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Aqueous Bark Extract of Ceiba speciosa (A. St.-Hill) Ravenna Protects against Glucose Toxicity in Caenorhabditis elegans

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Oxidative Medicine and Cellular Longevity
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Plants are widely used in folk medicine because of their pharmacological properties. Ceiba speciosa, popularly known as paineira-rosa or tree-of-wool, is a species found in the Northwest of Rio Grande do Sul, being native of the upper Uruguay River, Brazil. The tea obtained from the stem bark is employed in folk medicine to reduce cholesterol, triacylglycerides, and glucose levels. However, there are no studies in the literature proving its efficacy or the safety of its use. For this study, we used Caenorhabditis elegans as an animal model considering its advantages for risk assessment and pharmacological screenings. For the toxicological tests, C. elegans N2 (wild type) was treated with the aqueous extract of the stem bark of C. speciosa (ECE) at the first larval stage (L1) at concentrations of 5, 25, 50, and 250 μg/mL. To evaluate biological activities, we challenged the extract for oxidative stress resistance in the presence of paraquat (0.5 mM), H2O2 (1 mM), and against glucose-induced toxicity. Our results demonstrated that ECE did not alter survival rate, pharyngeal pumping, and reproduction of the nematodes. The extract was not able to protect the nematodes against the toxicity induced by prooxidants. Notably, ECE protected against glucotoxicity by increasing worms' life span and by reducing glucose levels. On the other hand, ECE treatment did not reduce lipid accumulation induced by exogenous glucose feeding, as observed in worms which lipid droplets were tagged with GFP. Based on our results, we believe that the extract is indeed promising for further studies focusing on carbohydrates metabolism; however, it needs to be carefully evaluated since the extract does not seem to modulate lipid accumulation.
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Research Article
Aqueous Bark Extract of Ceiba speciosa (A. St.-Hill) Ravenna
Protects against Glucose Toxicity in Caenorhabditis elegans
Fabrine Bianchin dos Santos,
1
Caroline Brandão Quines ,
1
Luiz Eduardo Ben Pilissão,
1
Ana Helena de Castro Dal Forno,
1
Cristiane Freitas Rodrigues,
1
Cristiane Casagrande Denardin ,
1
Fabiane Moreira Farias,
2
and Daiana Silva Ávila
1
1
Laboratório de Pesquisa em Bioquímica e Toxicologia em Caenorhabditis elegans, Programa de Pós-Graduação em Bioquímica,
Universidade Federal do Pampa, Uruguaiana, Brazil BR 472-Km 592-Caixa Postal 118, CEP 97500-970
2
Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Pampa, Uruguaiana, Brazil BR 472-Km 592-
Caixa Postal 118, CEP 97500-970
Correspondence should be addressed to Daiana Silva Ávila; avilads1@gmail.com
Received 29 April 2020; Revised 1 July 2020; Accepted 25 July 2020; Published 8 October 2020
Academic Editor: Márcio Carocho
Copyright © 2020 Fabrine Bianchin dos Santos et al. This is an open access article distributed under the Creative Commons
Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work
is properly cited.
Plants are widely used in folk medicine because of their pharmacological properties. Ceiba speciosa, popularly known as paineira-
rosa or tree-of-wool, is a species found in the Northwest of Rio Grande do Sul, being native of the upper Uruguay River, Brazil. The
tea obtained from the stem bark is employed in folk medicine to reduce cholesterol, triacylglycerides, and glucose levels. However,
there are no studies in the literature proving its ecacy or the safety of its use. For this study, we used Caenorhabditis elegans as an
animal model considering its advantages for risk assessment and pharmacological screenings. For the toxicological tests, C. elegans
N2 (wild type) was treated with the aqueous extract of the stem bark of C. speciosa (ECE) at the rst larval stage (L1) at
concentrations of 5, 25, 50, and 250 μg/mL. To evaluate biological activities, we challenged the extract for oxidative stress
resistance in the presence of paraquat (0.5 mM), H
2
O
2
(1 mM), and against glucose-induced toxicity. Our results demonstrated
that ECE did not alter survival rate, pharyngeal pumping, and reproduction of the nematodes. The extract was not able to
protect the nematodes against the toxicity induced by prooxidants. Notably, ECE protected against glucotoxicity by increasing
wormslife span and by reducing glucose levels. On the other hand, ECE treatment did not reduce lipid accumulation induced
by exogenous glucose feeding, as observed in worms which lipid droplets were tagged with GFP. Based on our results, we believe
that the extract is indeed promising for further studies focusing on carbohydrates metabolism; however, it needs to be carefully
evaluated since the extract does not seem to modulate lipid accumulation.
1. Introduction
Glucose toxicity refers to the biological eects of its excessive
levels in cells and tissues due to metabolic alterations and
production of advanced glycation end products (AGEs) [1].
AGES interact with lipids, proteins, and other molecules
causing cell damage by inducing oxidative stress and cell
death [2]. With the growing interest in nding natural anti-
oxidants and drugs with benecial action on glucose metab-
olism, many extracts from dierent plants and their
dierent parts have been tested [3, 4]. However, the indis-
criminate use of natural products has led to cases of intoxica-
tion [5]. Therefore, a safety assessment of natural products
should be performed previously to medicinal use [4].
The scientic literature describes some medicinal proper-
ties for several species of the genus Ceiba. A study conducted
by Leal et al. [6] reported a considerable variety of secondary
compounds in the bark extract and antimicrobial activity
against strains of Staphylococcus aureus which may be related
to the presence of phytochemicals such as tannins and phe-
nols. In addition, Ceiba pentandra (L.) Gaertn, commonly
known as silk cotton tree, has been used by practitioners of
traditional medicine in Northern and Eastern Nigeria for dia-
betes control.
Hindawi
Oxidative Medicine and Cellular Longevity
Volume 2020, Article ID 1321354, 9 pages
https://doi.org/10.1155/2020/1321354
In this same pharmacological context, Ceiba speciosa (A.
St.-Hil.) has been used by the southern Brazilian population
[7]. The species is found in the Northwestern Rio Grande
do Sul, Brazil, being native in the Upper Uruguay vegetation
[8]. Regarding its use in folk medicine, there are records for
the use of barks and owers for the treatment of heart dis-
ease, hypertension, and diabetes [9]. With the increasing
interest in the bioprospection of natural products, especially
of vegetal material originating from the Pampa Biome, the
validation and application of alternative models for the eval-
uation of these natural products becomes relevant.
Among the animal models that have been used to inves-
tigate the toxicology and pharmacology of medicinal plants is
the nematode Caenorhabditis elegans. This worm has a num-
ber of characteristics that make it not only relevant but also
quite powerful as a model for this type of research [10]. C. ele-
gans is easy and cheap to maintain in the laboratory with an
Escherichia coli diet, has a short reproductive cycle (3 days)
and a large number of descendants (~200), which allow the
large-scale production of animals within a short period of
time. The transparent body allows a clear observation of all
cells in these animals. Notably, C. elegans shows a strong con-
servation in molecular and cellular pathways with respect to
mammals and humans [11], including those signaling related
to glucose and lipid metabolism [12, 13].
Notably, this nematode does not have specialized storage
tissues such as adipocytes, and the energetic excess is stored
in lipid droplets in enterocytes, hypodermis, and gonads
[14, 15]. On the other hand, the absence of adipocytes
removes central components in the control of energetic
metabolism in mammals, which may be advantageous by
facilitating the interpretation of other conserved factors and
mechanisms involved in energy homeostasis [16, 17]. Thus,
this study is aimed at evaluating the safety of the aqueous
bark extract from Ceiba speciosa (ECE) as well as at verifying
the glucose-reducing levels action attributed to this plant
using C. elegans as an animal model.
2. Materials and Methods
2.1. Extract Preparation. The stem bark of C. speciosa was
collected in the municipality of Santo Antônio das Missões,
Rio Grande do Sul, Brazil, on April 2012. The plant material
was identied by a botanist, and an exsiccata or botanical
voucher was deposited in the Zoobotanic Herbarium
Augusto Ruschi at the University of Passo Fundo, under
the code RSPF 12637. C. speciosa barks were ground, and
the aqueous extract was prepared by decoction in distilled
water according to Malheiros et al. [18]. Since we have used
the exact same batch of lyophilized extract, the phytochemi-
cal composition has been already reported [18]. To expose
worms, the lyophilized material was diluted in water to
obtain a stock solution at 10 mg/mL that was subsequently
used for further dilutions.
2.2. Caenorhabditis elegans Maintenance. The C. elegans
strains used in this study were N2 (wild type) and VS29
(hjSi56 [vha-6p::3xFLAG::TEV::GFP::dgat-2::let-858 3
UTR]) [19] which were obtained from Caenorhabditis
Genetics Center (CGC) and maintained in NGM (nematode
growth medium) seeded with E. coli OP50 at 20
°
C [20]. First
larval stage worms were obtained by a synchronization pro-
cess, which consists of exposing pregnant worms to the lysis
solution (0.45 N NaOH, 2% HOCl p/v) to separate the eggs
from the worms. After 14 h, the eggs hatched and released
the L1 larvae, used for exposures. All experiments were per-
formed at 22
°
C in a controlled humidied environment.
2.3. Treatment Protocol and LC
50
Determination. A total of
2,000 L1 nematodes were exposed to dierent concentrations
of C. speciosa bark extract (ECE-5 μg/mL, 25 μg/mL,
50 μg/mL, and 250 μg/mL) for 30 min at 22
°
C, in a homoge-
nizer. After treatment, the nematodes were washed with
85 mM NaCl solution for 3 times in order to terminate the
exposure, and then, nematodes were plated on NGM seeded
with E. coli OP50. Twenty-four hours after exposure, the
number of surviving nematodes was counted, and a survival
curve was drawn [21]. The experiments were performed in
duplicates. The results were expressed as % of control.
2.4. Brood Size. The nematodes were treated as described
above and were maintained on NGM/E. coli OP50 plates
until they reached larval stage L4. To evaluate progeny size,
one nematode from each ECE treatment was transferred to
a new plate containing NGM medium, and the total number
of progeny was counted during the whole reproductive
period [22]. The experiments were performed in triplicates
and independently repeated three times. The results were
expressed as % of control.
2.5. Pharyngeal Pumping. This assay is used to verify the die-
tary intake of the nematodes. 24 h after the treatment
described previously, 5 nematodes from each experiment
were transferred to new plates without bacteria. The pharyn-
geal pumps were counted for 1 min at the stereomicroscope
[23]. The experiments were repeated in three independent
assays. The results were expressed as % of control.
2.6. Oxidative Stress-Resistance Assays. Nematodes were pre-
treated at the L1 stage as described previously. After the last
wash, worms were posttreated with the prooxidants paraquat
(0.5 mM) or hydrogen peroxide (1 mM) for 30 min. Subse-
quently, nematodes were washed four more times and then
transferred to NGM/E. coli OP50 plates. The number of sur-
viving nematodes on each plate 24 h postexposure was scored
to determine stress resistance [10]. The experiments were
performed in duplicates and repeated at least 3 times. The
results were expressed as % of control.
2.7. Glucose Toxicity. In the pilot test, we have used 2 glucose
concentrations (2 and 4%) to verify whether they would
reduce wormslife span. Plates containing NGM medium
were inoculated with E. coli OP50 which were UV-
inactivated. Thereafter, glucose solution was added on top
of the dried E.coli. Based on pilot results, we have chosen glu-
cose 4% for the next assays. Worms were pretreated with dif-
ferent concentrations of th e extract (ECE 5 μg/mL or
250 μg/mL) for 30 min, washed to remove the treatments,
and then transferred to NGM/UV-inactivated E.coli plates
2 Oxidative Medicine and Cellular Longevity
containing glucose 4% or vehicle. Then, 20 live worms at the
same developmental stage were collected at the late L4 stage
and transferred daily to new plates NGM UV-inactivated
E.coli + glucose 4% or vehicle in order to prevent contamina-
tion or progeny. Live animals were scored until all animals
died. The experiments were performed in triplicates, and
three independent experiments were conducted. The results
were expressed as % of control.
2.8. Glucose Levels. The glucose content was quantied by
glucose oxidase method. Worms (N2) were treated as
described in Section 2.7 and kept in glucose 4% until they
reached the L4 stage. After these 48 h, worms were washed
owith M9 buer from the plates and centrifuged at
2500 rpm for 2 min. Worms were washed until removing all
existing bacteria. Then, worms were frozen and thawed 3
times, sonicated with lysis buer, and centrifuged. The super-
natant (50 μL) was transferred to a 96-well plate, and glucose
levels were measured using a Labtest glucose colorimetric
assay kit (Labtest Diagnostica S.A., Minas Gerais, Brasil).
The optical density of each of the incubated samples
(10 min at 37
°
C) was measured at 505 nm. For normalization
of the data, protein was measured using Bradford colorimet-
ric method and expressed as mg glucose/μg protein. The
experiments were performed in duplicates and normalized
as % of control.
2.9. Glucose-Induced Lipid Accumulation. N2 and VS29 nem-
atodes were treated as described in Section 2.7, and worms
were kept on NGM/UV-inactivated E.coli OP50 plates
containing or not glucose 4% until reaching the larval
stage L4. After these 48 hours of glucose exposure, worms
were transferred to slides containing levamisole, and
images were acquired in a uorescent microscope to
observe the eect of glucose exposure on lipid accumula-
tion. VS29 has a transgene which DGAT2 (acyl CoA:dia-
cylglycerol O-acyltransferase 2) is fused to GFP. This
enzyme transfers acyl groups and is localized in the lipid
particles; then, lipid droplets can be observed. N2 was
used as control to verify lipofuscin or autouorescence
production caused by glucose treatment.
2.10. Statistical Analysis. Statistical analysis was carried out
by one-way analysis of variance (ANOVA), followed by post
hoc Tukey test when the overall p<0:05. For life span assay,
a repeated measure two-way analysis of variance (ANOVA)
followed by post hoc Tukey test was applied.
3. Results
3.1. Toxicity Evaluation of C. speciosa Aqueous Bark Extract.
Figure 1(a) shows that acute exposure to dierent concentra-
tions of ECE did not alter C. elegans survival rate (up to
250 μg/mL), thus suggesting that no toxic eects occurred
under the experimental conditions tested (acute). Based on
that, we decided to use the concentrations of 5 μg/mL,
25 μg/mL, 50 μg/mL, and 250 μg/mL for further assays. In
addition, we did not nd any toxic eect of C. speciosa bark
extract on wormsreproduction, as brood size did not change
(Figure 1(b)). In order to verify whether ECE would aect
food ingestion per se, we analyzed pharyngeal pumping,
which was not changed as well (Figure 1(c)).
3.2. Stress Resistance. To verify a possible stress resistance
conferred by ECE, we exposed worms to dierent prooxi-
dants: paraquat (a pesticide, Figure 2(a)) or hydrogen perox-
ide (H
2
O
2
, Figure 2(b)). Our results demonstrate that ECE
did not protect against the damage induced by these prooxi-
dants at any of the tested concentrations.
3.3. ECE Treatment Protected against Glucose Toxicity. In
order to verify the protection of ECE treatment against glu-
cose toxicity, we analyzed life span. Post hoc analysis of the
data depicted in Figure 3 has revealed that 4% glucose
induced a reduction in wormslife span from day 5 until
day 14. The treatment with ECE at both concentrations (5
and 250 μg/mL) was eective against the glucose-induced
reduction in life span starting on day 6 until day 12. Two-
way ANOVA demonstrated a signicant interaction between
treatment ×days (Fð54,162Þ=4:418;p<0:001), thus indicat-
ing a protection conferred by ECE throughout the life span.
In addition, glucose 4% administration signicantly
increased glucose levels, and the treatment with ECE was
eective by reducing this elevation (Figure 4(b)). Notably,
ECE per se treatment did not alter glucose levels
(Figure 4(a)).
Furthermore, we took advantage of a GFP-tagged strain
and observed lipid accumulation induced by glucose vs.
ECE pretreatment. In wild-type (N2) worms, no increase or
decrease in uorescence intensity caused by lipofuscin pro-
duction was evidenced, even in those worms exposed to
high glucose levels (Figures 5(a)5(d)). On the other hand,
in VS29 mutants, a stronger uorescence around the gut
following glucose 4% exposure was detected, therefore
indicating an increase in lipid droplets accumulation
(Figure 6(b)). However, ECE treatment did not reduce lipid
accumulation at any of the concentrations tested since no
signicant reduction in GFP uorescence was evidenced
(Figures 6(c) and 6(d)).
4. Discussion
In the present work, the safety and pharmacological activity
of C. speciosa aqueous bark extract were studied, since little
scientic knowledge about its eects are described in the lit-
erature. The consumption of this plant occurs in the North-
western region of the State of Rio Grande do Sul, Brazil,
which consumes it as a tea due to its alleged benecial eects
in humans [9]. Our work veried that ECE is safe and pro-
tected against glucose-induced toxicity by reducing its levels
and the associated consequences in C. elegans life span.
The antioxidant potential of the extract has been previ-
ously demonstrated in vitro through the DPPH radical scav-
enging capacity and demonstrated a good scavenging
potential [18, 24]. The phytochemical analysis of the extract
used in this study was published by Malheiros and cols and
revealed that avonoids (quercetin, rutin, and kaempferol)
and phenolic acids (gallic, chlorogenic, ellagic, and caeic)
are present [18, 25].
3Oxidative Medicine and Cellular Longevity
Survival (% of control)
Control Paraquat 5 25 50 250
0
50
100
150
Paraquat+Ceiba
speciosa
(𝜇g/mL)
⁎⁎ ⁎⁎ ⁎⁎
(a)
Survival (% of control)
Control H2O25 25 50 250
0
50
100
150
200
H2O2+Ceiba speciosa (𝜇g/mL)
⁎⁎⁎ ⁎⁎⁎ ⁎⁎⁎ ⁎⁎⁎
⁎⁎⁎
(b)
Figure 2: Eect of dierent concentrations of ECE treatment in oxidative stress resistance. (a) Response to oxidative stress in paraquat
posttreated nematodes (0.5 mM) and pretreated with ECE. Values are expressed as mean ± S:E:M:of six experiments. (b) Response to
oxidative stress in post-H
2
O
2
treated nematodes (1 mM) and pretreated with the ECE. Values are expressed as mean ± S:E:M:of three
experiments. Data were analyzed by using one-way ANOVA following by the Tukey post hoc test. indicates p<0:05;∗∗ indicates
p<0:01;∗∗∗ indicates p<0:001 when compared to the control group.
Control 5 25 50 250
0
50
100
150
Ceiba speciosa (𝜇g/mL)
Survival (% of control)
(a)
0
100
200
300
Ceiba speciosa (𝜇g/mL)
Brood size (% of control)
Control 5 25 50 250
(b)
Ceiba speciosa (𝜇g/mL)
Control
52550250
0
50
100
150
Pharyngeal pumping
(% of control)
(c)
Figure 1: Toxicological evaluation of dierent concentrations of C. speciosa aqueous extract in (a) nematodes survival, (b) brood size, and (c)
pharyngeal pumping. Values are expressed as mean ± S:E:M:of four experiments. Data were analyzed by using one-way ANOVA following
by the Tukey test.
4 Oxidative Medicine and Cellular Longevity
⁎⁎⁎
⁎⁎⁎
##
⁎⁎⁎
⁎⁎⁎
⁎⁎
⁎⁎
###
⁎⁎⁎
⁎⁎
⁎⁎⁎
###
###
###
⁎⁎⁎
⁎⁎⁎
### ###
⁎⁎⁎
#
⁎⁎
#
5101520
0
20
40
60
80
100
Days
Longevity (% of control)
### ###
Control
Glucose
Glucose+5 𝜇g/mL
Glucose+250 𝜇g/mL
Figure 3: Eect of dierent concentration of ECE on glucose-induced toxicity in wormslife span. Values are expressed as mean ± S:E:M:of
four experiments. Data were analyzed by using a repeated measures two-way ANOVA followed by the Tukey posttest. indicates p<0:05;
∗∗ indicates p<0:01;∗∗∗ indicates p<0:001 when compared to the control group.
#
indicates p<0:05;
##
indicates p<0:01;
###
indicates
p<0:001 when compared to the glucose group.
Ceiba speciosa (𝜇g/mL)
N2 5 25 50 250
0
50
100
150
Glucose/
𝜇
g of protein
(% of control)
(a)
Glucose/
𝜇
g of protein
(% of control)
N2 Glucose 5 25 50 250
0
50
100
150
200
250
Glucose+Ceiba speciosa
(𝜇g/mL)
###
#
#
(b)
Figure 4: Glucose levels in (a) following ECE treatment per se and (b) worms treated with ECE and exposed to glucose 4%. Values are
expressed as mean ± S:E:M:of four experiments. Data were analyzed by using one-way ANOVA following by the Tukey post hoc test.
indicates p<0:05 when compared to the control group.
#
indicates p<0:05;
###
indicates p<0:001 when compared to the glucose group.
5Oxidative Medicine and Cellular Longevity
In vivo, however, the extract did not show the same anti-
oxidant performance. We challenged ECE against two pow-
erful prooxidants, paraquat and H
2
O
2
, which have
signicantly increased nematodes mortality. Paraquat is an
herbicide widely used in agriculture, but its biochemical
mechanism responsible for its toxicity is not fully
(a) (b)
(c) (d)
Figure 5: Representative images of N2 worms following glucose-induced lipid-accumulation and ECE treatment to observe autouorescence:
(a) untreated worms; (b) worms treated with glucose 4%; (c) worms treated with ECE 5 μg/mL+ 4% glucose; (d) worms treated with ECE
250 μg/mL+ 4% glucose.
(a) (b)
(c) (d)
Figure 6: Representative images of VS29 worms following glucose-induced lipid-accumulation and ECE treated worms: (a) untreated worms;
(b) worms exposed to glucose 4%; (c) worms treated with ECE 5 μg/mL+ 4% glucose; (d) worms treated with ECE 250 μg/mL+ 4% glucose.
6 Oxidative Medicine and Cellular Longevity
understood. It is known that paraquat is a superoxide rad-
ical generator (O2·), which can produce other reactive spe-
cies such as hydrogen peroxide (H
2
O
2
) and hydroxyl
radical (·OH), which are unstable and react quickly with
fatty acids, causing lesions in the membranes, proteins,
and DNA. H
2
O
2
is one of the most versatile oxidants in
existence, superior to chlorine, chlorine dioxide, and
potassium permanganate; through catalysis, H
2
O
2
can be
converted to hydroxyl radical (·OH). It can easily pass
through cell membranes [26, 27]. Our results demon-
strated that despite the antioxidant action in vitro, the
treatment with ECE was not able to reduce the toxicity
induced by these prooxidants. In vivo, molecules can be
transformed, sometimes losing activities presented in some
in vitro assays, particularly those without tissue.
C. elegans contains many key metabolic-related com-
ponents such as oxidative stress and insulin signaling
pathways, making it a useful platform to improve our
understanding of complex diseases such as metabolic syn-
drome [13]. Glucose is the main energy source and key
regulator of animal metabolism; however, a chronic abun-
dance of glucose will have deleterious eects on cellular
and tissue functions [28]. Carbohydrate excess activates
several lipogenic enzymes contributing to deregulation in
lipid metabolism and insulin resistance development [29].
Furthermore, high levels of glucose induce cell injury of
mammalian hepatocytes and pancreatic cells through
molecular mechanisms of endoplasmic reticulum stress,
oxidative stress, and mitochondrial impairment [29, 30].
Moreover, chronic hyperglycemia can deteriorate cognitive
function and impair memory [31].
C. elegans is a suitable model organism to study glucose
toxicity, once high glucose conditions impair their life span
by increasing ROS formation and AGEs modication of
mitochondrial proteins in a DAF-2 independent manner
[32]. Furthermore, glucose-enriched diet signicantly
decreases wormslife span by inhibiting the translocation of
DAF-16 and HSF-1 transcription factors that are involved
in antioxidant response [33]. On the other hand, glucose
restriction extends C. elegans life span by increasing the
expression of proaging and antioxidant genes [34].
We have found promising results when we challenged
ECE-treated worms against high glucose levels. The extract
was able to protect from the reduced longevity and to dimin-
ish whole body glucose levels induced by high glucose expo-
sure. A similar eect has been observed in the alloxan-
induced diabetic rats treated with B. ceiba bark methanolic
extract [35]. Furthermore, the alcoholic extract of C. pentan-
dra, a species taxonomically related to C. speciosa which is
commonly used in North Africa, has been used to control
hyperglycemia in diabetic patients [36]. Notably, its alcoholic
extract has been tested in rats, and it has been proven that
besides safe, decreases plasma glucose levels following oral
administration [37].
In relation to mechanism of action, the treatment with C.
pentandra decreased glucose levels in diabetic rats through
enhanced glucose utilization and regulation in insulin levels
[38, 39]. Similar to C. pentandra, we believe that the treat-
ment with ECE could enhance glucose utilization and stimu-
late another form of energy storage, like glycogen or
trehalose. Notably, C. elegans can synthesize saturated and
unsaturated fatty acids and is capable of storing energy as
lipids (triglycerides, phospholipids, sphingolipids, etc.). Up
to 35% of C. elegans dry body mass is lipids, being triacylgly-
ceride fat deposits the major energy storage molecules,
depending on diet and life stage [16]. To analyze that, we
have used a transgenic strain which acyl CoA:diacylglycerol
O-acyltransferase 2 is fused to GFP. This enzyme transfers
the acyl groups and is localized in the lipid particles; then,
lipid droplets can be observed in vivo. As expected, our
results have shown that high glucose administration
increased lipid accumulation in worms. However, ECE treat-
ment failed to reduce this storage, therefore indicating that
glucose levels reduced by the extract may have been con-
verted to lipids.
Finally, we observed that acute exposure at dierent
concentrations of ECE did not cause toxicological eects
on worms, as analyzed by survival rate, brood size, and
pharyngeal pumping. These ndings conrm that worms
were ingesting the extract and feeding normally, indicat-
ing that the use of these concentrations is safe, consider-
ing the experimental conditions tested. Previous in vitro
data have also demonstrated that this same extract did
not reduce leukocytes viability and did not cause geno-
toxicity [18]. C. speciosa is a plant widely used by south-
ern Brazilians; therefore, the safety of its use by this
population is germane. Thus, for the rst time, it was
demonstrated that the aqueous bark extract of C. speciosa
has low toxicity and has potential against glucose toxic-
ity, thus suggesting that ECE may be useful against
hyperglycemia in mammals. Therefore, additional studies
will be carried out by our group with the purpose of
expanding the hypoglycemic eects of C. speciosa and
its mechanisms.
Data Availability
All authors declare the availability of the use of data from this
manuscript by the journal Antioxidants and Prooxidants:
Eects on Health and Aging 2020.
Conflicts of Interest
The authors declare that they have no conicts of interest.
Acknowledgments
The authors are thankful to PROPPI/Universidade Federal
do Pampa (UNIPAMPA) for providing nancial assistance.
DSA is recipient of CNPq researcher scholarship and sup-
ported by FAPERGS/PqG # 18/2551-0000434-0. AHCDF
and CFR were recipients of scholarships from Coordenação
de Aperfeiçoamento de Pessoal de Nível SuperiorBrasil
(CAPES)Finance Code 001. Some strains were provided
by the CGC, which is funded by NIH Oce of Research
Infrastructure Programs (P40 OD010440).
7Oxidative Medicine and Cellular Longevity
Supplementary Materials
The supplementary material includes the graphical abstract
of the manuscript. (Supplementary Materials)
References
[1] X. Luo, J. Wu, S. Jing, and L. J. Yan, Hyperglycemic stress and
carbon stress in diabetic glucotoxicity,Aging and Disease,
vol. 7, no. 1, pp. 90110, 2016.
[2] A. Pinkas and M. Aschner, Advanced glycation end-products
and their receptors: related pathologies, recent therapeutic
strategies, and a potential model for future neurodegeneration
studies,Chemical Research in Toxicology, vol. 29, no. 5,
pp. 707714, 2016.
[3] S. Surveswaran, Y. Z. Cai, J. Xing, H. Corke, and M. Sun,
Antioxidant properties and principal phenolic phytochemi-
cals of Indian medicinal plants from Asclepiadoideae and
Periplocoideae,Natural Product Research, vol. 24, no. 3,
pp. 206221, 2010.
[4] A. M. Pisoschi, A. Pop, C. Cimpeanu, and G. Predoi, Antiox-
idant capacity determination in plants and plant-derived
products: a review,Oxidative Medicine and Cellular Longev-
ity, vol. 2016, Article ID 9130976, 36 pages, 2016.
[5] K. Boubaker, M. Ounissi, N. Brahmi et al., Acute renal failure
by ingestion of Euphorbia paralias,Saudi Journal of Kidney
Diseases and Transplantation, vol. 24, no. 3, pp. 571575,
2013.
[6] I. C. R. Leal, K. R. N. dos Santos, I. I. Júnior et al., Ceanothane
and lupane type triterpenes from Zizyphus joazeiroan anti-
staphylococcal evaluation,Planta Medica, vol. 76, no. 1,
pp. 4752, 2010.
[7] E. Beleski-Carneiro, J. A. Sugui, and F. Reicher, Structural and
biological features of a hydrogel from seed coats of Chorisia
speciosa,Phytochemistry, vol. 61, no. 2, pp. 157163, 2002.
[8] L. Cappelatti and J. L. Schmitt, Caracterização da ora
arbórea de um fragmento urbano de oresta estacional semi-
decidual no Rio Grande do Sul, Brasil, Pesquisas,Botânica,
vol. 60, pp. 341354, 2009.
[9] R. F. P. de Lucena, V. T. do Nascimento, E. de Lima Araújo,
and U. P. de Albuquerque, Local uses of native plants in
an area of Caatinga vegetation (Pernambuco, NE Brazil),
Ethnobotany Research and Applications, vol. 6, pp. 003
014, 2008.
[10] J. H. An and T. K. Blackwell, SKN-1 links C. elegans mesen-
dodermal specication to a conserved oxidative stress
response,Genes & Development, vol. 17, no. 15, pp. 1882
1893, 2003.
[11] C. Weinhouse, L. Truong, J. N. Meyer, and P. Allard, Caenor-
habditis elegans as an emerging model system in environmen-
tal epigenetics,Environmental and Molecular Mutagenesis,
vol. 59, no. 7, pp. 560575, 2018.
[12] J. Zheng and F. L. Greenway, Caenorhabditis elegans as a
model for obesity research,International Journal of Obesity,
vol. 36, no. 2, pp. 186194, 2012.
[13] J. L. Watts and M. Ristow, Lipid and carbohydrate metabo-
lism in Caenorhabditis elegans,Genetics, vol. 207, no. 2,
pp. 413446, 2017.
[14] K. Ashra,Obesity and the regulation of fat metabolism,
WormBook: The Online Review of C. Elegans Biology, pp. 1
20, 2007.
[15] K. Palikaras, M. Mari, B. Petanidou, A. Pasparaki,
G. Filippidis, and N. Tavernarakis, Ectopic fat deposition
contributes to age-associated pathology in Caenorhabditis ele-
gans,Journal of Lipid Research, vol. 58, no. 1, pp. 7280, 2017.
[16] K. Ashra, F. Y. Chang, J. L. Watts et al., Genome-wide RNAi
analysis of Caenorhabditis elegans fat regulatory genes,
Nature, vol. 421, no. 6920, pp. 268272, 2003.
[17] S. Hashmi, Y. Wang, R. S. Parhar et al., AC. elegans model to
study human metabolic regulation,Nutrition & Metabolism,
vol. 10, no. 1, p. 31, 2013.
[18] C. K. C. Malheiros, J. S. B. Silva, T. C. Hofmann et al., Prelim-
inary in vitro assessment of the potential toxicity and antioxi-
dant activity of Ceiba speciosa (A. St.-Hill) Ravenna
(Paineira),Brazilian Journal of Pharmaceutical Sciences,
vol. 53, no. 2, 2017.
[19] N. Xu, S. O. Zhang, R. A. Cole et al., The FATP1-DGAT2
complex facilitates lipid droplet expansion at the ER-lipid
droplet interface,The Journal of Cell Biology, vol. 198, no. 5,
pp. 895911, 2012.
[20] S. Brenner, The genetics of Caenorhabditis elegans,Genetics,
vol. 77, no. 1, pp. 7194, 1974.
[21] D. Avila, K. Helmcke, and M. Aschner, The Caenorhabiditis
elegans model as a reliable tool in neurotoxicology,Human
& Experimental Toxicology, vol. 31, no. 3, pp. 236243, 2012.
[22] Y. Guo, Y. Yang, and D. Wang, Induction of reproductive
decits in nematode Caenorhabditis elegans exposed to metals
at dierent developmental stages,Reproductive Toxicology,
vol. 28, no. 1, pp. 9095, 2009.
[23] D. Raizen, B.-m. Song, N. Trojanowski, and Y.-J. You,
Methods for measuring pharyngeal behaviors,2005.
[24] M. N. Yesmin, S. N. Uddin, S. Mubassara, and M. A. Akond,
Antioxidant and antibacterial activities of Calotropis procera
Linn,American-Eurasian Journal of Agricultural & Environ-
mental Sciences, vol. 4, pp. 550553, 2008.
[25] C. K. C. Malheiros, Avaliação Preliminar In Vitro do Potencial
Antioxidante e da Toxicidade de Ceiba Speciosa (a. St.-hill)
Ravenna (paineira), 2014.
[26] J. Azadmanesh and G. E. O. Borgstahl, A review of the cata-
lytic mechanism of human manganese superoxide dismutase,
Antioxidants, vol. 7, no. 2, p. 25, 2018.
[27] H. Younus, Therapeutic potentials of superoxide dismutase,
International Journal of Health Sciences, vol. 12, no. 3, pp. 88
93, 2018.
[28] M. Soto-Garcia, M. Rosales-Castro, G. N. Escalona-Cardoso,
and N. Paniagua-Castro, Evaluation of hypoglycemic and
genotoxic eect of polyphenolic bark extract fromQuercus
sideroxyla,Evidence-Based Complementary and Alternative
Medicine, vol. 2016, Article ID 4032618, 7 pages, 2016.
[29] M. Mota, B. A. Banini, S. C. Cazanave, and A. J. Sanyal,
Molecular mechanisms of lipotoxicity and glucotoxicity in
nonalcoholic fatty liver disease,Metabolism, vol. 65, no. 8,
pp. 10491061, 2016.
[30] C. Liu, Y. Hao, F. Yin, Y. Zhang, and J. Liu, Geniposide pro-
tects pancreatic βcells from high glucose-mediated injury by
activation of AMP-activated protein kinase,Cell Biology
International, vol. 41, no. 5, pp. 544554, 2017.
[31] M. J. Overman, N. Pendleton, T. W. O'Neill et al., Glycemia
but not the metabolic syndrome is associated with cognitive
decline: ndings from the European Male Ageing Study,
The American Journal of Geriatric Psychiatry, vol. 25, no. 6,
pp. 662671, 2017.
8 Oxidative Medicine and Cellular Longevity
[32] A. Schlotterer, G. Kukudov, F. Bozorgmehr et al., C. elegans as
model for the study of high glucose- mediated life span reduc-
tion,Diabetes, vol. 58, no. 11, pp. 24502456, 2009.
[33] S. J. Lee, C. T. Murphy, and C. Kenyon, Glucose shortens the
life span of C. elegans by downregulating DAF-16/FOXO
activity and aquaporin gene expression,Cell Metabolism,
vol. 10, no. 5, pp. 379391, 2009.
[34] T. J. Schulz, K. Zarse, A. Voigt, N. Urban, M. Birringer, and
M. Ristow, Glucose restriction extends Caenorhabditis ele-
gans life span by inducing mitochondrial respiration and
increasing oxidative stress,Cell Metabolism, vol. 6, no. 4,
pp. 280293, 2007.
[35] R. Zahan, L. Nahar, M. Haque, M. L. Nesa, and Z. Alam, Anti-
oxidant and antidiabetic activities of Alangium salvifolium and
Bombax ceiba,Dhaka University Journal of Pharmaceutical
Sciences, vol. 12, no. 2, pp. 159163, 2015.
[36] A. Majeed, A. A. El-Sayed, T. Khoja, R. Alshamsan, C. Millett,
and S. Rawaf, Diabetes in the Middle-East and North Africa:
an update,Diabetes Research and Clinical Practice, vol. 103,
no. 2, pp. 218222, 2014.
[37] C. K. Foe, E. P. Nguelefack-Mbuyo, N. Tsabang, A. Kamanyi,
and T. B. Nguelefack, Hypoglycemic properties of the aque-
ous extract from the stem bark of Ceiba pentandra in
dexamethasone-induced insulin resistant rats,Evidence-
Based Complementary and Alternative Medicine, vol. 2018,
Article ID 4234981, 11 pages, 2018.
[38] R. Satyaprakash, M. Rajesh, M. Bhanumathy et al., Hypogly-
cemic and antihyperglycemic eect of Ceiba pentandra l.
gaertn in normal and streptozotocin-induced diabetic rats,
Ghana Medical Journal, vol. 47, no. 3, pp. 121127, 2013.
[39] O. Ladeji, I. Omekarah, and M. Solomon, Hypoglycemic
properties of aqueous bark extract of Ceiba pentandra in
streptozotocin-induced diabetic rats,Journal of Ethnophar-
macology, vol. 84, no. 2-3, pp. 139142, 2003.
9Oxidative Medicine and Cellular Longevity
... In this work, the head thrashes were measured in the absence and presence of E. coli. In absence of E. coli, the C. does not alter the survival of C. elegans adult and the brood size (Santos et al., 2020). These authors show that C. elegans adults were ingesting the extract and feeding normally (Santos et al., 2020). ...
... In absence of E. coli, the C. does not alter the survival of C. elegans adult and the brood size (Santos et al., 2020). These authors show that C. elegans adults were ingesting the extract and feeding normally (Santos et al., 2020). C. elegans adults treated with S. schottiana extract remained active, even in absence of E. coli. ...
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Helminthiasis is considered a serious illness in tropical countries. The study of activity of natural products has proven to be an excellent alternative to parasite control. This work evaluated the anthelmintic activity of the crude extract of Stachytarpheta schottiana on viability, movements, reproduction behaviors and morphology of different stages of Caenorhabditis elegans. The extract did not decrease L1 hatching, and the survival of L1-L2 and adults in all tested concentrations. And it did not alter the movements of C. elegans adults and the egg-laying activity and brood size. Chemotaxis behavioral assays showed that C. elegans was not attracted or repelled by the extract. The morphological analysis of different forms showed several alterations as cuticular damages that promoted the detachment from the nematode body. The eggs were degraded or not present in adult bodies treated. The results obtained with the crude extract suggest that several secondary metabolites isolated from S. schottiana have an anthelmintic effect and for this confirmation, future analyses will be necessary.
... The JB1E and JB2E extracts increased the sensitivity of C. elegans to paraquat, which is known as a pro-oxidant due to its ability to generate reactive oxygen species [71], and this effect was most pronounced in the JB1E extract. Paraquat is used in agriculture as an herbicide, and the mechanism of its toxic effect is the induction of reactive oxygen species [72]. The increased sensitivity of C. elegans to paraquat after pre-exposure to the ethanolic extract of juniper fruits suggests that the mechanism of its nematocidal activity is the induction of oxidative stress. ...
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Citation: Belov, T.; Terenzhev, D.; Bushmeleva, K.N.; Davydova, L.; Burkin, K.; Fitsev, I.; Gatiyatullina, A.; Egorova, A.; Nikitin, E. Comparative Analysis of Chemical Profile and Biological Activity of Juniperus communis L. Berry Extracts. Plants 2023, 12, 3401. https://doi. Abstract: Researchers are looking for the most effective ways to extract the bioactive substances of Juniperus communis L. berries, which are capable of displaying the greatest range of biological activity, namely antimicrobial potential "against phytopathogens", antioxidant activity and nematocidal activity. This study provides detailed information on the chemical activity, group composition and biological activity of the extracts of juniper berries of 1-and 2-year maturity (JB1 and JB2), which were obtained by using different solvents (pentane, chloroform, acetone, methanol and 70% ethanol) under various extraction conditions (maceration and ultrasound-assisted maceration (US)). Seventy percent ethanol and acetone extracts of juniper berries were analyzed via gas chromatography-mass spectrometry, and they contained monoterpenes, sesquiterpenes, polysaccharides, steroids, fatty acid esters and bicyclic monoterpenes. The antimicrobial activity was higher in the berries of 1-year maturity, while the acetone extract obtained via ultrasound-assisted maceration was the most bioactive in relation to the phytopathogens. Depending on the extraction method and the choice of solvent, the antioxidant activity with the use of US decreased by 1.5-1.9 times compared to the extracts obtained via maceration. An analysis of the nematocidal activity showed that the sensitivity to the action of extracts in Caenorhabditis elegans was significantly higher than in Caenorhabditis briggsae, particularly for the acetone extract obtained from the juniper berries of 1-year maturity.
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Hymenaea genus has been used in folk medicine in Brazil, but few studies investigated its toxicity profile. Thus, the aim of this study was to determine toxicological parameters of Hymenaea courbaril stem bark hydroalcoholic extract by utilizing three cell lines including murine macrophages (RAW 264.7), mouse fibroblast cells (L929) and human lung fibroblast (MRC-5), as well as Salmonella/microsome assay, and in vivo Caenorhabditis elegans model. The predominant detected phytoconstituents in the extract were coumarins, flavonoids, phenolics, tannins and saponins and by HPLC analysis, astilbin (AST) was found to be the main component. The DPPH assay demonstrated that H. courbaril hydroalcoholic extract exhibited potent antioxidant activity, with an IC50 of 3.12 μg/ml. The extract at concentrations of 400 and 800 μg/ml decreased cell viability 48 hr after treatment in L929 and MRC-5 cell lines. In the Raw 264.7 strain, just the highest concentration (800 μg/ml) lowered cell viability within 48 hr following exposure. The concentration of 100 μg/ml did not markedly affect cell viability in the trypan blue assay. In the alkaline comet assay the extract was found to be non-genotoxic. In the Ames test, the extract exhibited low mutagenic potential without metabolic activation, since only the highest concentrations produced an effect. H. courbaril extract only affected the survival of C. elegans at concentrations of 800 and 1600 μl/ml. These findings demonstrate that H. courbaril extract appears to exert low toxicity as evidenced in vitro and mutagenicity assays; however, the biological relevance of the response of C. elegans survival to safety assessments needs further studies.
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b>Introduction : Neotropical genus Ceiba Mill. is known for having tall trees, trunks with a robust part and fruits of economic interest. In Brazil, there are eleven native species, from those five are endemic. It is used in folk medicine to treat diabetes, inflammation, pain and diarrhea; however, most of the species have no scientific validation for such activities. This review aims to compile information about the genus Ceiba in Brazil, to update and allow an integrated understanding of its medicinal uses, chemical composition, and biological activities. Methodology : Ceiba species reported in Flora e Funga do Brasil were used for this literature review, using ScienceDirect, PubMed, and Google Scholar as articles databases. Results : In traditional use, the most cited species was C. pentandra (25 citations), the most used part was stem/bark (30 citations), preparation method was decoction (19 citations), and main administration route was oral ingestion (12 reports) for digestive system, skin and subcutaneous tissues diseases treatment. Genus chemical composition is wide, presenting metabolites such as proteins, sugars, fatty acids, tannins, flavonoids, and alkaloids. Bioactivities as antidiabetic, antioxidant, antitumor and antimicrobial were observed, especially for C. pentandra . Conclusion : Some species of Ceiba have extensive scientific literature, presenting several isolated compounds and bioactivities. On the other hand, some species, especially those that are endemics only in Brazil, do not have studies that evaluate their biological properties. Such knowledge is essential to confirm the medicinal potential cited in ethnobotanical reports, and the possibility of new compounds of biotechnological interest
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In this work, the extraction procedure of a natural pigment from the flower of Ceiba speciosa (A. St.-Hil.) was optimized by response surface methodology. It is the first time that the extraction of the flower pigment of C. speciosa (FPCS) has been reported, along with an evaluation of its stability and biological activity under various conditions, and an exploration of its potential use as a food additive and in medicine. Specifically, the effects of ethanol concentration, solid–liquid ratio, temperature and time on the extraction rate of FPCS were determined using a Box–Behnken design. The optimum extraction conditions for FPCS were 75% ethanol with a solid–liquid ratio of 1:75 mg/mL) at 66 °C for 39 min. The purification of FPCS using different macroporous resins showed that D101 performed best when the initial mass concentration of the injection solution was 1.50 mg/mL, resulting in a three-fold increase in color value. The yield of dry flowers was 9.75% of fresh petals and the FPCS extraction efficiency was 43.2%. The effects of light, solubility, pH, temperature, sweeteners, edible acids, redox agents, preservatives and metal ions on FPCS were also investigated. Furthermore, the characteristics of FPCS were determined by spectrophotometry at a specific wavelength using the Lambert–Beer law to correlate the mass of FPCS with its absorbance value. An acute toxicological test performed according to Horne’s method showed that FPCS is a non-toxic extract and thus may be used as a food additive or in other ingestible forms. Finally, western blotting showed that FPCS prevents lipopolysaccharide-induced hippocampal oxidative stress in mice. The study suggests that FPCS may function as an antioxidant with applications in the food, cosmetics and polymer industries.
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Parts of Ceiba pentandra are wildly used in Africa to treat diabetes and previous works have demonstrated their in vivo antidiabetic effects on type 1 diabetes models. In addition, it has been recently shown that the decoction and the methanol extract from the stem bark of C. pentandra potentiate in vitro, the peripheral glucose consumption by the liver and skeletal muscle slices. But nothing is known about its effect on type II diabetes, especially on insulin resistance condition. We investigated herein the antihyperglycemic, insulin-sensitizing potential, and cardioprotective effects of the dried decoction from the stem bark of Ceiba pentandra (DCP) in dexamethasone-induced insulin resistant rats. DCP phytochemical analysis using LC-MS showed the presence of many compounds, including 8-formyl-7-hydroxy-5-isopropyl-2-methoxy-3-methyl-1,4-naphthaquinone, 2,4,6-trimethoxyphenol, and vavain. Wistar rats were given intramuscularly ( i.m. ) dexamethasone (1 mg/kg/day) alone or concomitantly with oral doses of DCP (75 or 150 mg/kg/day) or metformin (40 mg/kg/day) for 9 days. Parameters such as body weight, glycemia, oral glucose tolerance, plasma triglycerides and cholesterol, blood pressure, and heart rate were evaluated. Moreover, cardiac, hepatic and aortic antioxidants (reduced glutathione, catalase, and superoxide dismutase), malondialdehyde level, and nitric oxide content were determined. DCP decreased glycemia by up to 34% and corrected the impairment of glucose tolerance induced by dexamethasone but has no significant effect on blood pressure and heart rate. DCP reduced the total plasma cholesterol and triglycerides as compared to animals treated only with dexamethasone. DCP also increased catalase, glutathione, and NO levels impaired by dexamethasone, without any effect on SOD and malondialdehyde. In conclusion, the decoction of the stem bark of Ceiba pentandra has insulin sensitive effects as demonstrated by the improvement of glucose tolerance, oxidative status, and plasma lipid profile. This extract may therefore be a good candidate for the treatment of type II diabetes.
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The roundworm Caenorhabitis elegans has been an established model organism for the study of genetics and developmental biology, including studies of transcriptional regulation, since the 1970s. This model organism has continued to be used as a classical model system as the field of transcriptional regulation has expanded to include scientific advances in epigenetics and chromatin biology. In the last several decades, C. elegans has emerged as a powerful model for environmental toxicology, particularly for the study of chemical genotoxicity. Here, we outline the utility and applicability of C. elegans as a powerful model organism for mechanistic studies of environmental influences on the epigenome. Our goal in this article is to inform the field of environmental epigenetics of the strengths and limitations of the well‐established C. elegans model organism as an emerging model for medium‐throughput, in vivo exploration of the role of exogenous chemical stimuli in transcriptional regulation, developmental epigenetic reprogramming, and epigenetic memory and inheritance. As the field of environmental epigenetics matures, and research begins to map mechanisms underlying observed associations, new toolkits and model systems, particularly manipulable, scalable in vivo systems that accurately model human transcriptional regulatory circuits, will provide an essential experimental bridge between in vitro biochemical experiments and mammalian model systems. Environ. Mol. Mutagen., 2018. © 2018 Wiley Periodicals, Inc.
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Lipid and carbohydrate metabolism are highly conserved processes that affect nearly all aspects of organismal biology. Caenorhabditis elegans eat bacteria, which consist of lipids, carbohydrates, and proteins that are broken down during digestion into fatty acids, simple sugars, and amino acid precursors. With these nutrients, C. elegans synthesizes a wide range of metabolites that are required for development and behavior. In this review, we outline lipid and carbohydrate structures as well as biosynthesis and breakdown pathways that have been characterized in C. elegans We bring attention to functional studies using mutant strains that reveal physiological roles for specific lipids and carbohydrates during development, aging, and adaptation to changing environmental conditions.
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Our previous works indicated that geniposide could regulate glucose-stimulated insulin secretion (GSIS), and improved chronic high glucose-induced dysfunctions in pancreatic β cells, but the molecular mechanisms remain largely unknown. In the present study, we investigated the role of 5 -AMP-activated protein kinase (AMPK) in high glucose induced cell injury and explored the associated molecular mechanisms in rat INS-1 pancreatic β cells. Data suggested that geniposide obviously prevented the cell damage induced by high (25 mM) glucose in INS-1 cells, which increased the protein levels of cell apoptosis-associated enzymes, including heme oxygenase-1 (HO-1) and Bcl-2, but apparently attenuated the protein level of Bax, an apoptotic protein. In addition, Compound C, an AMPK inhibitor, remarkably inhibited the effects of geniposide on the protein levels of HO-1, Bcl-2 and Bax, but AICAR, an AMPK activator, potentiated the role of geniposide on the protein levels of HO-1, Bcl-2 and Bax. More importantly, geniposide directly prevented the cleavage of caspase-3 induced by high glucose, and this effect was also evidently prohibited by the pre-incubation of compound C in high glucose-treated INS-1 cells. Furthermore, using the method of RNA interfere, we further proved that treatment with AMPK siRNA attenuated the effects of geniposide on the apoptosis-associated proteins and cell viability. All these data suggest that AMPK plays a crucial role on geniposide antagonizing high glucose-induced pancreatic β cells injury.
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Objective: Previous research has indicated that components of the metabolic syndrome (MetS), such as hyperglycemia and hypertension, are negatively associated with cognition. However, evidence that MetS itself is related to cognitive performance has been inconsistent. This longitudinal study investigates whether MetS or its components affect cognitive decline in aging men and whether any interaction with inflammation exists. Methods: Over a mean of 4.4 years (SD ± 0.3), men aged 40-79 years from the multicenter European Male Ageing Study were recruited. Cognitive functioning was assessed using the Rey-Osterrieth Complex Figure (ROCF), the Camden Topographical Recognition Memory (CTRM) task, and the Digit Symbol Substitution Test (DSST). High-sensitivity C-reactive protein (hs-CRP) levels were measured using a chemiluminescent immunometric assay. Results: Overall, 1,913 participants contributed data to the ROCF analyses and 1,965 subjects contributed to the CTRM and DSST analyses. In multiple regression models the presence of baseline MetS was not associated with cognitive decline over time (p > 0.05). However, logistic ordinal regressions indicated that high glucose levels were related to a greater risk of decline on the ROCF Copy (β = -0.42, p < 0.05) and the DSST (β = -0.39, p < 0.001). There was neither a main effect of hs-CRP levels nor an interaction effect of hs-CRP and MetS at baseline on cognitive decline. Conclusion: No evidence was found for a relationship between MetS or inflammation and cognitive decline in this sample of aging men. However, glycemia was negatively associated with visuoconstructional abilities and processing speed.