavone, which were also quantified by HPLC-PDA. The extract and both metabolites isolated showed an antihyperglycemic effect on streptozotocin-induced diabetic mice, suggesting a possible synergic effect. In addition, the compound 5,6-dihydroxy-7,3',4'-trimethoxyflavone (IC50 = 0.79 mg/kg) showed a better antipropulsive effect than loperamide (IC50 = 16.6 mg/kg). Conclusions: The phytochemical composition of an acetone extract of Salvia urica was determined by first time. The metabolites isolated from this plant support the phylogenetic relationship of S. urica with Salvia amarissima, and they showed antipropulsive and antihyperglycemic effects.
1
García-Nava et al. / Botanical Sciences
ChemiCal Constituents of Salvia urica epling, and their antihyperglyCemiC
and antipropulsive effeCts
XitlaliCk garCía-nava1,2, miguel valdes3,4, fernando Calzada4, elihú Bautista2*,
omar Cortezano-arellano5, denisse de loera1, itzi fragoso-martínez6 and martha martínez-gordillo7
1 Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, San Luis Potosí, Mexico.
2 CONAHCYT-División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica A. C., San Luis Potosí,
Mexico.
3 Instituto Politécnico Nacional, Sección de Estudios de Posgrado e Investigación, Escuela Superior de Medicina, Mexico City, Mexico.
4 Unidad de Investigación Médica en Farmacología, UMAE Hospital de Especialidades, Centro Médico Nacional Siglo XXI, IMSS,
Mexico City, Mexico
5 Instituto de Ciencias Básicas, Universidad Veracruzana, Xalapa, Veracruz, Mexico.
6 Red de Biodiversidad y Sistemática, Instituto de Ecología, A.C., Xalapa, Veracruz, México.
7 Herbario de la Facultad de Ciencias (FCME), Universidad Nacional Autónoma de México, Mexico City, Mexico.
*Corresponding author: francisco.bautista@ipicyt.edu.mx
Botanical Sciences
DOI: 10.17129/botsci.3368
Received: June 22, 2023, Accepted: October 4, 2023
On line rst: November 27, 2023
Phytochemistry / Fitoquímica
This is an open access article distributed under the terms of the Creative Commons Attribution License CCBY-NC (4.0) international.
https://creativecommons.org/licenses/by-nc/4.0/
Abstract
Background: Salvia urica Epling is taxonomically and phylogenetically related to Salvia amarissima Ortega. The last species has pharmaco-
logical relevance by its contents of bioactive metabolites. Nowadays, Salvia urica has no reports about its chemical constituents and pharma-
cological activities.
Hypothesis: Does the close relationship between S. amarissima and S. urica led both species produce similar specialized metabolites? Does
Salvia urica display similar pharmacological effects as S. amarissima?
Studied species: Salvia urica Epling (Lamiaceae).
Study site and dates: The plant material was collected in Teopisca, Chiapas, Mexico, in December 2021.
Methods: Metabolites of the acetone extract from Salvia urica were identied by GC-MS and HPLC-PDA proling. In parallel, a phytochemi-
cal study was conducted, and the individual constituents puried, previously characterized by 1D NMR, were assayed on antihyperglycemic
effect in diabetic mice and a charcoal-gum arabic-induced hyperperistalsis model in rats.
Results: The volatile compounds identied by GC-MS were alkanes, aromatics and triterpenes. The principal constituents of the acetone extract
of Salvia urica were amarissinin A and 5,6-dihydroxy-7,3’,4’-trimethoxyavone, which were also quantied by HPLC-PDA. The extract and
both metabolites isolated showed an antihyperglycemic effect on streptozotocin-induced diabetic mice, suggesting a possible synergic effect.
In addition, the compound 5,6-dihydroxy-7,3’,4’-trimethoxyavone (IC50 = 0.79 mg/kg) showed a better antipropulsive effect than loperamide
(IC50 = 16.6 mg/kg).
Conclusions: The phytochemical composition of an acetone extract of Salvia urica was determined by rst time. The metabolites isolated from
this plant support the phylogenetic relationship of S. urica with Salvia amarissima, and they showed antipropulsive and antihyperglycemic effects.
Keywords: Specialized metabolites, terpenoids, Salvia genus, sage, antidiarrheal, antidiabetic, chromatographic methods, NMR analysis.
Resumen
Antecedentes: Salvia urica Epling está relacionada taxonómica y logenéticamente con Salvia amarissima Ortega. Ésta última tiene relevancia
farmacológica por su contenido de metabolitos secundarios. A la fecha, Salvia urica no cuenta con reportes de sus constituyentes químicos y de
sus actividades farmacológicas.
Hipótesis: ¿La relación cercana entre S. amarissima y S. urica permite que ambas especies produzcan metabolitos similares de tipo terpenoide?
¿Salvia urica produce un efecto farmacológico similar a S. amarissima?
Especies de estudio: Salvia urica Epling (Lamiaceae)
Sitio y años de estudio: El material vegetal fue colectado en Teopisca, Chiapas, México, en diciembre de 2021.
Métodos: Los constituyentes de las partes aéreas de Salvia urica, se identicaron por medio del perlamiento por GC-MS y HPLC-PDA. Para-
lelamente, se realizó un estudio toquímico, y los constituyentes individuales, caracterizados por RMN de 1D, se evaluaron en un ensayo de
actividad antihiperglucémica y en un modelo de hiperperistalsis.
Botanical Sciences / Online First
Botanical Sciences / Online First
Chemical constituents of Salvia urica
2
Resultados: Los componentes volátiles identicados por GC-MS son de tipo alcano, aromático y triterpenoide. Los principales constitu-
yentes no volátiles, son amarissinina A y 5,6-dihidroxi-7,3’,4’-trimetoxiavona. Ambos metabolitos se cuanticaron por HPLC-PDA. El
extracto de acetona y los dos metabolitos aislados mostraron efecto antihiperglucémico en ratones diabéticos, sugiriendo que hay un posible
efecto sinérgico. El compuesto 5,6-dihidroxi-7,3’,4’-trimetoxiavona (IC50 = 0.79 mg/kg) mostró un mejor efecto antipropulsivo que lopera-
mida (IC50 = 16.6 mg/kg).
Conclusiones: Se identicaron por primera vez los metabolitos mayoritarios de Salvia urica. Dos de estos metabolitos se aislaron previamente
de Salvia amarissima, apoyando su relación logénetica. Los dos compuestos mostraron efecto antipropulsivo y antihiperlgucémico.
Palabras clave: Metabolitos especializados, terpenoides, género Salvia, antidiarreico, antidiabético, métodos cromatográcos, análisis por
RMN
The Lamiaceae family contains 7,139 species (Harley et al. 2005), and the genus Salvia groups nearly
1,000 of them. From these species, 306 are distributed in Mexico. Salvia urica Epling is a perennial
herbaceous plant native to Mexico, with a restricted distribution in the state of Chiapas, as well as in
Guatemala and Belize (Martínez-Gordillo et al. 2017). It grows at elevations from 400 to 2,600 meters
above mean sea level, in seasons ranging from summer to winter (Clebsch 2003). This plant is commonly known as
“pendolita morada”, “canastillas”, “tutzunún”, “Salvia bretónica”, and “chichingua azul” (Chicago Natural History
Museum 1973, Padilla-Gómez 2007). Some traditional medicinal uses given to S. urica are to treat “bilis”, diabetes
mellitus, stomachache, and diarrhea (Padilla-Gómez 2007). However, no phytochemical and pharmacological re-
ports of the constituents derived from this plant species support its ethnomedical uses in Mexico. To date, there is just
a report describing the antibacterial effect of a methanolic extract of the plant against Escherichia coli (Sakagami et
al. 2001). Originally, Salvia urica and Salvia amarissima Ortega were classied together in section Uricae Epling,
due to their morphological resemblance (Epling 1939). Later, they were moved to a broader section: Scorodonia
Epling (Epling 1941). Phylogenetic studies of Salvia subgenus Calosphace have shown that both these sections are
articial groupings (Fragoso-Martínez et al. 2018). However, Salvia amarissima and S. urica are closely related,
forming a clade with Salvia leucochlamys Epling, Salvia ozolotepecensis J.G.González & Fragoso, Salvia perlonga
Fernald, and Salvia praestans Epling (González-Gallegos et al. 2019). The lack of phytochemical and pharmacologi-
cal studies, and the phylogenetic closeness between S. urica and S. amarissima prompted us to study the rst species;
in order to explore if it produces secondary metabolites of diterpenoid-type that supports this relationship since a
chemotaxonomic approach, and if it shares pharmacological properties with S. amarissima.
As part of our ongoing efforts to study the phytochemical composition of Mexican medicinal plants for drug
discovery and agrochemical purposes (Ortega et al. 2020, Bautista et al. 2022, García-Nava et al. 2022), herein, we
describe the isolation and identication of the major metabolites from an acetone extract of Salvia urica, as well as
the subsequent determination of the anti-hyperglycemic and anti-propulsive effects of these constituents.
Materials and methods
General experimental procedures. 1D NMR experiments were performed on an Agilent Mercury NMR spectrometer
500 MHz, equipped with a gradient probe for variable temperature experiments. Chemical shifts were referred to
TMS, and J values are given in Hertz (Hz). Column chromatography (CC) was performed on silica gel 60 (Merck G).
Thin-Layer Chromatography (TLC) was carried out on precoated Macherey-Nagel Sil G/UV254 plates of 0.25 mm
thickness, and spots were visualized by spraying with 3 % Ce(SO4)2 in H2SO4 2 N, followed by heating. The GC-MS
analyses were obtained on an Agilent gas chromatograph, model GC-7890b coupled to an Agilent mass spectrometer
EM-5877A, using a GC capillary column DB 5ht (30 m × 0.320 mm i.d. × 0.1 μm, Agilent). The HPLC analyses were
obtained on a 1200 HPLC Agilent chromatograph equipped with a G1315D Agilent PDA detector and a G1315A Agi-
lent uorescence detector. HPLC separation was performed in a Synergi Polar RP column (Phenomenex, 5 mm, 4.6 ×
250 mm) with a gradient of acetic acid 1 % (A) and acetonitrile (B) at the following times: 0 min, 80 % A; 1 min, 80 %
A; 31 min, 40 % A, 33 min, 20 % A; 43 min, 20 % A. The chromatograms were obtained at 30 °C; the standards used
Botanical Sciences / Online First
Botanical Sciences / Online First
3
García-Nava et al. / Botanical Sciences
were amarissinin A (1) (> 98 %, HPLC-PDA) and 5,6-dihydroxy-7,3’,4’-trimethoxyavone (2) (> 98 %, HPLC-PDA);
and the calibration curves of the standards were made using different concentrations 0.25-0.00625 mg/mL.
Plant material. The aerial parts were collected at the owering stage in Teopisca, Chiapas, Mexico, in December
2021. A voucher specimen was deposited at the Herbarium of Facultad de Ciencias, UNAM (FCME-180582), and
determined by Dr. Martha Martínez Gordillo.
Extraction and isolation. The dried and grounded aerial parts of Salvia urica (218 g) were subjected to extraction
by maceration with acetone (3 × 2 L). The extract was ltered, and the solvent was recovered by distillation at re-
duced pressure to obtain 11 g of dried residue. The S. urica acetone extract (SUE) was submitted to silica gel CC
(8.5 × 4.5 cm, 125 mL), eluting with hexane-EtOAc, EtOAc, and EtOAc-MeOH mixtures in increasing polarity. The
grouping fractions was conducted by their likeness observed in the TLC analyses as follows: Fraction A (frs.1-7,
eluted with hexanes-EtOAc 9:1), fraction B (frs. 8-22, eluted with hexanes-EtOAc 8:2), fraction C (frs. 23-27, eluted
with hexanes-EtOAc 3:2), fraction D (frs. 28-38, eluted with hexanes-EtOAc 3:2), fraction E (frs. 39-43, eluted with
hexanes- EtOAc 1:1), fraction F (frs. 44-52, eluted with hexanes-EtOAc 3:7), fraction G (fr. 53, eluted with EtOAc),
fraction H (fr. 54, eluted with EtOAc), fraction I (frs. 55-58, eluted with EtOAc-MeOH 9:1). Fraction D (1.6 g) was
submitted to silica gel CC (10.0 × 2.0 cm, 25 mL), eluting with CHCl3-MeOH mixtures in increasing polarity to give
ve fractions (D1-D5). Fr. D2 (eluted with CHCl3-MeOH 97:3) yielded 62 mg of amarissinin A (1) by sequential
crystallizations from acetone/hexanes, EtOAc/hexanes, and acetone/EtOAc. Fr. E yielded 7 mg of 5,6-dihydroxy-
7,3’,4’-trimethoxyavone (2), which was ltrated, and crystallized from EtOAc/hexanes.
Compound 1.- yellowish crystals, mp 219-224 °C; 1H NMR (500 MHz, CDCl3-DMSO-d6, Figures S1 and S2): δH
7.82 (br s, H-16), 7.39 (t, J = 1.8 Hz, H-15), 6.69 (ddd, J = 10.1, 4.9, 2.2 Hz, H-2), 6.54 (br d, J = 1.8 Hz, H-14), 6.12
(br s, H-11), 5.92 (dt, J = 10.1, 1.4 Hz, H-1), 4.72 (dd, J = 8.7, 1.5 Hz, H-19Pro R), 4.35 (d, J = 8.7 Hz, H-19Pro S), 2.71
(dd, J = 19.3, 4.9 Hz, H-3a), 2.61 (dt, J = 19.3, 2.2 Hz, H-3b), 2.47 (ddd, J = 13.1, 12.0, 5.1 Hz, H-7a), 2.21 (ddd, J
= 13.5, 11.6, 5.4 Hz, H-7b), 2.03 (s, H3-20), 1.61 (m, H-6a), 1.56 (m, H-6b); 13C NMR (125 MHz, DMSO-d6, Figure
S3): δC 197.39 (C, C-10), 173.89 (C, C-18), 161.75 (C, C-17), 151.47 (C, C-12), 150.37 (C, C-9), 143.96 (CH, C-15),
143.04 (CH, C-2), 141.48 (CH, C-16), 127.39 (CH, C-1), 120.72 (C, C-8), 119.31 (C, C-13), 106.62 (CH, C-14),
104.60 (CH, C-11), 75.81 (C, C-4), 69.49 (CH2, C-19), 57.18 (C, C-5), 29.84 (CH2, C-3), 28.79 (CH2, C-6), 21.95
(CH2, C-7) and 18.55(CH3, C-20).
Compound 2.- yellowish crystals, mp 240-241 °C; 1H NMR (500 MHz, DMSO-d6, Figure S4): δH 12.62 (s, OH-5),
8.72 (s, OH-6), 7.70 (d, J = 8.5 Hz, H-5’), 7.59 (br s, H-2’), 7.13 (dd, J = 8.5, 1.6 Hz, H-6’), 6.98 (s, H-8), 6.94 (s,
H-3), 3.88 (OCH3-7), 3.85 (OCH3-3’), 3.80 (OCH3-4’); 13C NMR (125 MHz, DMSO-d6, Figure S5): δC 182.26 (C,
C-4), 163.34 (C, C-2), 154.41 (C, C-7), 152.04 (C, C-4’), 149.03 (C, C-3’), 146.16 (C, C-5), 129.98 (C, C-6), 123.07
(C, C-1’), 119.98 (CH, C-6’), 111.7 (CH, C-5’), 109.39 (CH, C-2’), 103.45 (CH, C-3), 91.26 (CH, C-8), 56.16 (CH3,
OCH3-7), 55.73 (CH3, OCH3-3’), 55.58 (CH3, OCH3-4’).
Identication of the volatile constituents by GC-MS. A sample of 5 g of plant material was macerated with acetone
(3 × 60 mL) for 48 h to obtain residues ranging from 490 to 503 ± 9.9 mg (5.5 ± 0.08 % yield dry weight). Then, 10 mg
of extract were dissolved in 2 mL of acetone using sonication, and a volume of 0.2 μL containing this solution was
injected into the gas chromatograph (splitless). The temperature of the port injection was maintained at 220 °C, and
the oven was 80 °C for 2 min, hereafter increased at 3, 5, and 10 °C/min until 100, 150, and 330 °C, respectively.
The nal temperature was maintained for 9 min. Each analysis was repeated in triplicate, and the volatile com-
pounds were identied by deconvolution, using the W10N11 database (Wiley10Nist11) and the % area normalized
(Sepúlveda-Cuellar et al. 2021).
Botanical Sciences / Online First
Botanical Sciences / Online First
Chemical constituents of Salvia urica
4
High Performance Liquid Chromatography. A sample of plant material (10 g) was macerated with acetone (3 × 60 mL)
for 48 h. This procedure was repeated with three independent samples to obtain residues between 490–503 ± 9.9 mg.
Then, a solution of each extract at 2.5 mg/mL was prepared in methanol. The injection volume of standards was 5 μL,
and 10 μL for samples, using a ow rate of 0.8 mL/min. The temperature of the equipment was 30 °C. The PDA detector
acquires the chromatograms at 240 and 330 nm, and the FL detector operates with an excitation longwave of 250 nm, and
an emission longwave of 410 nm. All experiments were repeated in triplicate according to Sepúlveda-Cuellar et al. (2021).
Animals. Balb/c male mice of 8-10 weeks of age weighing 25 ± 5 g, and with glucose level values of 161 ± 5 mg/mL
were used for the antihyperglycemic test. Sprague-Dawley male rats (200-250 g) for the antipropulsive test were
used. The animals were obtained and raised in the Animal House of the National Medical Center “Siglo XXI” at In-
stituto Mexicano del Seguro Social (IMSS). The in vivo experiments were conducted following the Ofcial Mexican
NOM-0062-ZOO-1999 (SEMARNAT 1999) for Animal Experimentation and Care. The room temperature was 22 ±
2 °C with a 12-h light–dark natural cycle to maintain the animal care. The animals were fed with a standard diet and
water ad libitum. All in vivo experiments were conducted with the approval of the Specialty Hospital Ethical Com-
mittee of the National Medical Center “Siglo XXI” at IMSS (register: R-2019-3601-004).
Induction of experimental type 2 diabetes in mice. The diabetes was induced in male Balb/c mice using the strepto-
zotocin/nicotinamide model, according to a minor modication of the method described by Valdés et al. 2019. The
mice fasted for 16 h before being treated with an intraperitoneal solution of streptozotocin (100 mg/kg). After 30 min
of the streptozotocin administration, the mice were treated with nicotinamide (240 mg/kg in cold saline solution) via
intraperitoneal. The mice were fed with sucrose solution (10 %) ad libitum over three days at the end of the third day.
The sucrose solution was retired and substituted with water on the fth day. After 24 h, blood glucose levels were
measured by the glucose oxidase method (ACCU-CHECK® Instant Blood Glucose System, Roche, DC®, Mexico).
Antihyperglycemic effect. Balb/c mice were randomly divided into 8 groups (n = 6 per group). The groups were
separated by treatment as follows: Normoglycemic (NM control) and Diabetic (SID2 control) mice treated with
vehicle (2 % Tween 20 in water); NM mice and SID2 treated with SUE (300 mg/kg), compound 1 (50 mg/kg) and
compound 2 (50 mg/kg), respectively. The collection of blood samples from the tail vein was at 0, 2, and 4 h. The
analysis of blood samples was done by the glucose oxidase method (Valdés et al. 2019). The results were expressed
as mean values ± standard error of the mean (SEM). The statistical analyses were performed by GraphPad Prism
(GraphPad Software Inc., San Diego, CA, USA). The statistical evaluation was conducted by Bonferroni test for
multiple comparisons with a P < 0.05 of signicance.
Antipropulsive effect. The rats were fasted for 12 h before starting the experiment, but with water access ad libitum.
The method reported by Calzada et al. (2010) was followed with minor modications. The groups were divided into
control and test groups (n = 6 per group). The control group was treated with vehicle (1 mL 2 % DMSO in water) or
loperamide hydrochloride (10 mg/kg, 1 mL in 2 % DMSO, positive control), and the test groups were divided into
SUE (12.5 - 50 mg/kg), compound 1 (0.125 - 1.5 mg/kg) and compound 2 (0.125 - 1.5 mg/kg). After 20 min, the ani-
mals were administered with 1 mL of charcoal meal [10 % charcoal suspension in 5 % aqueous gum Arabic] by oral
route. After 30 min, the animals were sacriced, and their stomach and small intestine were removed and extended on
a glass surface. The distance from the pylorus to the caecum was measured and expressed as a percentage. All results
were expressed as mean ± S.E.M. and evaluated by Student’s t-test with a P < 0.05 of signicance.
Results
The GC-MS analysis of the volatile constituents from the acetone extract of Salvia urica indicated the presence of
16 metabolites, which include six triterpenes, seven alkanes and three aromatic metabolites such as anthraquinone
and naphthol derivatives (Supplementary material, Table S1). The relative amounts of the volatile compounds, by
Botanical Sciences / Online First
Botanical Sciences / Online First
5
García-Nava et al. / Botanical Sciences
class, were: 76.61, 15.39, and 7.99 %, respectively. The HPLC-PDA proling of the non-volatile constituents in the
acetone extract of S. urica, followed by the subsequent quantication of the isolated compounds (Supplementary
material, Figures S1-S5) by chromatographic methods, indicated that amarissinin A (1, rt = 24.24 min, 330 nm) and
5,6-dihydroxy-7,3’,4’-trimethoxyavone (2, rt = 24.58 min, 330 nm) are the major constituents in the extract (Figure
2, Table 1).
Figure 2. (A) HPLC-PDA proling chromatogram of amarissinin A (1). (B) HPLC-PDA proling chromatogram of 5,6-dihydroxy-7,3’,4’-trimethoxy-
avone (2). (C) HPLC-PDA proling chromatogram of the acetone extract of Salvia urica at 330 nm.
Figure 1. Chemical structures of amarissinin A (1) and 5,6-dihydroxy-7,3’,4’-trimethoxyavone (2).
In the normoglycemic mice, the extract and the metabolites isolated from Salvia urica did not signicantly affect
glucose levels (Table 2). However, in streptozotocin-induced type 2 diabetes mice, the extract and the metabolites
showed an antihyperglycemic effect at 2 and 4 h after their administration (Table 2). Compound 2 showed a dose-
dependent decrement in glucose levels; and, the acetone extract showed the lowest glucose level after 4 h of treat-
ment. A synergic effect between the constituents of the extract is possible.
Botanical Sciences / Online First
Botanical Sciences / Online First
Chemical constituents of Salvia urica
6
On the charcoal-gum Arabic-induced hyperperistalsis model in rats, the extract, and their major metabolites of
Salvia urica were tested at different doses. All showed a dose-dependence effect. Among them, amarissinin A (1)
showed the best inhibitory effect on the peristalsis at 0.125 mg/kg, and the 5,6-dihydroxy-7,3’,4’-trimethoxyavone
(2) showed a half maximal effective concentration of 0.79 mg/kg, in comparison with the positive control (1.003 mg/
kg, Table 3).
Acetone extract (yield in % w/w) Compound 1 (mg/g extract) Compound 2 (mg/g extract)
5.17 ± 0.09 163.36 ± 3.17 141.98 ± 4.59
Table 1. Quantication of amarissinin A (1) and 5,6-dihydroxy-7,3’,4’-trimethoxyavone (2) by HPLC-DAD (at 330 nm) in the total
extract from Salvia urica.
Note: Data expressed as average ± SD.
Treatment
Blood glucose levels (mg/dL)
0h 2h 4h
NM control 137.5 ± 5.6 131.3 ± 3 133 ± 3.5
SID2 control 300 ± 32.7 356.3 ± 21.5 351.3 ± 25.2
SID2 + SUE (300 mg/kg) 308.2 ± 7.6 285.2 ± 19 173 ± 31.7*, b
SID2 + 1 (50 mg/kg) 297.3 ± 6.9 337.8 ± 32.6 302.3 ± 45.4
SID2 + 2 (50 mg/kg) 289.8 ± 8.5 252.3 ± 5.4*, a237.3 ± 5.6*, b
NM + SUE (300 mg/kg) 134.5 ± 3.7 128.3 ± 5.2 134.5 ± 3.1
NM + 1 (50 mg/kg) 136 ± 4.9 136.5 ± 5.3 133.8 ± 5
NM + 2 (50 mg/kg) 134.8 ± 2.5 126.8 ± 2.2 130.3 ± 2
Table 2. Blood glucose levels of male normoglycemic mice (NM) and streptozotocin-induced type 2 diabetes mice (SID2) at 0, 2 and
4 h, on the acute antihyperglycemic test.
Discussion
The species of the Salvia genus have broad traditional medicinal uses in Mexico, which include treating digestive and
gynecological problems, and affections of the nervous system (Jenks & Kim 2013). Previous reports have explored
the phylogenetic relationships of species from Salvia subgenus Calosphace based on nuclear and plastid markers
(ITS, trnL-trnF, and trnH-psbA) (Fragoso-Martínez et al., 2018; González-Gallegos et al. 2018), and nuclear loci
from massive sequencing data (Lara-Cabrera et al. 2021). Using the phylogeny of Salvia subgenus Calosphace as a
tool to target specic metabolites as proposed by Ortiz-Mendoza et al. (2022), we decided explore the phytochem-
istry of key species; since we analyzed a species that was closely related to S. amarissima, which belongs to a taxon
that has shown a wide variety of metabolites, which include neo-clerodanes, 9,10-seco-neo-clerodanes, and amaris-
sanes (Bautista et al. 2016). The major metabolites found in S. urica: amarissinin A (1) and 5,6-dihydroxy-7,3’,4’-
trimethoxyavone (2), also are produced in S. amarissima, supporting their phylogenetic closeness. In addition, the
occurrence of compounds 1 and 2 supports the hypothesis that S. urica has similar pharmacological effects like to its
congener. A previous study analyzed the inhibitory effect of methanolic extracts of plants on the production of vero-
The data are expressed as mean ± S.E.M, n = 6; *P < 0.05 compared to the initial value; aP <0.05 compared
to DM2 control 2h, b P <0.05 compared to DM2 control 4h. SID2: Streptozotocin-induced type 2 diabetic
mice, SUE: Acetone extract of Salvia urica; 1: Amarissinin A; 2: 5,6-dihydroxy-7,3’,4’-trimethoxyavone.
Botanical Sciences / Online First
Botanical Sciences / Online First
7
García-Nava et al. / Botanical Sciences
toxin by the enterohemorrhagic strain of Escherichia coli O157:H7 (VEc) (Sakagami et al. 2001). In that study, the
extract from S. urica showed an inhibitory effect on VEc. Likewise, Calzada et al. (2020) reported that compounds 1
and 2, isolated from S. amarissima, showed antiprotozoal activity against Entamoeba histolytica and Giardia lamblia
with IC50 values ranging from moderate to good activity (62.1-101.1 and 0.05-0.13 μM, respectively), compared with
positive control (0.23-1.22 μM, metronidazole). In the present study, the compounds 1 and 2, but now isolated from
S. urica, showed an antipropulsive effect on charcoal-gum Arabic-induced hyperperistalsis model in rats, providing
new insights to understand underlying ways to stop diarrhea; and provided evidence-based support for the traditional
uses of both plant species. Concerning to antidiabetic effect, previous reports have shown that the antihyperglycemic
activity of a total extract from S. amarissima is associated with an α-glucosidase inhibitory activity. The subsequent
phytochemical analysis of the extract led to the isolation of compound 2, as the more active constituent (IC50 1 800
μM) of the extract. This activity was even higher than those for diterpenes (IC50 > 10 000 μM) also isolated, and close
near to positive control (IC50 100 ± 0.3 μM, acarbose) (Flores-Bocanegra et al. 2017, Solares-Pascacio et al. 2021).
Considering the above, it is plausible that the presence of compound 2 contribute signicantly to the antihyperglyce-
Treatment Hyperperistalsis
(cm)
Antipropulsive
Effect (%)
CE50
(mg/kg)
2 h
AC 75.3 ± 2 - -
AC-GA 91.6 ± 2.7 - -
AC-GA + SUE 50 mg/kg 78 ± 1 83.6 ± 6.6 16.65
25 mg/kg 80.3 ± 1 69.3 ± 6.2
12.5 mg/kg 85.3 ± 0.6 38.7 ± 3.8
AC-GA + 1 1.5 mg/kg 79.3 ± 1.6 75.5 ± 10.4 0.81
0.5 mg/kg 83.6 ± 1.6 48.9 ± 10.1
0.25 mg/kg 86 ± 1.4 34.6 ± 9
0.125 mg/kg 91 ± 0.4 4 ± 2.4
AC-GA + 2 1.5 mg/kg 63.1 ± 1.19 100 0.79
0.5 mg/kg 86.3 ± 0.2 32.6 ± 1.4
0.25 mg/kg 90 ± 0.4 10.2 ± 2.4
0.125 mg/kg 97.3 ± 1.3 0
AC-GA + C 5 mg/kg 39.6 ± 6.5 100 1.003
2.5 mg/kg 77.6 ± 6.1 85.7 ± 5.2
1.25 mg/kg 83 ± 0.4 46.9 ± 2.4
Table 3. Antipropulsive effect Salvia urica and loperamide products on charcoal activated-gum Arabic induced hyperperistalsis
model in rat.
Antipropulsive effect calculated after administration of the treatments. Values expressed as means ± SEM,
n = 6, CE50: half maximal effective concentration. AC: Activated charcoal; GA: Gum Arabic; SUE: Ac-
etone extract of Salvia urica; 1: Amarissinin A; 2: 5,6-dihydroxy-7,3’,4’-trimethoxyavone; C: Loperamide
chloride.
Botanical Sciences / Online First
Botanical Sciences / Online First
Chemical constituents of Salvia urica
8
mic effect of S. urica, through the inhibition of α-glucosidase. In addition, it is worthy to note that was not detected the
presence of oleanolic and ursolic acids, and stigmasterol in the extract of S. urica; which in turn, have been identied
as the bioactive constituents, responsible for the antihyperglycemic properties of Salvia species (Santos et al. 2012,
Bakrim et al. 2022, Ortega et al. 2022). The triterpenoids α- and β-amyrins were also detected in signicant amounts
in the acetone-soluble extract from S. urica by GC-MS (Supplementary material, Table S1). The relative amount (%)
of each triterpenoid in the volatile portion of the extract was: 17.38 ± 8.70 and 25.84 ± 0.98, respectively. Both com-
pounds have been reported as α-glucosidase inhibitors, and are involved in the improving of the insulin plasm levels
and exert a protective effect of β-cells; and prevent the resistance to insulin via AMPK signaling induction (Mabhida
et al. 2018, Dirir et al. 2022, Entezari et al. 2022). Due the above, amyrins are considered as multi-target compounds,
therefore additional studies with the acetone extract of S. urica and their individual constituents are necessary to ex-
plore other action mechanisms that led to understand the antihyperglycemic effect of the plant.
In conclusion, the phytochemical study of an acetone-soluble extract from the aerial parts of S. urica led to the
identication of its major constituents, their quantication through HPLC-PDA, and the evaluation of their antihy-
perglycemic and antiperistaltic effects, providing evidence-based support for the traditional medicinal uses of the
plant. Finally, the pharmacological effect displayed by the diterpenes derived from Salvia amarissima as multidrug-
resistance modulators in cancer cells (Bautista et al. 2016), could prompt exploring these properties in Salvia urica.
Acknowledgments
The authors thank to Dr. Gerardo A. Salazar IB-UNAM, for the photo of Salvia urica kindly provided.
Declaration of competing interests
The authors declare that there is no conict of interest, nancial or personal, in the information, presentation of data
and results of this article.
Supplementary material
Supplemental material for this article can be accessed here https://doi.org/10.17129/botsci.3368
Literature cited
Bakrim S, Benkhaira N, Bourais I, Benali T, Lee LH, Omari NE, Sheikh RA, Goh KW, Ming LC, Bouyahya A.
2022. Health Benets and Pharmacological Properties of Stigmasterol. Antioxidants 11 : 1912. DOI: https://doi.
org/10.3390/antiox11101912
Bautista E, Fragoso-Serrano M, Ortiz-Pastrana N, Toscano RA, Ortega A. 2016. Structural elucidation and evalua-
tion of multidrug-resistance modulatory capability of amarissinins A–C, diterpenes derived from Salvia amaris-
sima. Fitoterapia 114: 1-6. DOI: https://doi.org/10.1016/j.tote.2016.08.007
Bautista E, Lozano-Gamboa S, Fragoso-Serrano M, Rivera-Chávez J, Salazar-Olivo LA. 2022. Jatrophenediol, a
pseudoguaiane sesquiterpenoid from Jatropha dioica rhizomes. Tetrahedron Letters 104: 154040. DOI: https://
doi.org/10.1016/j.tetlet.2022.154040
Calzada F, Arista R, Pérez H. 2010. Effect of plants used in Mexico to treat gastrointestinal disorders on char-
coal-gum acacia-induced hyperperistalsis in rats. Journal of Ethnopharmacology 128: 49-51. DOI: https://doi.
org/10.1016/j.jep.2009.12.022
Calzada F, Bautista E, Barbosa E, Salazar-Olivo LA, Alvidrez-Armendáriz E, Yepez-Mulia L. 2020. Antiprotozoal
activity of secondary metabolites from Salvia circinata. Revista Brasileira de Farmacognosia 30: 593-596. DOI:
https://doi.org/10.1007/s43450-020-00077-7
Botanical Sciences / Online First
Botanical Sciences / Online First
9
García-Nava et al. / Botanical Sciences
Chicago Natural History Museum. 1973. Fieldiana Botany. In: Chicago Natural History Museum 24: 300. USA.
Clebsch B. 2003. The new book of Salvias: Sages for every garden. Portland, USA: Timber Press. ISBN-13: 978-0-
88192-913-3
Dirir AM, Daou M, Yousef AF, Yousef LF. 2022. A review of alpha-glucosidase inhibitors from plants as poten-
tial candidates for the treatment of type-2 diabetes. Phytochemistry Reviews 21: 1049-1079. DOI: https://doi.
org/10.1007/s11101-021-09773-1
Entezari M, Hashemi D, Taheriazam A, Zabolian A, Mohammadi S, Fakhri F, Hashemi M, Hushmandi K, Ashraza-
deh M, Zarrabi A, Nuri-Ertas Y, Mirzaei S, Samarghandian S. 2022. Biomedicine & Pharmacotherapy 146:
112563. DOI: https://doi.org/10.1016/j.biopha.2021.112563
Epling C. 1939 A revision of Salvia subgenus Calosphace. Repertorium Specierum Novarum Regni Vegetabilis.
Dahlem, Berlin: Repertoriums.
Epling C. 1941 Supplementary notes on American Labiatae. II. Bulletin Torrey Botanical Club 68:552-568.
Flores-Bocanegra L, González-Andrade M, Bye R, Linares E, Mata R. 2017. α-Glucosidase Inhibitors from Salvia
circinata. Journal of Natural Products 80: 1584-1593. DOI: https://doi.org/10.1021/acs.jnatprod.7b00155
Fragoso-Martínez I, Martínez-Gordillo M, Salazar GA, Sazatornil F, Jenks AA, García-Peña MR, Barrera-Aveleida
G, Benítez-Vieyra S, Magallón S, Cornejo-Tenorio G, Granados-Mendoza C. 2018. Phylogeny of the Neotropical
sages (Salvia subg. Calosphace; Lamiaceae) and insights into pollinator and area shifts. Plant Systematics and
Evolution 304: 43-55, https://doi.org/10.1007/s00606-017-1445-4
García-Nava X, Fragoso-Serrano M, de Loera D, Cortezano-Arellano O, Calzada F, Bedolla-García BY. 2022. Ama-
risolide H and 15-epi-Amarisolide H, two diterpenoid glucosides from Salvia circinnata. Revista Brasileira de
Farmacognosia 32: 993-999. DOI: https://doi.org/10.1007/s43450-022-00332-z
González-Gallegos JG, Fragoso-Martínez I, González-Adame G, Martínez-Ambriz AE, López-Enríquez IL. 2018.
Salvia ozolotepecensis, S. patriciae and S. sirenis (Lamiaceae), three new species from Miahuatlán district,
Oaxaca, Mexico. Phytotaxa 362: 143-159. DOI: https://doi.org/10.11646/phytotaxa.362.2.2
Harley RM, Atkins S, Budantsev AL, Cantino PD, Conn BJ, Grayer R, Harley MM, de Kok R, Krestovskaja T, Mo-
rales R, Paton AJ, Ryding O, Upson T. 2005. The Families and Genera of Vascular Plants. Taxon, 54: 574. DOI:
https://doi.org/10.2307/25065407
Jenks A, Kim SC. 2013. Medicinal plant complexes of Salvia subgenus Calosphace: An ethnobotanical study of new
world sages. Journal of Ethnopharmacology 146: 214-224. DOI: https://doi.org/10.1016/j.jep.2012.12.035
Lara-Cabrera SI, Perez-Garcia MdlL, Maya-Lastra CA, Montero-Castro JC, Godden GT, Cibrian-Jaramillo A, Fisher
AE, Porter JM. 2021. Phylogenomics of Salvia L. subgenus Calosphace (Lamiaceae). Frontiers in Plant Science
12: 725900. DOI: https://doi.org/10.3389/fpls.2021.725900
Mabhida SE, Dludla PV, Johnson R, Ndlovu M, Louw J, Opoku AR, Mosa RA. 2018. Protective effect of triterpenes
against diabetes-induced β-cell damage: An overview of in vitro and in vivo studies. Pharmacological Research
137: 179-192. DOI: https://doi.org/10.1016/j.phrs.2018.10.004
Martínez-Gordillo M, Bedolla-García B, Cornejo-Tenorio G, Fragoso-Martínez I, García-Peña MdR, González-
Gallegos JG, Lara-Cabrera SI, Zamudio S. 2017. Lamiaceae de México. Botanical Sciences 95:780-806. DOI:
https://doi.org/10.17129/botsci.1871
Ortega A, Pastor-Palacios G, Ortiz-Pastrana N, Ávila-Cabezas E, Toscano RA, Joseph-Nathan P, Morales-Jiménez J,
Bautista E. 2020. Further galphimines from a new population of Galphimia glauca. Phytochemistry 169: 112180.
DOI: https://doi.org/10.1016/j.phytochem.2019.112180
Ortega R, Valdés M, Alarcón-Aguilar FJ, Fortis-Barrera A, Barbosa E, Velázquez C, Calzada F. 2022. Antihypergly-
cemic Effects of Salvia polystachya Cav. and Its Terpenoids: Glucosidase and SGLT1 Inhibitors. Plants 11: 575.
https://doi.org/10.3390/plants11050575
Ortiz-Mendoza N, Aguirre-Hernández E, Fragoso-Martínez I, González-Trujano ME, Basurto-Peña FA, Martínez-
Gordillo MJ. 2022. A review on the ethnopharmacology and phytochemistry of the Neotropical sages (Salvia
Botanical Sciences / Online First
Botanical Sciences / Online First
Chemical constituents of Salvia urica
10
subgenus Calosphace; Lamiaceae) emphasizing Mexican species. Frontiers in Pharmacology 13: 867-892. DOI:
https://doi.org/10.3389/fphar.2022.867892
Padilla-Gómez E. 2007. Estudio ecológico y etnobotánico de la vegetación del Municipio de San Pablo Etla, Oaxa-
ca. Ms Thesis. Instituto Politécnico Nacional. α
Sakagami Y, Murata H, Nakanishi T, Inatomi Y, Watabe K, Iinuma M, Tanaka T, Murata J, Lang FA. 2001. Inhibitory
effect of plant extracts on production of Verotoxin by enterohemorrhagic Escherichia coli O157:H7. Journal of
Health Sciencie 47: 437-477. DOI: https://doi.org/10.1248/jhs.47.473
Santos FA, Frota JT, Arruda BR, de Melo TS, da Silva AA, Brito GAC, Chaves MH, Rao VS. 2012. Antihyperglyce-
mic and hypolipidemic effects of α, β-amyrin, a triterpenoid mixture from Protium heptaphyllum in mice. Lipids
in Health and Disease 11: 98. DOI: https://doi.org/10.1186/1476-511X-11-98
SEMARNAT. [secretaria del Medio Ambiente y Recursos Naturales]. 1999. NOM-062-ZOO-1999: Especicaciones
técnicas para la producción, cuidado y uso de los animales de laboratorio. Ciudad de México: Diario Ocial de la
Federación (miércoles 22 de agosto de 2001).
Sepúlveda-Cuellar L, Duque-Ortiz A, Yáñez-Espinosa L, Calzada F, Bautista E, Pastor-Palacios G, Bedolla García
BY, Flores-Rivera J, Badano EI, Douterlungne D. 2021. Phylogenetic and Chemical Analyses of the Medicinal
Plant Salvia circinnata: an Approach to Understand Metabolomics Differences. Revista Brasileira de Farmacog-
nosia 31: 676-688. DOI: https://doi.org/10.1007/s43450-021-00168-z
Solares-Pascacio JI, Ceballos G, Calzada F, Barbosa E, Velazquez C. 2021. Antihyperglycemic and Lipid Prole
Effects of Salvia amarissima Ortega on Streptozocin-Induced Type 2 Diabetic Mice. Molecules 26: 947. DOI:
https://doi.org/10.3390/molecules26040947
Valdés M, Calzada F, Mendieta-Wejebe JE. 2019. Structure-activity relationship study of acyclic terpenes in blood
glucose levels: potential α-glucosidase and sodium glucose cotransporter (SGLT-1) inhibitors. Molecules 24:
4020. DOI: https://doi.org/10.3390/molecules24224020
Associate editor: Juan Rodrígo Salazar
Author contributions: EB designed the project, provided nancial support, and carried out the phytochemical study. XGN carried out the
phytochemical study. FC and MV conducted the biological assays. OCA and DL acquired and analyzed spectroscopic and spectrometric data.
IFM and MMG collected and identied the plant material studied. All the authors contributed to the writing of the manuscript and approved the
nal version.
Supporting agencies: Consejo Nacional de Ciencia y Tecnologia (CONACYT, project CB-A1-S-7705). X. Garcia (CVU 740455) and E. Bau-
tista are grateful to CONACYT for the Research Fellowships.
Botanical Sciences / Online First
Botanical Sciences / Online First
ResearchGate has not been able to resolve any citations for this publication.
Article
Full-text available
Stigmasterol is an unsaturated phytosterol belonging to the class of tetracyclic triterpenes. It is one of the most common plant sterols, found in a variety of natural sources, including vegetable fats or oils from many plants. Currently, stigmasterol has been examined via in vitro and in vivo assays and molecular docking for its various biological activities on different metabolic disorders. The findings indicate potent pharmacological effects such as anticancer, anti-osteoarthritis, anti-inflammatory, anti-diabetic, immunomodulatory, antiparasitic, antifungal, antibacterial, antioxidant, and neuroprotective properties. Indeed, stigmasterol from plants and algae is a promising molecule in the development of drugs for cancer therapy by triggering intracellular signaling pathways in numerous cancers. It acts on the Akt/mTOR and JAK/STAT pathways in ovarian and gastric cancers. In addition, stigmasterol markedly disrupted angiogenesis in human cholangiocarcinoma by tumor necrosis factor-α (TNF-α) and vascular endothelial growth factor receptor-2 (VEGFR-2) signaling down-regulation. The association of stigmasterol and sorafenib promoted caspase-3 activity and down-regulated levels of the anti-apoptotic protein Bcl-2 in breast cancer. Antioxidant activities ensuring lipid peroxidation and DNA damage lowering conferred to stigmasterol chemoprotective activities in skin cancer. Reactive oxygen species (ROS) regulation also contributes to the neuroprotective effects of stigmasterol, as well as dopamine depletion and acetylcholinesterase inhibition. The anti-inflammatory properties of phytosterols involve the production of anti-inflammatory cytokines, the decrease in inflammatory mediator release, and the inhibition of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Stigmasterol exerts anti-diabetic effects by reducing fasting glucose, serum insulin levels, and oral glucose tolerance. Other findings showed the antiparasitic activities of this molecule against certain strains of parasites such as Trypanosoma congolense (in vivo) and on promastigotes and amastigotes of the Leishmania major (in vitro). Some stigmasterol-rich plants were able to inhibit Candida albicans, virusei, and tropicalis at low doses. Accordingly, this review outlines key insights into the pharmacological abilities of stigmasterol and the specific mechanisms of action underlying some of these effects. Additionally, further investigation regarding pharmacodynamics, pharmacokinetics, and toxicology is recommended.
Article
Full-text available
Salvia is the most diverse genus within the mint family (Lamiaceae), many of its species are well-known due to their medicinal and culinary uses. Most of the ethnopharmacological and phytochemical studies on Salvia are centred on species from the European and Asian clades. However, studies about the most diverse clade, the Neotropical sages (Salvia subgenus Calosphace; 587 spp.), are relatively scarce. This review aims to compile the information on the traditional medicinal uses, pharmacological and phytochemistry properties of the Neotropical sages. To do so, we carried out a comprehensive review of the articles available in different online databases published from the past to 2022 (i.e., PubMed, Scopus, and Web of Science, among others) and summarized the information in tables. To uncover phylogenetic patterns in the distribution of four different groups of metabolites (mono-, sesqui-, di-, and triterpenes), we generated presence-absence matrices and plotted the tip states over a dated phylogeny of Salvia. We found several studies involving Mexican species of Salvia, but only a few about taxa from other diversity centres. The main traditional uses of the Mexican species of Calosphace are medicinal and ceremonial. In traditional medicine 56 species are used to treat diseases from 17 categories according to the WHO, plus cultural-bound syndromes. Pharmacological studies reveal a wide range of biological properties (e.g., antinociceptive, anti-inflammatory, anxiolytic, cytotoxic, and antidiabetic, etc.) found in extracts and isolated compounds of 38 Neotropical sages. From extracts of these species, at least 109 compounds have been isolated, identified and evaluated pharmacologically; 73 of these compounds are clerodanes, 21 abietanes, six flavonoids, five sesquiterpenoids, and four triterpenoids. The most characteristic metabolites found in the Neotropical sages are the diterpenes, particularly clerodanes (e.g., Amarisolide A, Tilifodiolide), that are found almost exclusively in this group. The Neotropical sages are a promising resource in the production of herbal medication, but studies that corroborate the properties that have been attributed to them in traditional medicine are scarce. Research of these metabolites guided by the phylogenies is recommended, since closely related species tend to share the presence of similar compounds and thus similar medicinal properties.
Article
Full-text available
The antihyperglycemic activity of ethanolic extract from Salvia polystachya (EESpS) and its products was evaluated using in vivo, ex vivo and in silico assays; additionally, an acute toxicity assay was evaluated. EESpS was classified as a nontoxic class 5 drug. EESpS, ethyl acetate fraction (EtOAcFr), secondary-6-fraction (SeFr6), ursolic acid (UA), and oleanolic acid (OA) reduced the hyperglycemia in DM2 mice. α-glucosidase inhibition was evaluated with oral sucrose and starch tolerance tests (OSuTT and OStTT), an intestinal sucrose hydrolysis (ISH) assay and molecular docking studies using acarbose as control. SGLT1 inhibition was evaluated with oral glucose and galactose tolerance tests (OGTT and OGaTT), an intestinal glucose absorption (IGA) assay and molecular docking studies using canagliflozin as the control. During the carbohydrate tolerance tests, all the treatments reduced the postprandial peak, similar to the control drugs. During the ISH, IC50 values of 739.9 and 726.3 µM for UA and OA, respectively, were calculated. During the IGA, IC50 values of 966.6 and 849.3 for UA, OA respectively, were calculated. Finally, during the molecular docking studies, UA and OA showed ∆G values of −6.41 and −5.48 kcal/mol−1, respectively, on α-glucosidase enzymes. During SGLT1, UA and OA showed ∆G values of −10.55 and −9.65, respectively.
Article
Full-text available
Diabetes mellitus (DM) is considered as a main challenge in both developing and developed countries, as lifestyle has changed and its management seems to be vital. Type I and type II diabetes are the main kinds and they result in hyperglycemia in patients and related complications. The gene expression alteration can lead to development of DM and related complications. The AMP-activated protein kinase (AMPK) is an energy sensor with aberrant expression in various diseases including cancer, cardiovascular diseases and DM. The present review focuses on understanding AMPK role in DM. Inducing AMPK signaling promotes glucose in DM that is of importance for ameliorating hyperglycemia. Further investigation reveals the role of AMPK signaling in enhancing insulin sensitivity for treatment of diabetic patients. Furthermore, AMPK upregulation inhibits stress and cell death in β cells that is of importance for preventing type I diabetes development. The clinical studies on diabetic patients have shown the role of AMPK signaling in improving diabetic complications such as brain disorders. Furthermore, AMPK can improve neuropathy, nephropathy, liver diseases and reproductive alterations occurring during DM. For exerting such protective impacts, AMPK signaling interacts with other molecular pathways such as PGC-1α, PI3K/Akt, NOX4 and NF-κB among others. Therefore, providing therapeutics based on AMPK targeting can be beneficial for amelioration of DM.
Article
Full-text available
The evolutionary relationships of Salvia have been difficult to estimate. In this study, we used the Next Generation Sequencing method Hyb-Seq to evaluate relationships among 90 Lamiaceae samples, including representatives of Mentheae, Ocimeae, Salvia subgenera Audibertia, Leonia, Salvia, and 69 species of subgenus Calosphace, representing 32 of Epling's sections. A bait set was designed in MarkerMiner using available transcriptome data to enrich 119 variable nuclear loci. Nuclear and chloroplast loci were assembled with hybphylomaker (HPM), followed by coalescent approach analyses for nuclear data (ASTRAL, BEAST) and a concatenated Maximum Likelihood analysis of chloroplast loci. The HPM assembly had an average of 1,314,368 mapped reads for the sample and 527 putative exons. Phylogenetic inferences resolved strongly supported relationships for the deep-level nodes, agreeing with previous hypotheses which assumed that subgenus Audibertia is sister to subgenus Calosphace. Within subgenus Calosphace, we recovered eight monophyletic sections sensu Epling, Cardinalis, Hastatae, Incarnatae, and Uricae in all the analyses (nDNA and cpDNA), Biflorae, Lavanduloideae, and Sigmoideae in nuclear analyses (ASTRAL, BEAST) and Curtiflorae in ASTRAL trees. Network analysis supports deep node relationships, some of the main clades, and recovers reticulation within the core Calosphace. The chloroplast phylogeny resolved deep nodes and four monophyletic Calosphace sections. Placement of S. axillaris is distinct in nuclear evidence and chloroplast, as sister to the rest of the S. subg. Calosphace in chloroplast and a clade with "Hastatae clade" sister to the rest of the subgenus in nuclear evidence. We also tested the monophyly of S. hispanica, S. polystachia, S. purpurea, and S. tiliifolia, including two samples of each, and found that S. hispanica and S. purpurea are monophyletic. Our baits can be used in future studies of Lamiaceae phylogeny to estimate relationships between genera and among species. In this study, we presented a Hyb-Seq phylogeny for complex, recently diverged Salvia, which could be implemented in other Lamiaceae.
Article
Full-text available
The evolutionary relationships of Salvia have been difficult to estimate. In this study, we used the Next Generation Sequencing method Hyb-Seq to evaluate relationships among 90 Lamiaceae samples, including representatives of Mentheae, Ocimeae, Salvia subgenera Audibertia, Leonia, Salvia, and 69 species of subgenus Calosphace, representing 32 of Epling's sections. A bait set was designed in MarkerMiner using available transcriptome data to enrich 119 variable nuclear loci. Nuclear and chloroplast loci were assembled with hybphylomaker (HPM), followed by coalescent approach analyses for nuclear data (ASTRAL, BEAST) and a concatenated Maximum Likelihood analysis of chloroplast loci. The HPM assembly had an average of 1,314,368 mapped reads for the sample and 527 putative exons. Phylogenetic inferences resolved strongly supported relationships for the deep-level nodes, agreeing with previous hypotheses which assumed that subgenus Audibertia is sister to subgenus Calosphace. Within subgenus Calosphace, we recovered eight monophyletic sections sensu Epling, Cardinalis, Hastatae, Incarnatae, and Uricae in all the analyses (nDNA and cpDNA), Biflorae, Lavanduloideae, and Sigmoideae in nuclear analyses (ASTRAL, BEAST) and Curtiflorae in ASTRAL trees. Network analysis supports deep node relationships, some of the main clades, and recovers reticulation within the core Calosphace. The chloroplast phylogeny resolved deep nodes and four monophyletic Calosphace sections. Placement of S. axillaris is distinct in nuclear evidence and chloroplast, as sister to the rest of the S. subg. Calosphace in chloroplast and a clade with “Hastatae clade” sister to the rest of the subgenus in nuclear evidence. We also tested the monophyly of S. hispanica, S. polystachia, S. purpurea, and S. tiliifolia, including two samples of each, and found that S. hispanica and S. purpurea are monophyletic. Our baits can be used in future studies of Lamiaceae phylogeny to estimate relationships between genera and among species. In this study, we presented a Hyb-Seq phylogeny for complex, recently diverged Salvia, which could be implemented in other Lamiaceae.
Article
The HPLC–PDA profiling of an acetone-soluble extract of the leaves of Salvia circinnata Cav. (Syn. Salvia amarissima Ortega), Lamiaceae, collected at Patzcuaro, Michoacan, Mexico, indicated this population corresponds to the chemotype “amarisolide A.” The phytochemical study of the extract led to the isolation of two new diterpenoid glucosides, amarisolide H and 15-epi-amarisolide H, which were obtained as an epimeric mixture, together with the known compounds amarisolides A, D, G, 16-epi-amarisolide G, and 5,6-dihydroxy-7,3′,4′-trimethoxyflavone. The structures of compounds amarisolide H and 15-epi-amarisolide H were determined by the analyses of their NMR and HRMS data. The cytotoxicity and anti-multidrug resistance (MDR) effects of compounds amarisolide H, 15-epi-amarisolide H, amarisolide A, amarisolide G, and 16-epi-amarisolide G in MCF-7 cancer cells sensible and resistant to vinblastine were essayed.
Article
Jatrophenediol (1), a new pseudoguaiane sesquiterpenoid, was isolated from the roots of Jatropha dioica, together with the known diterpenoids citlalitrione (2) and jatropolones A and B (3 and 4). The structure of 1 was established based on the analysis of its spectroscopic (1D/2D NMR) and spectrometric (HRESIMS) data. The cytotoxic effect of all isolates was assayed against a panel of six cancer cell lines (HeLa, MCF-7, PC-3, HCT-15, HCT-116, and RAW 264.7). Unfortunately, no cytotoxicity was detected for compounds 1-4. Additionally, the reversion of multi-drug-resistance (MDR) to vinblastine was explored in three phenotypes of the MCF-7 line. Interestingly, the mixture of compounds 3/4 in three different ratios at 25 μg/mL increased the sensitivity to vinblastine by 7-10-fold.
Article
Salvia ozolotepecensis, S. patriciae and S. sirenis (Lamiaceae) from Oaxaca, Mexico, are described and illustrated as three new species. The first one is characterized by its glandular-capitate pubescence in stems, leaves and inflorescences, mostly persistent floral bracts, entire and 5-veined upper calyx lip, red corolla, epapillate inside, included stamens, and connective ornate with opposed retrorse and antrorse teeth bordering an incision. It is most similar to S. perlonga and S. praestans to which it is compared. The second can be distinguished in having ovate to ovate-deltoid leaf blades, entire, acute and 7-veined upper calyx lips, red corollas, internally ornate near the base with two slender linguiform papillae, upper corolla lip shorter than the lower one, exserted stamens and glabrous style. This new taxon is morphologically similar to the species belonging to Salvia sections Cardinales, Charantia, Incarnatae, Flexuosa, Fulgentes, Iodophyllae and Pedicellata; although, it cannot be unambiguously assigned to one of these. In contrast, S. sirenis clearly belongs to S. sect. Scorodonia; amongst the species therein, the new species can be differentiated by the concolorous leaves, rounded to slightly oblique at base, 6-8-flowered floral nodes, longer floral bracts, pedicels, calyces and corollas, and 5 or 7-veined upper calyx lip. Also, it represents an addition to Mexican Salvia with white corollas. The molecular markers ITS, trnL-trnF and trnH-psbA were sequenced for two of the three new species, and their phylogenetic position is discussed.