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Effect of boldo (Peumus boldus Molina) infusion on lipoperoxidation induced by cisplatin in mice liver


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Peumus boldus Molina (Monimiaceae), commonly referred to as 'boldo', is used in traditional Chilean medicine to treat hepatic and gastrointestinal diseases. Its leaves are rich in antioxidant compounds, principally alkaloids and flavonoids. This study evaluates the protective effect of a complete boldo leaf infusion on lipoperoxidation (MDA determination at 532 nm) induced by cisplatin in mice liver. To determine if the observed effect can be explained by the action of boldine or catechin, each compound was studied separately. The mice were divided into 8 groups (n = 6): (I) not treated; (II) treated with cisplatin 6 mg/Kg b.w.; (III) treated with boldo leaf infusion 5%; (IV) pretreated with boldo leaf infusion 5% and treated with cisplatin 6 mg/Kg b.w.; (V) treated with boldine 50 mg/Kg b.w.; (VI) pretreated with boldine 50 mg/Kg b.w. and treated with cisplatin 6 mg/kg.b.w.; (VII) treated with catechin; and (VIII) pretreated with catechin 50 mg/Kg b.w. and treated with cisplatin 6 mg/Kg b.w. As expected, the treatment with cisplatin significantly increased (p < 0.01) lipoperoxidation in comparison with the non-treated group. Pretreatment with boldo leaf infusion significantly diminished (p < 0.05) the lipoperoxidation induced by cisplatin with respect to the animals not pretreated with the infusion. The pretreatments with boldine and catechin significantly diminished (p < 0.05) the lipoperoxidation induced by cisplatin with respect to the group treated only with cisplatin. The results suggest that the boldo infusion is acting as a protector with respect to the oxidative hepatic damage caused by cisplatin, and that this protective ability would be due to the presence in the infusion of the natural antioxidants boldine and principally catechin. These findings suggest the potential use of the infusion as a chemoprotector.
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Copyright © 2009 John Wiley & Sons, Ltd. Phytother. Res. 23, 1024–1027 (2009)
DOI: 10.1002/ptr
Copyright © 2009 John Wiley & Sons, Ltd.
Phytother. Res. 23, 1024–1027 (2009)
Published online 14 January 2009 in Wiley InterScience
( DOI: 10.1002/ptr.2746
Effect of boldo (Peumus boldus Molina)
infusion on lipoperoxidation induced by
cisplatin in mice liver
J. Fernández, P. Lagos, P. Rivera and E. Zamorano-Ponce*
Universidad del Bío-Bío, Casilla 447, Chillán, Chile
Peumus boldus Molina (Monimiaceae), commonly referred to as ‘boldo’, is used in traditional Chilean medi-
cine to treat hepatic and gastrointestinal diseases. Its leaves are rich in antioxidant compounds, principally
alkaloids and flavonoids. This study evaluates the protective effect of a complete boldo leaf infusion on
lipoperoxidation (MDA determination at 532 nm) induced by cisplatin in mice liver. To determine if the
observed effect can be explained by the action of boldine or catechin, each compound was studied separately.
The mice were divided into 8 groups (n ==
= 6): (I) not treated; (II) treated with cisplatin 6 mg/Kg b.w.; (III)
treated with boldo leaf infusion 5%; (IV) pretreated with boldo leaf infusion 5% and treated with cisplatin
6mg/Kg b.w.; (V) treated with boldine 50 mg/Kg b.w.; (VI) pretreated with boldine 50 mg/Kg b.w. and treated
with cisplatin 6 mg/kg.b.w.; (VII) treated with catechin; and (VIII) pretreated with catechin 50 mg/Kg b.w. and
treated with cisplatin 6 mg/Kg b.w. As expected, the treatment with cisplatin significantly increased (p <<
< 0.01)
lipoperoxidation in comparison with the non-treated group. Pretreatment with boldo leaf infusion significantly
diminished (p <<
< 0.05) the lipoperoxidation induced by cisplatin with respect to the animals not pretreated with
the infusion. The pretreatments with boldine and catechin significantly diminished (p <<
< 0.05) the lipoperoxidation
induced by cisplatin with respect to the group treated only with cisplatin. The results suggest that the boldo
infusion is acting as a protector with respect to the oxidative hepatic damage caused by cisplatin, and that
this protective ability would be due to the presence in the infusion of the natural antioxidants boldine
and principally catechin. These findings suggest the potential use of the infusion as a chemoprotector.
Copyright © 2009 John Wiley & Sons, Ltd.
Keywords: Peumus boldus Molina, Cisplatin; lipoperoxidation; antioxidants; boldine; catechin.
Even though cisplatin (cis-diaminodicloroplatin) is one
of the most active cytotoxic agents used for decades to
treat diverse solid cancerous tumors, evidence indicates
that treatment with this drug produces several collateral
effects, derived from oxidative stress that is induced at
both the renal and hepatic level (Pratibha et al., 2006).
A large number of natural products and dietary com-
ponents have been evaluated for their capacity to act
as antioxidants (Dragland et al., 2003), including plant
extracts (Cai et al., 2004; Zamorano-Ponce et al., 2004,
2006). Since many of these plant extracts have long
been used in traditional medicine, there is an emerging
scientific interest to better understand the mechanisms
that are involved with the different biological effects
(Valerio and Gonzales, 2005).
Boldo (Peumus boldus Molina), a tree of the Moni-
miaceae family that is endemic to Chile, has been known
for its medicinal properties since it was used by diverse
indigenous groups, including the Mapuche ethnia who
lived in Chile prior to the arrival of the Spanish in
the fifteenth century (Ferrer, 1904). Actually, boldo is
widely used in Chilean folk medicine and is recognized
as a medicinal herb in pharmacopoeia. Its use is
recommended as a regulator of the hepatic function,
colagogue, antispasmodic, digestive stimulant, and
nervous sedative. It is most commonly taken as a leaf
infusion that is drunk after eating (Muñoz et al., 2001).
Boldo leaves are rich in alkaloids, flavonoids, and
essential oils (O’Brien et al., 2006).
The present study evaluates the oxidative damage
induced by cis-DDP in Mus musculus, Balb-C mouse
hepatic tissue and investigates the possible protective
effect provided by the complete infusion of boldo leaves
as well as each of the natural antioxidants present in
boldo leaves, boldine and catechin, separately, deter-
mining the lipoperoxidation (MDA measurement at
532 nm) as an oxidative stress cellular marker (Hwang
and Kim, 2007).
Animals. Males Mus musculus, Balb-C strain, 68 weeks
old, weighing approximately 25 g were supplied by the
Animal House of the Faculty of Science of University
of Bío-Bío. Animal use and care in all experiments
comply with Chilean ethical laws on animal manipula-
tion. Mice were housed in plastic cages at 22 ± 1°C,
60 ± 10% humidity and 12 h light/12 h dark cycle during
Received 15 May 2008
Revised 21 October 2008
Accepted 21 October 2008
* Correspondence to: E. Zamorano-Ponce, Laboratorio de Genética
Toxicológica GENETOX, Departamento de Ciencias Básicas, Facultad
de Ciencias, Universidad del Bío-Bío, Casilla 447, Chillán, Chile.
Copyright © 2009 John Wiley & Sons, Ltd. Phytother. Res. 23, 1024–1027 (2009)
DOI: 10.1002/ptr
Table 1. Protocols and treatment
Treatment Groups (n = 6) Dose
Untreated 1 –
Cis-DDP 2 6 mg/Kg b.w.a
Boldo infusion 3 5%/30 daysb
Infusion/cis-DDP 4 5%, 30 daysb/6 mg/Kg b.w.a
Boldine 5 50 mg/Kg b.w. /30 daysb
Boldine/cis-DDP 6 50 mg/Kg b.w. 30 daysb/6 mg/kg b.w.a
Catechin 7 50 mg/Kg b.w. /30 daysb
Catechin/cis-DDP 8 50 mg/Kg b.w. 30 daysb/6 mg/Kg b.w.a
a Intraperitoneal, 72 hours before sacrifice.
b Oral administration.
the acclimatization period to laboratory conditions
and throughout the entire experimental period. Water
was available ad libitum and the animals were fed a
conventional laboratory diet (pellet Kimber®). Mice
were separated in eight experimental groups with six
animals each, as shown in the schedule presented in
Table 1.
Once each treatment was complete, the animals were
sacrificed by cervical dislocation. The liver (0.5 g) was
removed, washed with cold sodium phosphate buffer
(50 mM, pH = 7.0), and homogenized in sodium phos-
phate buffer to give a 10% w/v homogenate. The homo-
genate was centrifuged at 750 × g for 10 min and the
supernatant (1 ml) was used for malondialdehyde
(MDA) determination by thiobarbituric acid reaction
(Ohkawa et al., 1979). The absorbance was measured
at 532 nm (Spectronic® Genesys™ .5 spectrophoto-
meters) and compared with those obtained from MDA
standards (
= 2, 23 × 105M1cm1).
Plant material. Fresh Peumus Boldus Molina leaves
were sampled in March 2006 in the Campus Fernando
May, Universidad del Bío-Bío, Chillán, Chile (S 36°
36 05,1, W 72° 04 38,1) at 120 m above sea level.
The plant material was identified by Dr Patricio
Peñailillo Brito at the Institute of Plant Biology and
Biotechnology, Universidad de Talca, Chile. Voucher
specimen (No. 3332) has been kept at the Herbarium
of Universidad de Talca. Lead samples were dried at
room temperature.
Preparation of infusion. The infusion was made by
pouring 100 ml of boiling water on 5 g of plant mate-
rial. The mixture was left to stand for 30 min, filtered,
and then used. The infusion was freshly prepared for
each experiment.
Chemical and dosing. All chemicals were purchased
from Sigma Chemical Company (St Louis, MO, USA).
Boldine was used at a concentration of 50 mg/Kg b.w.
(Jimenez and Speisky, 2000). To prepare the solution
of boldine, it was first dissolved in ethanol and once
the ethanol was evaporated, the aqueous solution of
Boldine was prepared. Catechin was used at a concen-
tration of 50 mg/Kg b.w. (Isbrucker et al., 2006). The
dose of cis-DDP was 6 mg/Kg b.w.i.p. (Sadzuca, 1991).
The chemical solution dosing volume was determined
based on each animal’s body weight, considering a
volume of 0.5 ml. for a mouse of 20 g.
Statistical analysis. Experimental data were expressed
as a mean ± SD and statistically analyzed using U – t
test of Graph Pad Software. p values 0.05 were con-
sidered significant.
As shown in Table 2, lipoperoxidation significantly
increased (p < 0.01) in the cisplatin-treated animals in
comparison with the non-treated group. No significant
difference (p > 0.05) was found between lipoperoxida-
tion in untreated animals and the boldo infusion-treated
animals. Pretreatment with boldo (P. Boldus) infusion
was found to significantly reduce (p < 0.05) the lipo-
peroxidation induced by a single cis-DDP injection.
The data (Table 3) indicate that lipoperoxidation in
the group pretreated with boldine prior to the cis-
DDP injection significantly (p < 0.05) decreased in
comparison with animals treated only with cis-DDP.
No statistical difference (p > 0.05) was found between
the lipoperoxidation found in untreated animals and
boldine-treated animals. The data (Table 4) indicate
that lipoperoxidation in the group pretreated with
Table 3. Effect of boldine on the lipoperoxidation induced by
cis-DDP in mice liver
Pretreated Treatment (nmol MDA/g tissue)
None None 381.8 ± 75.58
None Cis-DDP 661.8 ± 145.34
None Boldine 448.4 ± 168.99
Boldine Cis-DDP 432.0 ± 168.18
Table 2. Effect of boldo infusion on the lipoperoxidation induced
by cis-DDP in the liver of mice
Pretreated Treatment (nmol MDA/g tissue)
None None 381.8 ± 75.58
None Cis-DDP 661.8 ± 145.34
None Boldo infusion 353.2 ± 125.11
Boldo infusion Cis-DDP 496.8 ± 58.83
Copyright © 2009 John Wiley & Sons, Ltd. Phytother. Res. 23, 1024–1027 (2009)
DOI: 10.1002/ptr
by cis-DDP since under our experimental conditions
mice pretreated with a boldo infusion and then treated
with cis-DDP presented significantly lower lipoper-
oxidation in the hepatic tissue than found in the mouse
group treated only with cis-DDP (p < 0.05).
The action of the bold infusion could be explained
by the presence of boldine and catechin, substances
with recognized antioxidant activity due to their ability
to act as free radical scavengers (Schmeda-Hirschmann
et al., 2003). Our results indicate that lipoperoxidation
is significantly lower in mice pretreated with boldine
and catechin followed by cis-DDP treatment than in
mice only treated with cis-DDP (P < 0.05). Boldine
and catechin activity is attributed to the formation
of phenoxy radical and other free radical species due
to the oxidation of boldine’s aporfinic structure and
catechin’s polyphenolic structure (Ubeda et al., 1993),
which act decomposing superoxide anions, hydrogen
peroxides and hydroxyl radicals. Additionally, boldine
has been shown to restore the activity of some enzymes
that participate in antioxidant systems such as gluta-
thione S – transferase in Hepa – 1 cells (Kubínová
et al., 2001), glutathione peroxidase in rats’ liver mito-
chondria (Jang et al., 2000), enzymes whose activity
diminishes in the liver and kidney after cis-DDP admini-
stration (Sadzuka et al., 1992).
Based on our study’s results and considering that
boldine’s solubility in water is minimal, we propose that
the observed effect for the total boldo leaf infusion
is principally due to the presence of catechin, which is
consistent with the highest free radical scavenging
capacity of the infusion attributed to catechin by other
authors (Schmeda-Hirschmann et al., 2003).
In conclusion, the data obtained in our study allow
us to recommend the use of a complete boldo leaf
infusion as an effective chemoprotector agent due to
its activity that prevents the oxidative damage caused
by cis-DDP at the hepatic level.
Future studies should precisely determine if boldo
infusion’s activity lies only in its capacity to act on free
radicals or if it also includes a restoring activity at the
level of some oxidative enzymes without interfering in
cis-DDP’s antitumoral activity. In this regard we must
inform that we are conducting experiments to know if
the boldo infusion has any preventive effect on genetic
damage induced by cis-DDP. Our preliminary results
indicate that the infusion can prevents genetic damage
induced by this antitumoral drug.
This work was supported by the grant No. 055109 3/R from the
Universidad del Bío-Bío, Chile. The authors are very grateful to
Dr Patricio Peñailillo Brito from the Universidad de Talca for the
identification of plant material and to Mr Gerardo Quezada Silva for
his technical assistance and collection of plant material.
Table 4. Effect of catechin on the lipoperoxidation induced by
cis-DDP in mice liver
Pretreated Treatment (nmol MDA/g tissue)
None None 381.8 ± 75.58
None Cis-DDP 661.8 ± 145.34
None Catechin 495.6 ± 31.67
Catechin Cis-DDP 405.4 ± 129.71
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has been demonstrated to be the principal contributor
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an estimated content of boldine in leaves of around
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and 60.9% of the total antioxidant activity, respectively
(Quezada et al., 2004).
Our study provides the first evidence that a boldo
infusion could prevent the oxidative damage induced
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... Fernández et al. (29) reported that Boldo (Peumus boldus molina) has an important antioxidant effect against oxidative damage caused by cisplatin and reduces the MDA increase caused by cisplatin. Previous studies, it was reported that the MDA level increased with hydrochloric acid administration and serum oxidative parameters changed with cisplatin administration approached normal with dexpanthenol administration. ...
... It has been reported that apoptotic and degenerative disorders may occur with an increase in abnormal sperm count, change in sperm density and motility, especially due to increased oxidative stress. (29). ...
Full-text available
This study was carried out to determine the protective effects of dexpanthenol and boldin against possible testicular damage caused by cisplatin. 48 Male Wistar rats (250-270 g) aged 4-5 months were used in the study. 8 groups were formed with 6 animals in each group. Boldin (20 and 40 mg/kg) and dexpentanol (250 and 500 mg/kg) were administered to rats along with cisplatin (7 mg/kg). At the end of the 14th day, spermatogenic and histopathological examinations and testicular tissue malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) levels were determined. A significant decrease in sperm quality and motility, a significant increase in MDA, and a significant decrease in SOD, GSH-Px, and CAT levels were observed in the cisplatin group. MDA levels were significantly lower in the other groups compared to the cisplatin group. SOD, GSH-Px and CAT enzyme levels increased in the treatment groups compared to the cisplatin group and approached the control group. Combined application of high doses of both substances with cisplatin reduced histopathological lesions. Significant increases were found in the groups in which both doses of dexpanthenol and boldin were combined with cisplatin compared to the cisplatin group. As a result, it was concluded that dexpanthenol and boldin treatments may have a protective effect against cisplatin-induced testicular damage in rats.
... Prevents the oxidative hepatic damage induced by cisplatin. [162] AGS cell Helicobacter pylori Inhibition Helicobacter pylori adhesion to human gastric cells. ...
... [122,161] In an in vivo study, a boldo infusion reduced the oxidative damage induced in mice. [162] Another study suggested that boldo extract has a potent anti-Helicobacter pylori properties, directly linked to the presence of catechin-derived PAs. [163] One study showed the antioxidant and anti-diabetic activities (i.e. ...
Certain countries have the privilege of diverse ecosystems that allow access to wide food availability. This fact carries an intrinsic diversity in bioactive compounds such as phytochemicals, especially polyphenols. The aim of this review is to summarize the advances in polyphenols research which have been conducted in Chile, with a focus on polyphenol-rich foods and health-related outcomes. In the first part, several studies that have analyzed food sources rich in polyphenols are presented. This is followed by a description of in vitro and in vivo studies from Chile that have evaluated the polyphenol compounds of Chilean foods or their extracts along with their biological activity or health effects. Most polyphenol studies in our search have an in vitro experimental design where mainly protective activities are tested. The antioxidant effect is remarkable in all studies. As well as discussing the future direction of dietary assessment and the approach to biomarkers in this field, currently, additional emphasis and research investment are necessary to explore more native foods with an added value. Abbreviations: TPC: total Phenolic Content; PEE: phenolic-enriched extract; PAs: proanthocyanidins; 24 HDR: 24-hour dietary recall; FFQ: food frequency questionnaires.
... It is known for its properties for liver treatment, due to boldine [5]. Boldo leaves have been shown to possess more than 30 compounds such as quercetin glycosides, kaempferol derivatives, phenolic acids, proanthocyanidins, among others [6,7]. In this study [8], they compared the content of phenolic compounds, antioxidant capacity and anti-inflammatory activity of commercial herbal infusions, and determined that boldo infusions had a higher content of phenolic compounds and antioxidant capacity than other samples. ...
The aim of this study was to obtain an extract from boldo leaves (Peumus boldus) with antioxidant properties, studying its bioaccessibility and potential application in the development of orange biscuits. The extraction conditions were defined by performing a factorial design, working with two variable factors: extraction time and solvent concentration (96° ethanol and water). Total phenolic compounds (TPC) content was determined by Folin-Ciocalteau method, and antioxidant capacity (AC) against ABTS•+ radicals. On the selected extract, phenolic compounds were identified by UHPLC. To evaluate its application as a functional ingredient, TPC and AC were determined to orange biscuits with different extract content. Furthermore, bioaccessibility was determined by applying the standardized static in vitro digestion method Infogest. Results showed that 1 h extraction time, using water as solvent at 70 °C had the highest content of total phenolic compounds (218.83 ± 25.91 mg eq GAE/g of extract) and the best antioxidant potential being the AC of 720.56 ± 15.00 μMol eq TE/g of extract. Gallic acid, chlorogenic acid, rutin and catechin were identified as the main phenolic compounds on this sample. Regarding the use of boldo extract in the development of a functional orange biscuits, a significant increase in the content of TPC and AC was observed compared to a control biscuit (without extract addition). The study of bioaccessibility evi-denced a reduction of 87% in the content of phenolic compounds. Thereby, further studies in strategies of encapsulation of the extract are needed. In conclusion, boldo extract was found to be an ingredient with potential functionality for its use in food processing.
... Cancer chemotherapeutic agents are notorious for their systemic toxicity, and one might expect antioxidants to alleviate some of these unwanted effects. In fact, aqueous and ethanolic boldo extracts exert a cytoprotective action, both in vitro and in vivo, against a cisplatin challenge (Fernández, Lagos, Rivera, & Zamorano-Ponce, 2009;Mondal, Bishayee, Panigrahi, & Khuda-Bukhsh, 2014). The administration of either free or nanoencapsulated boldine at optimal concentrations of 190 and 80 μM respectively, though weakly cytotoxic, increased the viability of normal liver cells treated with 20 μg/mL cisplatin without improving the survival of hepatocyte carcinoma cells (Mondal, Panigrahi, & Khuda-Bukhsh, 2015). ...
Boldine is an aporphine (isoquinoline) alkaloid found in many angiosperms, particularly the more “primitive” families. Despite its wide distribution, it is seldom abundant and the only commercial source is the bark of the Chilean boldo tree (Peumus boldus Mol.). Due to the medicinal use of boldo leaf infusions in Chile and later in numerous other countries, an early chemical study led to the discovery of boldine followed by the study of its pharmacological properties. In line with its traditional use as an aid to digestion, it was first shown to increase bile flow, and later on to have strong antioxidant and antiinflammatory properties and to antagonize adrenergic and dopaminergic neurotransmission. More recently a plethora of novel pharmacological actions has been revealed, initiating several highly promising research avenues with high potential for clinical development.
... On a Drosophila melanogaster wing-spot test, leaf extracts were not mutagenic, but were able to prevent the mutagenic effect of ethyl methane sulphonate (Carmona et al., 2017). Finally, P. boldus infusion was able to reduce lipoperoxidation caused by the drug cisplatin in a Mus musculus liver model (Fernández et al., 2009). ...
Peumus boldus is an endemic tree species from Chile whose leaves have been the focus of study for decades given that their infusions are reported to relieve rheumatic symptoms, headache, dyspepsia, urinary tract inflammation, and symptoms of other illnesses. These health properties have been studied mainly using leaves and bark, then it is relevant to know more about these properties in different parts of the plant. Considering the importance of P. boldus fruits in the diet of some rural populations, we analyzed their properties to explore its impact on the Chilean population health. Liquid chromatography and mass spectrometry analysis confirmed the presence of alkaloids such as boldine, although aporphine N-methyl-laurotetanine was the most abundant. In addition, flavonoids catechin, chrysin and quercetin were also found in the extract. Cytotoxicity and anti-inflammatory activities of the fruit extract were in-vitro tested by using a murine macrophage cell model, observing that a diluted fraction of the extract was not cytotoxic, but showed anti-inflammatory activity, which is likely attributed to antioxidants activities. By means of quantum chemical calculations, we calculated the redox potential of the respective alkaloids and flavonoids found in the extract. Results suggest a synergistic effect between alkaloids and flavonoids, where boldine and N-methyl-laurotetanine showed similar antioxidant properties. Finally, we present a description of the oxidation mechanisms for both groups of molecules which will sustain P. boldus fruit biological properties, in order to give this kind of fruits scientific value focusing on human health.
... Peumus boldus Mol., (Monimiaceae) is a Chilean medicinal tree used for gastrointestinal and liver diseases [1,2]. In Chile, this tree also is called Boldo, Peta, Voldu or Boldu and botanically was described for the first time by Molina in 1782. ...
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Peumus boldus Mol., is a Chilean medicinal tree used for gastrointestinal and liver diseases. Such medicinal properties are associated with the presence of bioactive flavonoids and aporphine alkaloids. In this study, a new green and efficient extraction method used seven natural deep eutectic solvents (NADES) as extraction media. The extraction efficiency of these NADES was assessed, determining the contents of boldine and total phenolic compounds (TPC). Chemical profiling of P. boldus was done by high-performance liquid chromatography coupled to photo diode array detector and electrospray ion-trap mass spectrometry (HPLC-PDA-ESI-IT/MS) and electrospray ionization quadrupole time-of-flight high-resolution mass spectrometry (HPLC-ESI-QTOF-MS). Among the NADES tested, NADES4 (choline chloride-lactic acid) and NADES6 (proline-oxalic acid) enable better extraction of boldine with 0.427 ± 0.018 and 2.362 ± 0.055 mg of boldine g-1 of plant, respectively. Extraction of boldine with NADES4 and NADES6 was more efficient than extractions performed with methanol and water. On the other hand, the highest TPC were obtained using NADES6, 179.442 ± 3.79 mg of gallic acid equivalents (GAE g-1). Moreover, TPC in extracts obtained with methanol does not show significant differences with NADES6. The HPLC-PAD-MS/MS analysis enable the tentative identification of 9 alkaloids and 22 phenolic compounds. The results of this study demonstrate that NADES are a promising green extraction media to extract P. boldus bioactive compounds and could be a valuable alternative to classic organic solvents.
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Background Synthetic glucocorticoid therapeutic agent methylprednisolone (MPL), when used for an extended period of time at high dose, promotes the development of reactive oxygen species (ROS)-induced liver toxicity. This study investigated the role of boldine, a natural antioxidant with anti-apoptotic and anti-inflammatory properties, against MPL-induced hepatoxicity in male Wistar rats. Methods 120 rats were divided into eight equal groups: G1 (control), G2, 3, and 4 (rats orally administered 5, 10, and 50 mg boldine/kg b.w./day; respectively, for 28 days), G5 (rats intramuscularly injected with 100 mg MPL/kg b.w. only on the last three days), G6, 7, and 8 (rats administered boldine + MPL). After the last MPL injection, rats were sacrificed at intervals of 1, 24, and 48 h. Results There was a significant decrease in WBCs, RBCs count, and HGB levels, as well as an increase in PLT count, ALT, AST, TG, and LDL levels, and a decrease in HDL level in serum. Oxidative stress markers levels increased at all times, and gene expression of antioxidant enzymes increased at 24h. Immunohistochemical analysis revealed that cytochrome c levels significantly increased after MPL treatment. The COMET assay revealed detectable DNA lesions. There was no immune reactivity of IL-6 expressions as an inflammatory response marker. Conclusions Oral administration of boldine has a modulatory protective, antioxidant, and anti-apoptotic effect against free radicals.
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Este trabalho visou realizar um levantamento etnobotânico das plantas comercializadas como medicinais e fitoterápicos no município de Resende, RJ. Para tanto, foram mapeados os principais tipos de comércio de plantas medicinais e fitoterápicos e as espécies vegetais comercializadas no município foram registradas. O uso tradicional relatado foi correlacionado com as propriedades biológicas dos metabólitos biossintetizados pelas espécies vegetais já descritas na literatura. O levantamento etnobotânico e a pesquisa acerca do uso de espécies vegetais e fitoterápicos no município revelou que 44% das espécies descritas na RENISUS são comercializadas nas feiras e ervanários visitados e, 77% dessas espécies estão entre as mais consumidas. Foi possível observar que muitas das propriedades biológicas para as quais as espécies vegetais são tradicionalmente consumidas têm correlação com os metabólitos biossintetizados e comprovação descrita na literatura. Tais resultados podem auxiliar na implementação da Política Nacional de Plantas Medicinais e Fitoterápicos no munícipio e no Sul Fluminense, além de contribuir para o acesso seguro, o uso racional e sustentável da biodiversidade.
Introduction The use of herbal medicine has been increasing in many parts of the world, despite safety issues. Cerebral palsy (CP) is traditionally considered a group of disorders, which include cognitive, communicative and epilepsy symptoms. Individuals with CP use conventional drugs in addition to herbal medicines, with the expectation of alleviating side effects. However, improper use of herbs may lead to treatment failure or pharmacokinetic interactions. This research investigated herbal medicines and conventional drugs; used by individuals with CP. In addition, individual assessments for each participant of this study was used to gather information in order to improve quality of life. Methods This is a descriptive and qualitative study, in which twelve individuals with CP completed a questionnaire about their use of herbal medicine and conventional drugs. Results Individuals with CP reported ten herbal medicines and twelve conventional drugs; pharmacokinetic interactions were described in detail. Conclusion Fortunately, none of the participants reported clinical evidence related to the interaction, but it is not possible to conclude that the risks, in general, are over-estimated. Therefore, we recommend the implementation of a public education campaign with herb-drug interactions, involving health professionals and patients, in order to avoid future problems.
Boldine is the main alkaloid of Peumus boldus Molina, widely used in the traditional medicine for the treatment of digestive disorders. It is a compound with excellent antioxidant and anti-inflammatory properties already described. Despite the widespread use of P. boldus for digestive disorders treatment, the gastroprotective effect of Boldine remains unknown. Considering the need for new approaches to treat gastric ulcers with fewer side effects than current therapy, this study aimed to investigate the gastroprotective effect of Boldine in mice, as well as the mechanisms underlying this effect. The gastroprotective effect of Boldine was evaluated on gastric ulcer induced by 60% ethanol/0.3 M HCl or indomethacin (100 mg/kg) in mice. Histological analysis and the mucin-like glycoprotein content were evaluated in ethanol-ulcerated tissue, as well as, oxidative stress and inflammatory parameters. The mechanisms involved in the effect of Boldine were evaluated by pretreating mice with NEM (a sulfhydryl group chelator, 10 mg/kg, i.p.), l-NAME (a non-selective nitric oxide synthase inhibitor, 70 mg/kg, i.p.), yohimbine (an alpha-adrenergic receptor antagonist, 2 mg/kg, i.p.) and indomethacin (a cyclooxygenase inhibitor, 10 mg/kg, i.p.). In addition, the in vitro effect of Boldine on H⁺/K⁺-ATPase activity was determined. Boldine was able to protect gastric mucosa against the damage induced by ethanol/HCl and indomethacin, as evidenced by reduced lesion area and histological analysis. Moreover, the alkaloid reduced oxidative stress and inflammatory mediators in ethanol-ulcerated tissue, beyond has increased mucin-like glycoprotein amount. Finally, Boldine effect is dependent on non-protein sulfhydryl groups and prostanoids but does not involve the inhibition of H+/K + -ATPase activity, being a promising natural resource for gastric ulcer treatment.
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Possible chemoprotective effects of the naturally occurring alkaloid boldine, a major alkaloid of boldo (Peumus boldus Mol.) leaves and bark, including in vitro modulations of drug-metabolizing enzymes in mouse hepatoma Hepa-1 cell line and mouse hepatic microsomes, were investigated. Boldine manifested inhibition activity on hepatic microsomal CYP1A-dependent 7-ethoxyresorufin O-deethylase and CYP3A-dependent testosterone 6 beta-hydroxylase activities and stimulated glutathione S-transferase activity in Hepa-1 cells. In addition to the known antioxidant activity, boldine could decrease the metabolic activation of other xenobiotics including chemical mutagens.
Boldo is an endemic plant from Chile widely used in folk medicine, whose biological (mainly antioxidant) activity has been attributed to the presence of the alkaloid boldine. The aim of this study was to determine the contribution of the flavonoid and other crude fractions from boldo leaves to the antioxidant activity of extracts. The main constituent in the flavonoid fraction was also isolated. The antioxidant activity and reducing power of crude boldo extract and fractions were measured and compared with natural and synthetic antioxidants. The antioxidant activity of boldo leaf extracts came mainly from the flavonoid fraction (44.1%) followed by the alkaloid fraction (15.6%), with catechin and boldine being the main contributors of the antioxidant activity of these 2 fractions (60.9% and 35.6% of the total activity, respectively). On the basis of these results, we propose to use catechin and boldine as markers for the antioxidant activity of boldo leaves. Tailor-made antioxidants to be used in foods can be prepared by enriching these 2 fractions.
Increases in lipid peroxide in kidneys of rats treated with cisplatin were examined in relation to decreases in the activities of Cu,Zn-superoxide dismutase (Cu,Zn-SOD), Mn-SOD, glutathione peroxidase (GSHpx), glutathione S-transferase (GST) and catalase. The activities of catalase, GSHpx and GST in the kidney and the liver were significantly decreased after cisplatin administration. The decrease of GST activity in the kidney was 87.3%, this was the largest decrease among these enzymes in the tissues examined. Cu,Zn-SOD activity significantly decreased only in the kidney. In contrast, the activities of these enzymes in the heart and the lung, which showed no increase in lipid peroxide in our previous study, were not significantly decreased. Cisplatin does not directly increase lipid peroxidation in vitro; therefore, the increase of lipid peroxide in the kidneys of these rats treated with cisplatin can be attributed to a decrease in the activities of lipid peroxide-protecting enzymes.
Lipid peroxidation in biological systems has been considered as one of the major mechanisms of cell injury in aerobic organisms subjected to oxidation stress. Plants, among other functions, are considered to act as free radical scavengers and as antioxidants. Iron II (Fe(2+)), sodium nitroprusside (SNP) and nitropropionic acid stimulate the production of free radicals and lipid peroxidation. In this study, four commonly used tropical medicinal plants (Kigelia africana, Calotropis procera, Hibiscus sabdariffa and Alchornea cordifolia) were studied (in vitro) for their effects on the formation of thiobarbituric acid reactive substances (TBARS) induced by different pro-oxidants (10 microM FeSO(4), 5 microM -sodium SNP and 2mM 3-nitropropionic acid) in rat liver homogenate. All the pro-oxidants significantly increased (P<0.05) the formation of TBARS, which indicates increased lipid peroxidation in the rat liver (in vitro). However, all the plant extracts statistically (P<0.05) reduced the production of TBARS in a concentration-dependent manner in all the tested pro-oxidant-induced oxidative stresses. Alchornea cordifolia appeared to offer the highest protection. The results of the present study suggest that the use of these plants in the treatment of various diseases, especially liver disease, is probably due to their ability to act as antioxidants.
The reaction of lipid peroxides in animal tissues with thiobarbituric acid was dependent on pH of the reaction mixture as was the case for linoleic acid hydroperoxide. The optimum pH was found to be 3.5. Taking this fact into consideration, a standard procedure for the assay of lipid peroxide level in animal tissues by their reaction with thiobarbituric acid was developed as follows. Ten percent ( tissue homogenate was mixed with sodium dodecyl sulfate, acetate buffer (pH 3.5), and aqueous solution of thiobarbituric acid. After heating at 95°C for 60 min, the red pigment produced was extracted with n-butanol-pyridine mixture and estimated by the absorbance at 532nm. As an external standard, tetramethoxy-propane was used, and lipid peroxide level was expressed in terms of nmol malondialdehyde. Using this method, the liped peroxide level in the liver of rats suffering from carbon tetrachloride intoxication was investigated. The results were in good agreement with previously reported data obtained by measuring diene content.
cis-Diamminedichloroplatinum(II) (cisplatin) is one of the most effective antitumor agents currently available for cancer chemotherapy. Unfortunately, its clinical use is severely limited by its dose-dependent nephrotoxicity. Recently, it has been reported that this nephrotoxicity is associated with an increase in lipid peroxides in the rat kidney. However, it is not clear if the specific nephrotoxicity is related to the increase in lipid peroxides induced by cisplatin since the increase in lipid peroxides in the kidney has not been compared with that in other tissues. We examined the changes in lipid peroxide levels in rat tissues after cisplatin administration to clarify the relationship between lipid peroxides and cisplatin nephrotoxicity. After cisplatin administration (5 mg/kg i.p.), the relative weight of the rat liver decreased, but that of the kidney increased. The increase in lipid peroxide levels in the kidney and liver of cisplatin-treated rats was significant. The percent change from control was greatest in the kidney on the 5th day after administration among all tissues examined. Cisplatin did not affect enzymatic or non-enzymatic lipid peroxidation in microsomes of rat kidney or liver in vitro. It appears that the increase in lipid peroxides in the kidney was not a direct consequence of cisplatin administration.
The antioxidant action of a series of benzylisoquinoline alkaloids has been investigated. Laudanosoline, protopapaverine, anonaine, apomorphine, glaucine, boldine, bulbocapnine, tetrahydroberberine and stepholidine produced a dose-dependent inhibition of microsomal lipid peroxidation induced by Fe2+/ascorbate, CCl4/NADPH or by Fe3+ ADP/NADPH. Apomorphine exerted the highest inhibitory effects in the three systems of induction used, with a potency higher than propyl gallate. Laudanosoline was particularly effective in the first system, while bulbocapnine and anonaine were more potent when CCl4/NADPH or Fe3(+)-ADP/NADPH were used as inducers. Laudanosoline, protopapaverine, apomorphine, tetrahydroberberine and stepholidine were also potent inhibitors of nitroblue tetrazolium (NBT) reduction. The presence of a free hydroxyl group or preferably of a catechol group is a feature relevant for inhibition of lipid peroxidation and NBT reduction, nevertheless the antioxidant activity of benzylisoquinoline alkaloids cannot be only ascribed to the formation of phenoxy radicals and other free radical species may be formed during aporphine and tetrahydroprotoberberine oxidation. The influence of this series of compounds on the time course of lipid peroxidation suggests that some of them, like apomorphine and boldine act as chain-breaking antioxidants.
Cisplatin is one of the most active cytotoxic agents in the treatment of cancer, but its clinical use is associated with nephrotoxicity. In the present study we report the effects of different amounts of vitamin C (50, 100 or 200 mg kg(-1)body wt.) in rat kidneys treated with cisplatin (5 mg kg(-1)body wt.), using single doses of both compounds. Cisplatin administration induced lipid peroxidation which was accompanied by a decrease in renal glutathione level in animals killed 7 days after treatments. Furthermore, an increase in serum creatinine has been observed. Treatment of animals with vitamin C 10 min prior to the cisplatin inhibited cisplatin-mediated damage. Seven days after vitamin C plus cisplatin treatments, the depleted level of glutathione and changes in the creatinine clearance recovered to significant levels (P<0.05). Similarly, the enhanced serum creatinine levels which are indicative of renal injury showed a significant reduction (P<0.05) with the three doses of vitamin C tested. The protective effect of vitamin C was dose-dependent. The results suggest that vitamin C is an effective chemoprotective agent against nephrotoxicity induced by the antitumoral cisplatin in Wistar adult rats.
Boldine is a natural compound with well-established free radical scavenger and hepatoprotective properties. The further exploration of its actual therapeutic potential as an antioxidant is, however, partially limited by the absence of knowledge on its pharmacokinetics. In the present studies, we provide information on the in vitro and in vivo biological disposition of boldine. The addition of 200 microM boldine to an isolated rat hepatocyte suspension was followed by a time-dependent (0-60 min) disappearance of boldine from the extracellular medium. This decline was associated with an early (first 2 min) and swift accumulation (1600 microM) of boldine within the cells. Although the intracellular concentration of boldine diminished, boldine was always found to occur within the cells at concentrations substantially higher than those initially added to the preparation. Boldine was also concentration-dependently removed from the extracellular medium by isolated rat livers portally perfused with the antioxidant. In vivo studies, conducted in rats, revealed that following either its oral or its intravenous administration, plasma boldine concentrations declined rapidly and according to an apparently first order type of kinetics. After its oral administration (50 or 75 mg/kg), boldine was rapidly (within 30 min) absorbed and preferentially concentrated in the liver, with substantially lower concentrations being found in the brain and heart. Maximal hepatic concentrations of boldine were found to be equal to or greater than those needed to afford antioxidant and hepatoprotective effects in vitro.
Increased oxidative stress has been suggested to be involved in the pathogenesis and progression of diabetic tissue damage. Several antioxidants have been described as beneficial for oxidative stress-associated diseases. Boldine ([s]-2,9-dihydroxy-1, 10-dimethoxyaporphine) is a major alkaloid found in the leaves and bark of boldo (Peumus boldus Molina), and has been shown to possess antioxidant activity and anti-inflammatory effects. From this point of view, the possible anti-diabetic effect of boldine and its mechanism were evaluated. The experiments were performed on male rats divided into four groups: control, boldine (100 mg kg(-1), daily in drinking water), diabetic [single dose of 80 mg kg(-1)of streptozotocin (STZ), i.p.] and diabetic simultaneously fed with boldine for 8 weeks. Diabetic status was evaluated periodically with changes of plasma glucose levels and body weight in rats. The effect of boldine on the STZ-induced diabetic rats was examined with the formation of malondialdehydes and carbonyls and the activities of endogenous antioxidant enzymes (superoxide dismutase and glutathione peroxidase) in mitochondria of the pancreas, kidney and liver. The scavenging action of boldine on oxygen free radicals and the effect on mitochondrial free-radical production were also investigated. The treatment of boldine attenuated the development of hyperglycemia and weight loss induced by STZ injection in rats. The levels of malondialdehyde (MDA) and carbonyls in liver, kidney and pancreas mitochondria were significantly increased in STZ-treated rats and decreased after boldine administration. The activities of mitochondrial manganese superoxide dismutase (MnSOD) in the liver, pancreas and kidney were significantly elevated in STZ-treated rats. Boldine administration decreased STZ-induced elevation of MnSOD activity in kidney and pancreas mitochondria, but not in liver mitochondria. In the STZ-treated group, glutathione peroxidase activities decreased in liver mitochondria, and were elevated in pancreas and kidney mitochondria. The boldine treatment restored the altered enzyme activities in the liver and pancreas, but not the kidney. Boldine attenuated both STZ- and iron plus ascorbate-induced MDA and carbonyl formation and thiol oxidation in the pancreas homogenates. Boldine decomposed superoxide anions, hydrogen peroxides and hydroxyl radicals in a dose-dependent manner. The alkaloid significantly attenuated the production of superoxide anions, hydrogen peroxide and nitric oxide caused by liver mitochondria. The results indicate that boldine may exert an inhibitory effect on STZ-induced oxidative tissue damage and altered antioxidant enzyme activity by the decomposition of reactive oxygen species and inhibition of nitric oxide production and by the reduction of the peroxidation-induced product formation. Boldine may attenuate the development of STZ-induced diabetes in rats and interfere with the role of oxidative stress, one of the pathogeneses of diabetes mellitus.