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1.
Int J Mol Sci ; 25(1)2024 Jan 03.
Article in English | MEDLINE | ID: mdl-38203800

ABSTRACT

Tendinopathy (TP) is a complex clinical syndrome characterized by local inflammation, pain in the affected area, and loss of performance, preceded by tendon injury. The disease develops in three phases: Inflammatory phase, proliferative phase, and remodeling phase. There are currently no proven treatments for early reversal of this type of injury. However, the metabolic pathways of the transition metabolism, which are necessary for the proper functioning of the organism, are known. These metabolic pathways can be modified by a number of external factors, such as nutritional supplements. In this study, the modulatory effect of four dietary supplements, maslinic acid (MA), hydroxytyrosol (HT), glycine, and aspartate (AA), on hepatic intermediary metabolism was observed in Wistar rats with induced tendinopathy at different stages of the disease. Induced tendinopathy in rats produces alterations in the liver intermediary metabolism. Nutraceutical treatments modify the intermediary metabolism in the different phases of tendinopathy, so AA treatment produced a decrease in carbohydrate metabolism. In lipid metabolism, MA and AA caused a decrease in lipogenesis at the tendinopathy and increased fatty acid oxidation. In protein metabolism, MA treatment increased GDH and AST activity; HT decreased ALT activity; and the AA treatment does not cause any alteration. Use of nutritional supplements of diet could help to regulate the intermediary metabolism in the TP.


Subject(s)
Musculoskeletal Diseases , Oleanolic Acid/analogs & derivatives , Phenylethyl Alcohol/analogs & derivatives , Tendinopathy , Rats , Animals , Rats, Wistar , Dietary Supplements , Lipid Metabolism , Tendinopathy/etiology , Aspartic Acid
2.
Int J Mol Sci ; 23(13)2022 Jun 24.
Article in English | MEDLINE | ID: mdl-35806065

ABSTRACT

Hydroxytyrosol (HT), the main representative of polyphenols of olive oil, has been described as one of the most powerful natural antioxidants, also showing anti-inflammatory, antimicrobial, cardioprotective and anticancer activity in different type of cancers, but has been little studied in hematological neoplasms. The objective of this work was to evaluate the anticancer potential of HT in acute human leukemia T cells (Jurkat and HL60) and the anti-inflammatory potential in murine macrophages (Raw264.7). For this, cytotoxicity tests were performed for HT, showing IC50 values, at 24 h, for Jurkat, HL60 and Raw264.7 cells, of 27.3 µg·mL-1, 109.8 µg·mL-1 and 45.7 µg·mL-1, respectively. At the same time, HT caused cell arrest in G0/G1 phase in both Jurkat and HL60 cells by increasing G0/G1 phase and significantly decreasing S phase. Apoptosis and cell cycle assays revealed an antiproliferative effect of HT, decreasing the percentage of dividing cells and increasing apoptosis. Furthermore, HT inhibited the PI3K signaling pathway and, consequently, the MAPK pathway was activated. Inflammation tests revealed that HT acts as an anti-inflammatory agent, reducing NO levels in Raw264.7 cells previously stimulated by lipopolysaccharide (LPS). These processes were confirmed by the changes in the expression of the main markers of inflammation and cancer. In conclusion, HT has an anticancer and anti-inflammatory effect in the cell lines studied, which were Raw264.7, Jurkat, and HL60, and could be used as a natural drug in the treatment of liquid cancers, leukemias, myelomas and lymphomas.


Subject(s)
Chaperonin 60/metabolism , Olea , Animals , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Apoptosis , Humans , Inflammation/drug therapy , Mice , Phenylethyl Alcohol/analogs & derivatives , Phosphatidylinositol 3-Kinases , Polyphenols/pharmacology , Polyphenols/therapeutic use , Proto-Oncogene Proteins c-akt , Signal Transduction
3.
Molecules ; 27(7)2022 Apr 05.
Article in English | MEDLINE | ID: mdl-35408740

ABSTRACT

There is currently a worldwide consensus and recognition of the undoubted health benefits of the so-called Mediterranean diet, with its intake being associated with a lower risk of mortality. The most important characteristics of this type of diet are based on the consumption of significant amounts of fruit, vegetables, legumes, and nuts, which provide, in addition to some active ingredients, fiber and a proportion of vegetable protein, together with extra virgin olive oil (EVOO) as the main sources of vegetable fat. Fish and meat from poultry and other small farm animals are the main sources of protein. One of the main components, as already mentioned, is EVOO, which is rich in monounsaturated fatty acids and to a lesser extent in polyunsaturated fatty acids. The intake of this type of nutrient also provides an important set of phytochemicals whose health potential is widely spread and agreed upon. These phytochemicals include significant amounts of anthocyanins, stilbenes, flavonoids, phenolic acids, and terpenes of varying complexities. Therefore, the inclusion in the diet of this type of molecules, with a proven healthy effect, provides an unquestionable preventive and/or curative activity on an important group of pathologies related to cardiovascular, infectious, and cancerous diseases, as well as those related to the metabolic syndrome. The aim of this review is therefore to shed light on the nutraceutical role of two of the main phytochemicals present in Olea europaea fruit and leaf extracts, polyphenols, and triterpenes, on healthy animal growth. Their immunomodulatory, anti-infective, antioxidant, anti-aging, and anti-carcinogenic capabilities show them to be potential nutraceuticals, providing healthy growth.


Subject(s)
Anti-Infective Agents , Antineoplastic Agents , Olea , Triterpenes , Animals , Anthocyanins/analysis , Anti-Infective Agents/analysis , Anti-Infective Agents/pharmacology , Antineoplastic Agents/analysis , Antioxidants/chemistry , Dietary Supplements , Fruit/chemistry , Olea/chemistry , Olive Oil/chemistry , Phytochemicals/analysis , Plant Extracts/chemistry , Polyphenols/chemistry , Triterpenes/analysis , Triterpenes/pharmacology , Vegetables
4.
Molecules ; 25(17)2020 Sep 03.
Article in English | MEDLINE | ID: mdl-32899159

ABSTRACT

Maslinic acid (MA) is a natural triterpene from Olea europaea L. with multiple biological properties. The aim of the present study was to examine MA's effect on cell viability (by the MTT assay), reactive oxygen species (ROS levels, by flow cytometry) and key antioxidant enzyme activities (by spectrophotometry) in murine skin melanoma (B16F10) cells compared to those on healthy cells (A10). MA induced cytotoxic effects in cancer cells (IC50 42 µM), whereas no effect was found in A10 cells treated with MA (up to 210 µM). In order to produce a stress situation in cells, 0.15 mM H2O2 was added. Under stressful conditions, MA protected both cell lines against oxidative damage, decreasing intracellular ROS, which were higher in B16F10 than in A10 cells. The treatment with H2O2 and without MA produced different responses in antioxidant enzyme activities depending on the cell line. In A10 cells, all the enzymes were up-regulated, but in B16F10 cells, only superoxide dismutase, glutathione S-transferase and glutathione peroxidase increased their activities. MA restored the enzyme activities to levels similar to those in the control group in both cell lines, highlighting that in A10 cells, the highest MA doses induced values lower than control. Overall, these findings demonstrate the great antioxidant capacity of MA.


Subject(s)
Antioxidants/pharmacology , Embryo, Mammalian/cytology , Hydrogen Peroxide/toxicity , Melanoma, Experimental/pathology , Triterpenes/pharmacology , Animals , Cell Death/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Fluorescence , Inhibitory Concentration 50 , Membrane Potential, Mitochondrial/drug effects , Mice , Rats , Triterpenes/chemistry
5.
Phytomedicine ; 23(12): 1301-1311, 2016 Nov 15.
Article in English | MEDLINE | ID: mdl-27765349

ABSTRACT

BACKGROUND: Metabolic syndrome is a set of pathologies among which stand out the obesity, which is related to the lipid droplet accumulation and changes to cellular morphology regulated by several molecules and transcription factors. Maslinic acid (MA) is a natural product with demonstrated pharmacological functions including anti-inflammation, anti-tumor and anti-oxidation, among others. PURPOSE: Here we report the effects of MA on the adipogenesis process in 3T3-L1 cells. METHODS: Cell viability, glucose uptake, cytoplasmic triglyceride droplets, triglycerides quantification, gene transcription factors such as peroxisome proliferator-activated receptor γ (PPARγ) and adipocyte fatty acid-binding protein (aP2) and intracellular Ca2+ levels were determined in pre-adipocytes and adipocytes of 3T3-L1 cells. RESULTS: MA increased glucose uptake. MA also decreased lipid droplets and triglyceride levels, which is in concordance with the down-regulation of PPARγ and aP2. Finally, MA increased the intracellular Ca2+ concentration, which could also be involved in the demonstrated antiadipogenic effect of this triterpene. CONCLUSION: MA has been demonstrated as potential antiadipogenic compound in 3T3-L1 cells.


Subject(s)
Adipocytes/drug effects , Adipogenesis/drug effects , Cell Differentiation/drug effects , Olea/chemistry , Triterpenes/pharmacology , 3T3-L1 Cells , Animals , Calcium/metabolism , Cell Survival/drug effects , Fatty Acid-Binding Proteins/biosynthesis , Fatty Acid-Binding Proteins/genetics , Glucose/metabolism , Mice , PPAR gamma/genetics , RNA/biosynthesis , RNA/genetics , Triglycerides/metabolism , Triterpenes/chemistry
6.
Article in English | MEDLINE | ID: mdl-27178358

ABSTRACT

NADPH plays a central role in reductive biosynthesis of membrane lipids, maintenance of cell integrity, protein synthesis and redox balance maintenance. Hence, NADPH is involved in the growth and proliferation processes. In addition, it has been shown that changes in nutritional conditions produced changes in NADPH levels and growth rate. Maslinic acid (MA), a pentacyclic triterpene of natural origin, is able to stimulate NADPH production, through regulation of the two oxidative phase dehydrogenases of the pentose phosphate pathway. Our main objective was to study the effects of MA on the kinetic behaviour and on the molecular expression of two NADPH-generating systems, NADP-dependent isocitrate dehydrogenase (NADP-IDH) and malic enzyme (ME), in the liver and white muscle of gilthead sea bream (Sparus aurata). Four groups of 12g of a mean body mass were fed for 210days in a fish farm, with diets containing 0 (control), and 0.1g of MA per kg of diet. Two groups were fed ad libitum (C-AL and MA-AL) and another's two, with restricted diet of 1% of fish weight (C-R and MA-R). Results showed that MA significantly increased the main kinetic parameter of the NADPH-forming enzymes (NADP-IDH and ME). In this sense, specific activity, maximum velocity, catalytic efficiency and activity ratio values were higher in MA conditions than control groups. Moreover, these changes were observed in both feeding regimen, AL and R. Meanwhile, the Michaelis constant changed mainly in groups fed with the MA and restricted diet, these changes are related to the best substrate affinity by enzyme. Moreover, in the Western-blot result, we found that MA increased both protein levels studied, this behaviour being consistent with the regulation of the number of enzyme molecules. All results, indicate that MA, independently of the fed regimen, could potentially be a nutritional additive for fish as it improved the metabolic state of fish, as consequence of increased activity and expression of NADP-IDH and ME enzymes.


Subject(s)
Animal Feed , Fish Proteins/metabolism , Isocitrate Dehydrogenase/metabolism , Liver/drug effects , Malate Dehydrogenase/metabolism , Muscle, Skeletal/drug effects , NADP/metabolism , Sea Bream/metabolism , Triterpenes/pharmacology , Animal Nutritional Physiological Phenomena , Animals , Biomarkers/metabolism , Caloric Restriction , Dose-Response Relationship, Drug , Energy Metabolism/drug effects , Kinetics , Liver/enzymology , Muscle, Skeletal/enzymology , Nutritional Status/drug effects , Sea Bream/growth & development , Up-Regulation
7.
PLoS One ; 11(1): e0146178, 2016.
Article in English | MEDLINE | ID: mdl-26751572

ABSTRACT

Maslinic acid (MA) is a natural triterpene present in high concentrations in the waxy skin of olives. We have previously reported that MA induces apoptotic cell death via the mitochondrial apoptotic pathway in HT29 colon cancer cells. Here, we show that MA induces apoptosis in Caco-2 colon cancer cells via the extrinsic apoptotic pathway in a dose-dependent manner. MA triggered a series of effects associated with apoptosis, including the cleavage of caspases -8 and -3, and increased the levels of t-Bid within a few hours of its addition to the culture medium. MA had no effect on the expression of the Bax protein, release of cytochrome-c or on the mitochondrial membrane potential. This suggests that MA triggered the extrinsic apoptotic pathway in this cell type, as opposed to the intrinsic pathway found in the HT29 colon-cancer cell line. Our results suggest that the apoptotic mechanism induced in Caco-2 may be different from that found in HT29 colon-cancer cells, and that in Caco-2 cells MA seems to work independently of p53. Natural antitumoral agents capable of activating both the extrinsic and intrinsic apoptotic pathways could be of great use in treating colon-cancer of whatever origin.


Subject(s)
Adenocarcinoma/drug therapy , Apoptosis , Colonic Neoplasms/drug therapy , Mutation , Receptors, Death Domain/metabolism , Triterpenes/chemistry , Tumor Suppressor Protein p53/genetics , Adenocarcinoma/genetics , Antineoplastic Agents/chemistry , Caco-2 Cells , Caspase 3/metabolism , Caspase 8/metabolism , Cell Proliferation , Colonic Neoplasms/genetics , Cytochromes c/metabolism , Drug Screening Assays, Antitumor , Flow Cytometry , Humans , Inhibitory Concentration 50 , Membrane Potential, Mitochondrial , Treatment Outcome
8.
Article in English | MEDLINE | ID: mdl-26236377

ABSTRACT

Maslinic acid (MA) is a natural compound whose structure corresponds to a pentacyclic triterpene. It is abundant in the cuticular lipid layer of olives. MA has many biological and therapeutic properties related to health, including antitumor, anti-inflammatory, antimicrobial, antiparasitic, antihypertensive, and antioxidant activities. However, no studies have been performed to understand the molecular mechanism induced by this compound in melanoma cancer. The objective of this study was to examine the effect of MA in melanoma (B16F10) cells grown in the presence or absence of fetal bovine serum (FBS). We performed cell proliferation measurements, and the reactive oxygen species (ROS) measurements using dihydrorhodamine 123 (DHR 123) and activities of catalase, glucose 6-phosphate dehydrogenase, glutathione S-transferase, and superoxide dismutase. These changes were corroborated by expression assays. FBS absence reduced cell viability decreasing IC50 values of MA. The DHR 123 data showed an increase in the ROS level in the absence of FBS. Furthermore, MA had an antioxidant effect at lower assayed levels measured as DHR and antioxidant defense. However, at higher dosages MA induced cellular damage by apoptosis as seen in the results obtained.

9.
J Proteomics ; 83: 15-25, 2013 May 27.
Article in English | MEDLINE | ID: mdl-23499989

ABSTRACT

Maslinic acid (MA) is an anti-tumoural agent which shows potent anti-proliferative properties against the HT29 colon-cancer cells. To shed light upon the active mechanism of MA we have investigated its effects upon the cytoskeleton. We used a proteomics procedure based on two-dimensional gel electrophoresis, mass analysis and peptide mass fingerprinting. The incubation of HT29 cells with MA led to G1 cell-cycle arrest. After 24hours' exposure to 3.7µM (IC50/8) and 30µM (IC50) MA fourteen differentially expressed cytoskeletal proteins could be discerned. One group of these proteins, made up of keratin 2, keratin 8, keratin type II cytoskeletal 8, keratin type I cytoskeletal 9, keratin type I cytoskeletal 18, cytokeratins 18 and 19, and ß-actin, exert a structural function, whilst another group, made up of lamin B1, gelsolin 1, septin 2, villin 1, actin-related protein 2 and moesin, is related to the nucleation of actin and cytoskeleton formation. Changes in the expression of moesin, villin 1 and ß-actin identified by the proteomics techniques were corroborated by Western blotting. This is the first evidence obtained of the regulatory effects of MA on the cytoskeleton, which may prove to be one of the bases of its anti-proliferative effect against colon-cancer cells. BIOLOGICAL SIGNIFICANCE: In this paper we describe the changes in the expression of different cytoskeleton proteins identified by the proteomics techniques and corroborated by Western blotting. This is the first evidence obtained of the regulatory effects of MA on the cytoskeleton, which may prove to be one of the bases of its anti-proliferative effect against colon-cancer cells.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Colonic Neoplasms/metabolism , Cytoskeletal Proteins/biosynthesis , Gene Expression Regulation, Neoplastic/drug effects , Neoplasm Proteins/biosynthesis , Triterpenes/pharmacology , Cell Line, Tumor , Cell Proliferation/drug effects , Colonic Neoplasms/drug therapy , Colonic Neoplasms/pathology , G1 Phase Cell Cycle Checkpoints/drug effects , Humans
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