ABSTRACT
Colorectal cancer (CRC) can be either prevented or alleviated using conventional drugs combined with natural treatments. Andean berry (AB, Vaccinium meridionale Sw.) is an underutilized berry with promising anti-inflammatory and antiproliferative effects that could be used to alleviate CRC markers in combination with Aspirin, a well-known CRC preventive drug. This research aimed to evaluate the impact of Aspirin, AB juice (ABJ), and their mixture on colorectal cancer in vitro and in vivo. The treatments (ABJ: 0, 10, 20, and 30 % v/v; Aspirin: 0, 10, 15, and 20 mM; and their combination) were assessed on SW480 cells to test their antiproliferative and pro-apoptotic effect. To evaluate their chemopreventive and chemoprotective effect in vivo, azoxymethane (AOM, 15 mg/kg BW) was used as a chemical inductor of early-stage colon cancer. Balb/c mice (8 weeks' age) were randomly assigned to five groups (n = 6 mice/group): control (no treatment), positive control (AOM-treated mice), AOM + Aspirin (20 mM: 25 mg/kg BW), AOM + ABJ (30 % v/v), and AOM + Aspirin + ABJ (Aspirin: 25 mg/kg BW; ABJ: 30 % v/v). ABJ contained phenolic compounds such as 3,4-dihydroxybenzoic and gallic acids, morin, and rutin. The mixture showed a strongest antiproliferative effect than their counterparts (+10.39-46.23 %). Except for Aspirin (20 mM), the cells were not able to proliferate based on the cloning efficiency test. The mixture was the most effective treatment arresting the cell cycle and increasing G2/M cell population (p < 0.01). Aspirin and ABJ showed mainly intrinsic and extrinsic-mediated apoptotic processes, while the mixture decreased most pro-apoptotic (cytochrome C, DR4, DR5, TNFRSF1A, Bax, and Bad) and anti-apoptotic proteins (Hsp70, Hsp32, and XIAP) compared to the untreated cells. In silico simulations highlighted the interaction between rutin and catalase as the strongest affinity (-10.30 Kcal/mol). ABJ and the mixture decreased aberrant crypt foci in vivo compared to AOM-only treated mice and protected the colonic and liver architecture, this was latter used as a secondary indicator of AOM-metabolic activity. The chemopreventive approach was more effective, related to a prior regulation of cancer-protective mechanisms in vivo, alleviating the AOM-induced damage. The results indicated that Aspirin and ABJ mixtures exhibit antiproliferative and pro-apoptotic effects in SW480 cells inducing mechanisms linked to extrinsic (TNF and TRAIL-mediated apoptosis) and intrinsic (Bax and cytochrome C modulation) pathways. At in vivo levels, the treatments displayed defensive effects against the AOM-induced damage as observed by macroscopic measurements. However, more in vitro, and in vivo approaches are required to completely fulfill the pro-apoptotic, anti-proliferative, and chemopreventive/chemoprotective effects of ABJ.
Subject(s)
Anticarcinogenic Agents , Antineoplastic Agents , Colonic Neoplasms , Vaccinium , Animals , Anticarcinogenic Agents/pharmacology , Antineoplastic Agents/adverse effects , Aspirin/pharmacology , Colonic Neoplasms/chemically induced , Colonic Neoplasms/prevention & control , Cytochromes c , Fruit/metabolism , Mice , Rutin/pharmacology , bcl-2-Associated X ProteinABSTRACT
Andean Berry (Vaccinium meridionale Sw.) is a South American fruit rich in phytochemicals with promising anti-cancer properties as co-adjuvants to nonsteroidal anti-inflammatory drugs such as Aspirin. This study aimed to evaluate the antiproliferative potential of Andean Berry Juice (ABJ) in combination with Aspirin in human SW480 colon adenocarcinoma cells. ABJ primarily contained 3,4-dihydroxybenzoic and chlorogenic acids. The combined treatment of ABJ (IC50 : 30.0 ± 0.11%) and Aspirin (IC50 : 20.0 ± 0.57) exhibited a higher (p < .01) antiproliferative effect than each counterpart. Moreover the same mixture displayed a lower reduced glutathione/oxidized glutathione ratio (GSH/GSSG) than the untreated cells. ABJ-Aspirin combination induced late apoptosis stage without stimulating mitochondrial depolarization and prompted phosphatidylserine relocalization. These results emphasize the antiproliferative potential of bioactive compounds from ABJ and Aspirin combinations. PRACTICAL APPLICATIONS: Natural products such as Andean Berry (V. meridionale Sw.) juice (ABJ) contains antioxidant polyphenols that could reduce the need to use non-steroidal anti-inflammatory drugs, currently employed in cancer treatment, to prevent its side effects. The high abundance of polyphenols from this underutilized berry could stimulate the standardization of its production and industrial exploitation to be transformed into suitable food products delivering natural bioactive compounds with potential anti-cancer effects in vitro.
Subject(s)
Adenocarcinoma , Vaccinium , Adenocarcinoma/drug therapy , Aspirin , Colon , Fruit , HumansABSTRACT
V. meridionale Swartz is an underutilized Andean Berry that has been linked to several health benefits potentially derived from its anti-inflammatory effects. This research aimed to evaluate the impact of Andean Berry Juice (ABJ) combined with Aspirin in the modulation of anti-inflammatory markers from LPS-stimulated RAW 264.7 macrophages. The chemical characterization of ABJ showed a high content of polyphenols, mainly gallic acid (659-75 µg/g) and cyanidin chloride (418.61 µg/mL). Compared to LPS-stimulated macrophages, ABJ, Aspirin, and its combination reduced NO and ROS production (3.26-42.55 and 17.59-65.68%, respectively). In comparison, the half inhibitory concentration of NO reduction (IC50) was found at 7.69% v/v (ABJ) and 24.48 mM (Aspirin). Compared to the pro-inflammatory control (LPS), ABJ reduced IL-1ß, MCP-1, and GCSF; Aspirin decreased IL1R1, MCP-1, GMCSF, GCSF, and TNF-α; and the ABJ + Aspirin treatment reduced IL1R, GMCSF, and CXCL10. The in silico interaction of cytokines and the prediction of potential binding interactions suggested CCR1, CCR5, and NF-kB modulation. These results showed the anti-inflammatory potential of underutilized South American berries and their co-adjuvant effect with known drugs such as Aspirin in the resolution of inflammatory-derived conditions. This is the first report of the anti-inflammatory effects of V. meridionale Swartz juice in combination with Aspirin on LPS-challenged RAW 264.7 macrophages.
Subject(s)
Lipopolysaccharides , Vaccinium , Animals , Anti-Inflammatory Agents/pharmacology , Aspirin/pharmacology , Fruit , Macrophages , MiceABSTRACT
The Neuropeptide EI (NEI, glutamic acid- isoleucine amide) participates in neuroendocrine function. Previously we demonstrated that NEI concentration is regulated by thyroid hormones in discrete hypothalamic areas in rats. We observed that the thyroid status affects the dopaminergic regulation of the pituitary hormones. In this study we explored possible interactions between NEI and tyrosine hydroxylase (TH) containing elements in selected hypothalamic areas of male rats. Neuronal somas, terminals and boutons were assessed by confocal microscopy, in hypo- and hyperthyroid animals. We observed a remodeling of the contacts between the TH and NEI immunoreactive elements in the incerto-hypothalamic area (IHy, also known as rostromedial zona incerta) according to thyroid function. However, in the dorsolateral zone of the peduncular part of the lateral hypothalamus (DL-PLH) the thyroid hormones affect the dendritic trees of the neurons without perturbing the overall NEI/TH contacts. Also, we demonstrated that TRH Receptor 1 (TRH-R1) is colocalized in NEI immunoreactive neurons in the peduncular part of the lateral hypothalamus (PLH) and NEI precursor mRNA expression increased by hypothyroidism indicating that NEI neurons are responsive to the feedback mechanisms of the Hypothalamic Pituitary-Thyroid Axis (HPT). In conclusion, the hypothyroid status seems to increase the interactions between the NEI neurons and the dopaminergic pathways while hyperthyroidism either decreases or displays no effects. Altogether these observations support the participation of the IHy and PLH NEI as a modulating component of the HPT suggesting that altered neuroendocrine, behavioral and cognitive dysfunctions induced by dysthyroidism could be in part mediated by NEI.
Subject(s)
Hyperthyroidism/metabolism , Hypothalamus/metabolism , Hypothyroidism/metabolism , Neuronal Plasticity , Oligopeptides , Tyrosine 3-Monooxygenase , Animals , Hyperthyroidism/enzymology , Hyperthyroidism/physiopathology , Hypothalamus/enzymology , Hypothalamus/physiopathology , Hypothyroidism/enzymology , Hypothyroidism/physiopathology , Male , Neurons/enzymology , Neurons/metabolism , Neurons/physiology , Rats , Rats, WistarABSTRACT
Berry consumption is associated with colorectal-cancer chemoprevention, but digestive conditions can affect this property. The bioaccessibility and apparent permeability coefficients of bioactive compounds from Andean Berry Juice (ABJ) after in vitro gastrointestinal digestion and colonic fermentation were analyzed. The antiproliferative effect of the fermented nondigestible fraction was evaluated against SW480 colon-adenocarcinoma cells. Gallic acid displayed the highest bioaccessibility in the mouth, stomach, small intestine, and colon. However, chlorogenic acid exhibited the highest apparent permeability coefficients (up to 1.98 × 10-4 cm/s). The colonic-fermentation fraction showed an increase of ≥50% antiproliferative activity against SW480 cells (19.32%, v/v), equivalent to those of gallic acid (13.04 µg/g), chlorogenic acid (7.07 µg/g), caffeic acid (0.40 µg/g), ellagic acid (7.32 µg/g), rutin (6.50 µg/g), raffinose (0.14 mg/g), stachyose (0.70 mg/g), and xylose (9.41 mg/g). Bioactive compounds from ABJ are bioaccessible through the gastrointestinal tract and colon fermentation, resulting in antiproliferative activity.
Subject(s)
Colorectal Neoplasms/physiopathology , Fruit and Vegetable Juices/analysis , Plant Preparations/metabolism , Vaccinium/metabolism , Cell Line, Tumor , Cell Proliferation , Colorectal Neoplasms/diet therapy , Colorectal Neoplasms/metabolism , Digestion , Fruit/chemistry , Fruit/metabolism , Gastrointestinal Tract/metabolism , Humans , Plant Preparations/chemistry , Vaccinium/chemistryABSTRACT
Introduction: Vaccinium meridionale Swartz, of the family Ericaceae, is commonly known as mortiño or agraz. The plant is considered a functional food, with a content of anthocyanins and antioxidants similar to or greater than that reported for other Vaccinium species. However, little is known about its nutraceutical and medicinal properties. Objectives: determine the antioxidant activity and cytotoxic and antiproliferative effect of mortiño fruit aqueous extract against colon adenocarcinoma cells (SW480) and their derived metastatic cells (SW620). Methods: total phenols and anthocyanins were determined by the Folin-Ciocalteu method. Caffeoyl derivatives were determined by HPLC-DAD. Antioxidant activity was analyzed as the ability to scavenge reactive oxygen species (ROS), reactive nitrogen species (RNI), peroxyl radicals and hydroxyl radicals. Cytotoxic and antiproliferative activities were studied by MTT and sulforhodamine B. Results: the following substances were found in 100 g of lyophilized extract: total phenols (2 546 mg GAE), anthocyanins (150.7 mg C3G), chlorogenic acid (126 mg), ferulic acid (108 mg) and coumaric acid (63 mg). Hydroxyl radical scavenging capacity was 36 147.5 ?mol DMSO, whereas ROS and RNS scavenging capacity was 29 255.9 y 41 775.2 ?molTrolox, respectively. ORAC value was 41 775.2 ?molTrolox. A dose-dependent cytotoxic and antiproliferative effect was observed. IC50 value was 59.12 µg/ml for SW480 and 56.10 µg/mL for SW620. Conclusions: mortiño fruit aqueous extract exhibited antioxidant, cytotoxic and antiproliferative activities comparable to those of other berries of the genus Vaccinium, which could be partly explained by the presence of a high content of anthocyanins and phenolic acids.
Introducción: Vaccinium meridionale Swartz, pertenece a la familia Ericaceae, conocida comúnmente como mortiño agraz. Considerado un alimento funcional por su contenido de antocianinas y antioxidantes similares o mayor al reportado para otras especies de Vaccinium. Sin embargo, poco se conoce sobre sus propiedades nutracéuticas y en la salud. Objetivos: determinar en extracto acuoso del fruto mortiño, la actividad antioxidante y su efecto citotóxico y antiproliferativo en células de adenocarcinoma de colon (SW480) y sus derivadas metastásicas (SW620). Métodos: contenido de fenoles y antocianinas totales se determinó por el método Folin-Ciocalteu, los derivados caffeoil por HPLC-DAD. La actividad antioxidante se analizó como habilidad para atrapar especies reactivas del oxígeno (ROS) y del nitrógeno (RNI), radicales peróxilo e hidróxilo. La actividad citotóxica y antiproliferativa se estudiaron por MTT y sulforodamina B. Resultados : en 100 g de liofilizado se encontrófenoles totales (2546 mg GAE), antocianinas (150,7 mg C3G), ácido clorogénico (126 mg), ácido ferúlico (108 mg) y cumárico (63 mg), capacidad atrapadora de radical hidroxilo fue 36147,5 ?mol DMSO, capacidad atrapadorade ROS y RNS fue 29255,9 y 41775,2 ?mol Trolox, respectivamente, valor ORAC 41775,2 ?mol Trolox. Se observó un efecto citotóxico y antiproliferativo dosis-dependiente. El valor IC50 para SW480: 59,12 µg/mL y SW620: 56,10 µg/mL. Conclusiones: el extracto acuoso del fruto del mortiño exhibió actividades antioxidantes, citotóxicas y antiproliferativas comparables a las de otras bayas del género Vaccinium, lo que podría ser explicado parcialmente por la presencia del alto contenido de antocianinas y ácidos fenólicos.
ABSTRACT
Flavonoids are polyphenolic compounds able to favour cholesterol-lipid-raft formation and control cell signaling pathways by targeting receptors at the cell surface. Procyanidins (Pcy) are oligomeric and polymeric flavonoids formed by catechins and epicatechins monomers trigger apoptosis by activating TRAIL-death receptors in human colon adenocarcinoma SW480 cells. Here, we investigated whether the apoptotic process triggered by apple procyanidins involving the up-regulation of TRAIL-death receptors DR4/DR5 at the cell surface was dependent on cell membrane lipid-raft formation. We report that Pcy-induced apoptosis was enhanced in presence of nystatin, a cholesterol-sequestering compound inhibiting lipid-raft formation, without changing DR4/DR5 receptor expression. Treatment of SW480 cells with TRAIL caused a 3.5-fold increased level of caveolin together with a 2- to 2.5-fold increased amount of DR4/DR5 proteins in lipid rafts. Pcy-treatment did not induce any alteration in the expression of DR4/DR5 proteins as well as of caveolin present in lipid-raft fractions. Pcy induced an activation of TRAIL-death receptor-mediated apoptosis by a mechanism independent of lipid-raft formation. These results highlight the potential of Pcy as a direct activator of TRAIL-death receptors in cell membrane even in the absence of lipid rafts.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis , Malus/chemistry , Membrane Microdomains/metabolism , Proanthocyanidins/pharmacology , Caveolins/metabolism , Cell Line, Tumor , Humans , Nystatin/pharmacology , Receptors, TNF-Related Apoptosis-Inducing Ligand/metabolism , Receptors, Tumor Necrosis Factor/metabolism , TNF-Related Apoptosis-Inducing Ligand/metabolism , TNF-Related Apoptosis-Inducing Ligand/pharmacologyABSTRACT
On one hand, it has been demonstrated that the exposure of rat brain slices containing caudate putamen and accumbens nuclei to alpha-MSH brings about an increase in cAMP. This increase is affected when dopamine is present in the incubation medium. On the other hand, an interaction of melanotropinergic-like peptides with acetylcholinergic drugs has been showed to be similar to the one observed with dopamine. In this study we have intended to measure cGMP Or IP3 in response to alpha-MSH, and also to study the interaction with cholinergic drugs by measuring the second messengers recently mentioned. cGMP and IP3 have been measured in tissues and medium in their response to the effect of alpha-MSH alone or in the presence of the peptide plus pilocarpine (selective muscarinic agonist) or atropine (selective muscarinic antagonist). None of them modified the cGMP levels when compared with lhe control group. The exposure of rat brain slices containing CP and Acc nuclei to alpha-MSH resulted in an increase in IP3 levels. Pilocarpine by itself brought about an increase of IP3 only when the highest doses was used. Atropine did not modify the IP3 content. However, when slices were exposured to both alpha-MSH and pilocarpine, IP3 content was similar to control values. The blockage of the muscarinic receptor with atropine blocked the IP3 increase induced by alpha-MSH as well. Therefore, we assume that alpha-MSH does not induce changes in cGMP but it does change the IP3 levels, probably acting at the muscarinic receptor level.
Subject(s)
Rats , Animals , Male , alpha-MSH/pharmacology , Atropine/pharmacology , Brain/drug effects , Cyclic GMP/pharmacokinetics , Inositol 1,4,5-Trisphosphate/pharmacokinetics , Pilocarpine/pharmacology , Type C Phospholipases/pharmacology , Rats, WistarABSTRACT
On one hand, it has been demonstrated that the exposure of rat brain slices containing caudate putamen and accumbens nuclei to alpha-MSH brings about an increase in cAMP. This increase is affected when dopamine is present in the incubation medium. On the other hand, an interaction of melanotropinergic-like peptides with acetylcholinergic drugs has been showed to be similar to the one observed with dopamine. In this study we have intended to measure cGMP Or IP3 in response to alpha-MSH, and also to study the interaction with cholinergic drugs by measuring the second messengers recently mentioned. cGMP and IP3 have been measured in tissues and medium in their response to the effect of alpha-MSH alone or in the presence of the peptide plus pilocarpine (selective muscarinic agonist) or atropine (selective muscarinic antagonist). None of them modified the cGMP levels when compared with lhe control group. The exposure of rat brain slices containing CP and Acc nuclei to alpha-MSH resulted in an increase in IP3 levels. Pilocarpine by itself brought about an increase of IP3 only when the highest doses was used. Atropine did not modify the IP3 content. However, when slices were exposured to both alpha-MSH and pilocarpine, IP3 content was similar to control values. The blockage of the muscarinic receptor with atropine blocked the IP3 increase induced by alpha-MSH as well. Therefore, we assume that alpha-MSH does not induce changes in cGMP but it does change the IP3 levels, probably acting at the muscarinic receptor level. (AU)
Subject(s)
Rats , Animals , Male , alpha-MSH/pharmacology , Pilocarpine/pharmacology , Atropine/pharmacology , Inositol 1,4,5-Trisphosphate/pharmacokinetics , Cyclic GMP/pharmacokinetics , Cerebrum/drug effects , Type C Phospholipases/pharmacology , Rats, WistarABSTRACT
Cuando alfa-MSH es inyectado en el área tegmental ventral (VTA) o intracerebroventricularmente (icv) induce comportamiento de aseo excesivo. La infusión icv del péptido tamién provoca el síndrome de estiramiento y bostezo (SEB). Estos efectos son suprimidos por la administración de atropina intraperitoneal, icv o en el ATV. Las evidencias experimentales presentadas sugerían que alfa-MSH actuaría específicamente sobre una aferencia colinérgica en el ATV. Los resultados indicarían que el péptido actuaría en un blanco neural distinto al sistema dopaminérgico , y originaría los cambios comportamentales recientemente mencionados
Subject(s)
Rats , Animals , Male , alpha-MSH/administration & dosage , alpha-MSH/antagonists & inhibitors , alpha-MSH/pharmacology , Atropine/pharmacology , Brain/pathology , Grooming/drug effects , Injections, Intraventricular , Mecamylamine/pharmacology , Motor Activity/drug effectsABSTRACT
Cuando alfa-MSH es inyectado en el área tegmental ventral (VTA) o intracerebroventricularmente (icv) induce comportamiento de aseo excesivo. La infusión icv del péptido tamién provoca el síndrome de estiramiento y bostezo (SEB). Estos efectos son suprimidos por la administración de atropina intraperitoneal, icv o en el ATV. Las evidencias experimentales presentadas sugerían que alfa-MSH actuaría específicamente sobre una aferencia colinérgica en el ATV. Los resultados indicarían que el péptido actuaría en un blanco neural distinto al sistema dopaminérgico , y originaría los cambios comportamentales recientemente mencionados (AU)
