Anticancer Activities of Meroterpenoids Isolated from the Brown Alga Cystoseira usneoides against the Human Colon Cancer Cells HT-29

Colorectal cancer (CRC) is one of the most common types of cancers and a leading cause of cancer death worldwide. The current treatment for CRC mainly involves surgery, radiotherapy, and chemotherapy. However, due to the side effects and the emergence of drug resistance, the search for new anticancer agents, pharmacologically safe and effective, is needed. In the present study, we have investigated the anticancer effects of eight algal meroterpenoids (AMTs, 1-8) isolated from the brown seaweed Cystoseira usneoides and their underlying mechanisms of action using HT-29, a highly metastatic human colon cancer cell line. All the tested meroterpenoids inhibited the growth of HT-29 malignant cells and were less toxic towards non-cancer colon cells, with the AMTs 1 and 5 exhibiting selectivity indexes of 5.26 and 5.23, respectively. Treatment of HT-29 cells with the AMTs 1, 2, 3, 4, 5, and 7 induced cell cycle arrest in G2/M phase and, in some instances, apoptosis (compounds 2, 3, and 5). Compounds 1-8 also exhibited significant inhibitory effects on the migration and/or invasion of colon cancer cells. Mechanistic analysis demonstrated that the AMTs 1, 2, 5, 6, 7, and 8 reduced phosphorylation levels of extracellular signal-regulated kinase (ERK) and the AMTs 2, 3, 4, 5, 7, and 8 decreased phosphorylation of c-JUN N-terminal kinase (JNK). Moreover, the AMTs 1, 2, 3, 4, 7, and 8 inhibited phosphorylation levels of protein kinase B (AKT) in colon carcinoma cells. These results provide new insights into the mechanisms and functions of the meroterpenoids of C. usneoides, which exhibit an anticancer effect on HT-29 colon cancer cells by inducing cell cycle arrest and apoptosis via the downregulation of ERK/JNK/AKT signaling pathways

Meroterpenoids from the Brown Alga Cystoseira usneoides as Potential Anti-Inflammatory and Lung Anticancer Agents

The anti-inflammatory and anticancer properties of eight meroterpenoids isolated from the brown seaweed Cystoseira usneoides have been evaluated. The algal meroterpenoids (AMTs) 1-8 were tested for their inhibitory effects on the production of the pro-inflammatory cytokines tumor necrosis factor (TNF-alpha), interleukin-6 (IL-6), and interleukin-1 beta (IL-1 beta), and the expression of cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) in LPS-stimulated THP-1 human macrophages. The anticancer effects were assessed by cytotoxicity assays against human lung adenocarcinoma A549 cells and normal lung fibroblastic MRC-5 cells, together with flow cytometry analysis of the effects of these AMTs on different phases of the cell cycle. The AMTs 1-8 significantly reduced the production of TNF-alpha, IL-6, and IL-1 beta, and suppressed the COX-2 and iNOS expression, in LPS-stimulated cells (p < 0.05). The AMTs 1-8 displayed higher cytotoxic activities against A549 cancer cells than against MRC-5 normal lung cells. Cell cycle analyses indicated that most of the AMTs caused the arrest of A549 cells at the G2/M and S phases. The AMTs 2 and 5 stand out by combining significant anti-inflammatory and anticancer activities, while 3 and 4 showed interesting selective anticancer effects. These findings suggest that the AMTs produced by C. usneoides may have therapeutic potential in inflammatory diseases and lung cancer

Cytotoxic Activity of Microalgal-derived Oxylipins against Human Cancer Cell lines and their Impact on ATP Levels

Oxylipins are metabolites derived from lipid peroxidation. The plant oxylipin methyl jasmonate (MJ) shows cytotoxic activity against cancer cell lines of various origins, with ATP-depletion being one of the mechanisms responsible for this effect. The cytotoxic activity of oxylipins (OXLs) isolated from the microalgae Chlamydomonas debaryana (13-HOTE) and Nannochloropsis gaditana (15-HEPE) was higher against UACC-62 (melanoma) than towards HT-29 (colon adenocarcinoma) cells. OXLs lowered the ATP levels of HT-29 and UACC-62 cells, but the effect was higher on the second cell line, which had higher basal ATP. This result proves a link between the cytotoxicity and the capability of these compounds to deplete ATP. In addition, the combination of 13-HOTE with the anticancer drug 5-fluorouracil (5-FU) induced a synergistic toxicity against HT-29 cells. These results highlight the therapeutic potential of oxylipins derived from microalgae.España MINECO and FEDER (research project IPT-2011-1370- 060000

Diterpenoids from the Brown Alga Rugulopteryx okamurae and Their Anti-Inflammatory Activity

Brown algae of the Family Dictyotaceae produce an array of structurally diverse terpenoids, whose biomedical potential in the anti-inflammatory area has been scarcely explored. Herein, the chemical study of the alga Rugulopteryx okamurae has led to the isolation of ten new diterpenoids: rugukadiol A (1), rugukamurals A–C (2–4), and ruguloptones A–F (6–10). The structures of the new compounds were established by spectroscopic means. Compound 1 exhibits an unprecedented diterpenoid skeleton featuring a bridged tricyclic undecane system. Compounds 2–10 belong to the secospatane class of diterpenoids and differ by the oxygenated functions that they contain. In antiinflammatory assays, the new diterpenoid 1 and the secospatanes 5 and 10 significantly inhibited the production of the inflammatory mediator NO in LPS-stimulated microglial cells Bv.2 and macrophage cells RAW264.7. Moreover, compounds 1 and 5 were found to strongly inhibit the expression of Nos2 and the pro-inflammatory cytokine Il1b in both immune cell lines

The Algal Meroterpene 11-Hydroxy-11-O-Methylamentadione Ameloriates Dextran Sulfate Sodium-Induced Colitis in Mice

Inflammatory bowel disease (IBD) is a complex class of immune disorders. Unfortunately, a treatment for total remission has not yet been found, while the use of natural product-based therapies has emerged as a promising intervention. The present study was aimed to investigate the anti-inflammatory effects of the algal meroterpene 11-hydroxy-11-O-methylamentadione (AMT-E) in a murine model of dextran sodium sulphate (DSS)-induced colitis. AMT-E was orally administered daily (1, 10, and 20 mg/kg animal) to DSS treated mice (3% w/v) for 7 days. AMT-E prevented body weight loss and colon shortening and effectively attenuated the extent of the colonic damage. Similarly, AMT-E increased mucus production and reduced myeloperoxidase activity (marker for anti-inflammatory activity). Moreover, the algal meroterpene decreased the tumor necrosis factor (TNF)- , interleukin (IL)-1 , and IL-10 levels, and caused a significant reduction of the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Our results demonstrate the protective effects of AMT-E on experimental colitis, provide an insight of the underlying mechanisms of this compound, and suggest that this class of marine natural products might be an interesting candidate for further studies on the prevention/treatment of IBD

Amentadione is a new modulating agent for osteoarthritis in an ex-vivo co-culture preclinical assay

Osteoarthritis (OA) is a whole-joint disease where inflammation interplays with extracellular matrix mineralization in a cycle that leads to its degradation. The lack of effective preventing treatments and disease modifying agents, demands new therapeutic targets and development of effective drugs.FCT: SFRH/BD/111824/2015info:eu-repo/semantics/publishedVersio

Preventive effect of the microalga Chlamydomonas debaryana on the acute phase of experimental colitis in rats

Inflammatory bowel diseases (IBD) are characterised by chronic uncontrolled inflammation of intestinal mucosa. Diet and nutritional factors have emerged as possible interventions for IBD. Microalgae are rich sources of n-3 PUFA and derived oxylipins. Oxylipins are lipid mediators involved in the resolution of many inflammatory disorders. The aim of the present study was to investigate the effects of the oxylipin-containing biomass of the microalga Chlamydomonas debaryana and its major oxylipin constituent, (9Z,11E,13S,15Z)-13-hydroxyoctadeca-9,11,15-trienoic acid ((13S)-HOTE), on acute 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis in rats. Lyophilised microalgal biomass and (13S)-HOTE were administered by oral route 48, 24 and 1 h before the induction of colitis and 24 h later, and the rats were killed after 48 h. The treatment with the lyophilised microalga and (13S)-HOTE improved body-weight loss and colon shortening, as well as attenuated the extent of colonic damage and increased mucus production. Cellular neutrophil infiltration, with the subsequent increase in myeloperoxidase levels induced by TNBS, were also reduced after the administration of the lyophilised microalga or (13S)-HOTE. The anti-inflammatory effects of these treatments were confirmed by the inhibition of colonic TNF-α production. Moreover, lyophilised microalga or (13S)-HOTE down-regulated cyclo-oxygenase-2 and inducible nitric oxide synthase expression. The present study was the first to show the prophylactic effects of a lyophilised biomass sample of the microalga C. debaryana and the oxylipin (13S)-HOTE on TNBS-induced acute colitis in rats. Our findings suggest that the microalga C. debaryana or derived oxylipins could be used as nutraceuticals in the treatment of the active phase of IBD.España Ministerio de Ciencia Innovación, Gobierno de España (grant no. IPT-2011-1370-060000

Amentadione from the Alga Cystoseira usneoides as a Novel Osteoarthritis Protective Agent in an Ex Vivo Co-Culture OA Model

Osteoarthritis (OA) remains a prevalent chronic disease without effective prevention and treatment. Amentadione (YP), a meroditerpenoid purified from the alga Cystoseira usneoides, has demonstrated anti-inflammatory activity. Here, we investigated the YP anti-osteoarthritic potential, by using a novel OA preclinical drug development pipeline designed to evaluate the anti-inflammatory and anti-mineralizing activities of potential OA-protective compounds. The workflow was based on in vitro primary cell cultures followed by human cartilage explants assays and a new OA co-culture model, combining cartilage explants with synoviocytes under interleukin-1 beta (IL-1 beta) or hydroxyapatite (HAP) stimulation. A combination of gene expression analysis and measurement of inflammatory mediators showed that the proposed model mimicked early disease stages, while YP counteracted inflammatory responses by downregulation of COX-2 and IL-6, improved cartilage homeostasis by downregulation of MMP3 and the chondrocytes hypertrophic differentiation factors Col10 and Runx2. Importantly, YP downregulated NF-kappa B gene expression and decreased phosphorylated IkB alpha/total IkB alpha ratio in chondrocytes. These results indicate the co-culture as a relevant pre-clinical OA model, and strongly suggest YP as a cartilage protective factor by inhibiting inflammatory, mineralizing, catabolic and differentiation processes during OA development, through inhibition of NF-kappa B signaling pathways, with high therapeutic potential

Effects of feeding European seabass (Dicentrarchus labrax) juveniles with crude, hydrolysed and fermented biomass of the invasive macroalga Rugulopteryx okamurae (Ochrophyta)

The increasing expansion of the invasive Asian alga Rugulopteryx okamurae along the Andalusian coasts is a major challenge to marine biodiversity, and urgent coordinated measures are required for its removal or elimination. Among the different actions, the biotechnological valorisation could be a strategy for the management of alien biomass within the frame of the Blue Economy. This work evaluates the potential of R. okamurae biomass, either raw or processed by biotechnological treatments based on enzymatic hydrolysis and fermentation processes, to be used as a dietary ingredient in juvenile seabass (Dicentrarchus labrax). These treatments, intended primarily to improve the nutrient vailability, might also decrease the content of undesirable and potentially harmful algal metabolites, such as the diterpenoid dilkamural. To this end, a 90-day feeding trial was carried out using four experimental diets containing 5% (w/w) crude or processed R. okamurae biomass. The results obtained revealed that the inclusion in the diet of raw R. okamurae caused adverse effects on fish growth, yielding lower values than those of the control group (p < 0.05), although this detrimental effect was negligible when the algae was processed prior to its inclusion in the experimental feeds. In terms of muscle composition, diets supplemented with the algal biomass induced an increase in protein content, especially in fish fed with pre-treated biomass. Qualitative differences (p < 0.05) were also found in muscle fatty acid profile, with a significant increase in ARA and DHA contents in fish fed on algae-supplemented diets. A significant reduction (p < 0.05)in muscle and liver lipid oxidation was evidenced in fish fed the algae-enriched diets, especially with treated biomass. Finally, 5% R. okamurae raw biomass caused negative effects on gut functionality, although these effects were not observed when the algal biomass was previously processed. In conclusion, the results revealed the potential of R. okamurae as a functional ingredient for the feeding of juvenile sea bass, provided that the biomass is pre-treated, and corroborated the efficacy of a low-cost biotechnological treatment as a valorisation method for the biomass of this invasive alga

Effects of feeding European seabass (Dicentrarchus labrax) juveniles with crude, hydrolysed and fermented biomass of the invasive macroalga Rugulopteryx okamurae (Ochrophyta)

© 2023 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).The increasing expansion of the invasive Asian alga Rugulopteryx okamurae along the Andalusian coasts is a major challenge to marine biodiversity, and urgent coordinated measures are required for its removal or elimination. Among the different actions, the biotechnological valorisation could be a strategy for the management of alien biomass within the frame of the Blue Economy. This work evaluates the potential of R. okamurae biomass, either raw or processed by biotechnological treatments based on enzymatic hydrolysis and fermentation processes, to be used as a dietary ingredient in juvenile seabass (Dicentrarchus labrax). These treatments, intended primarily to improve the nutrient availability, might also decrease the content of undesirable and potentially harmful algal metabolites, such as the diterpenoid dilkamural. To this end, a 90-day feeding trial was carried out using four experimental diets containing 5% (w/w) crude or processed R. okamurae biomass. The results obtained revealed that the inclusion in the diet of raw R. okamurae caused adverse effects on fish growth, yielding lower values than those of the control group (p < 0.05), although this detrimental effect was negligible when the algae was processed prior to its inclusion in the experimental feeds. In terms of muscle composition, diets supplemented with the algal biomass induced an increase in protein content, especially in fish fed with pre-treated biomass. Qualitative differences (p < 0.05) were also found in muscle fatty acid profile, with a significant increase in ARA and DHA contents in fish fed on algae-supplemented diets. A significant reduction (p < 0.05)in muscle and liver lipid oxidation was evidenced in fish fed the algae-enriched diets, especially with treated biomass. Finally, 5% R. okamurae raw biomass caused negative effects on gut functionality, although these effects were not observed when the algal biomass was previously processed. In conclusion, the results revealed the potential of R. okamurae as a functional ingredient for the feeding of juvenile sea bass, provided that the biomass is pre-treated, and corroborated the efficacy of a low-cost biotechnological treatment as a valorisation method for the biomass of this invasive algae.This research was supported by the Projects VALINVA (Proyecto de Innovación Empresarial con Proyección Territorial en el Ámbito de la Economía Azul CEI⋅MAR (CEIMAR 2019), the Project RUGULOFEED (PCM_00081) (Programa de Ayudas a Proyectos de I+D+i, en régimen de concurrencia competitiva, en el marco del Plan Complementario de Ciencias Marinas y del Plan de Recuperación, Transformación y Resiliencia, Junta de Andalucía) and the Spin-off LifeBioencapsulation S.L. Authors also thank the contribution of the projects BLUEMARO (PID2020-116136RB-I00, Ministerio de Ciencia e Innovación, Spain), PPUENTE2022/007 (Plan Propio de Investigación y Transferencia de la Universidad de Almería), AQUATECH4FEED (grant # PCI2020-112204) granted by MCIN/AEI/10.13039/501100011033 and the EU “NextGenerationEU”/PRTR within the ERA-NET BioBlue COFUND, NEOGIANT (grant # 101036768) granted by the European Union’s Horizon 2020 research and innovation programme. A. Galafat received a Margarita Salas postdoc grant (RR_A_2022_03, Ministerio de Universidades, Spain).All the experimental diets were elaborated by the Universidad de Almería (Experimental feeds Service; http://www.ual.es/stecnicos_spe). Experimental Feeds Service was granted by EQC2018–004984-P and EQC2019–006380-P.Peer reviewe