Antitumor and immunomodulatory activity of fucoidan from the brown alga Lessonia trabeculata (Lessoniaceae) on breast cancer spheroids / Actividad antitumoral e inmunomoduladora de fucoidan del alga parda Lessonia trabeculata (Lessoniaceae) en esferoides de cáncer de mama

Abstract Introduction: The therapeutic benefits of the brown algae fucoidan in the treatment of breast cancer have attracted considerable interest in recent years. However, research using spheroids which provide relevant results in trials for antitumor and immunomodulatory products because they adequately simulate the tumor microenvironment, is limited. Objective: To evaluate the antitumor and immunomodulatory activity of Lessonia trabeculata fucoidan (LtF), native to the Peruvian Sea, on two types of multicellular tumor spheroids. Methods: The study was conducted from January to December 2021. Two types of spheroides were elaborated: from 4T1 tumor cells (MTS), and from 4T1 tumor cells+mouse splenocytes (MTSs). The antitumor activity of LtF was evaluated in MTS by quantifying cell viability with MTT. Immunomodulatory activity was determined in MTSs using the IC50 for two types of treatment: simple, fucoidan alone (LtF) and combined, fucoidan+doxorubicin (LtF+Dox). Pro-inflammatory (TNF-α, IL-6) and anti-inflammatory (IL-10, TGF-β) cytokine production was quantified by sandwich ELISA 72 h after treatment. Dox was used as positive control in all assays. Results: LtF exerted antitumor activity as evidenced by increased necrotic zone and cell debris formation compared to the untreated control. Antitumor activity was concentration dependent between 100 and 6 000 μg/ml. In MTSs, simple treatment increased IL-6 and decreased IL-10 and TGF-β production. The combined treatment significantly reduced TGF-β production. In both treatments and Dox, there was an increase in IL-6 compared to the untreated control. The highest production of IL-10 and TGF-β was observed in the untreated control, compatible with a highly immunosuppressive tumor microenvironment. Conclusions: LtF is a good candidate for the treatment of breast cancer and can immunomodulate the tumor microenvironment alone or in combination with Dox.

Resumen Introduccción: Los beneficios terapéuticos del fucoidan de algas pardas en el tratamiento del cáncer de mama han despertado gran interés en los últimos años. Sin embargo, las investigaciones con esferoides son limitadas, éstos proporcionan resultados relevantes en ensayos de productos antitumorales e inmunomoduladores porque simulan adecuadamente el microambiente tumoral. Objetivo: Evaluar la actividad antitumoral e inmunomoduladora del fucoidan de Lessonia trabeculata (LtF), nativa del Mar Peruano, en dos tipos de esferoides tumorales multicelulares. Métodos: El estudio se realizó de enero a diciembre de 2021. Se elaboraron dos tipos de esferoides: con células tumorales 4T1 (MTS) y con células tumorales 4T1+esplenocitos de ratón (MTSs). La actividad antitumoral de LtF se evaluó en MTS cuantificando la viabilidad celular con MTT. La inmunomodulación se determinó en MTSs utilizando la IC50 para dos tipos de tratamiento: simple, fucoidan solo (LtF) y combinado, fucoidan+doxorubicina (LtF+Dox). La producción de citoquinas proinflamatorias (TNF-α, IL-6) y antiinflamatorias (IL-10, TGF-β) se cuantificó mediante ELISA sándwich 72 h post-tratamiento. En todos los ensayos se utilizó Dox como control positivo. Resultados: En los MTS, el LtF ejerció actividad antitumoral evidenciada por aumento de la zona necrótica y formación de restos celulares respecto al control no tratado. La actividad antitumoral fue concentración-dependiente entre 100 y 6 000 μg/ml. En los MTSs, con el tratamiento simple se incrementó IL-6 y disminuyeron IL-10 y TGF-β. El tratamiento combinado redujo significativamente la producción de TGF-β. Los dos tratamientos y Dox incrementaron IL-6 respecto al control no tratado. La mayor producción de IL-10 y TGF-β se observó en los no tratados, compatible con un microambiente tumoral altamente inmunosupresor. Conclusiones: El LtF es un buen candidato para tratar el cáncer de mama y puede inmunomodular el microambiente tumoral solo o en combinación con Dox.

Research progress in antitumor and immunoregulation of fucoidan / 国际生物医学工程杂志

Fucoidan is a kind of sulfated polysaccharide with various biological activities that mainly exists in the cell walls of brown algae. It is also found in marine invertebrates such as sea cucumbers and sea urchins. Fucoidan has received a lot of attention due to its tumor-killing and immune-boosting properties. Moreover, the combination of fucoidan with chemotherapeutic drugs not only improves antitumor efficacy but also reduces the side effects of these drugs. The function of fucoidan is closely correlated with its structure, molecular weight, degree of sulfation, monosaccharide component, algae source, and time of collection. In this review, the antitumor and immunomodulatory effects of fucoidan are reviewed from the aspects of promoting cell apoptosis, inducing cell cycle arrest, inhibiting angiogenesis and cell migration, and activating immune cells, to provide theoretical guidance for the development and clinical application of fucoidan.

Fucoidan-functionalized activated platelet-hitchhiking micelles simultaneously track tumor cells and remodel the immunosuppressive microenvironment for efficient metastatic cancer treatment

Tumor metastasis is responsible for most mortality in cancer patients, and remains a challenge in clinical cancer treatment. Platelets can be recruited and activated by tumor cells, then adhere to circulating tumor cells (CTCs) and assist tumor cells extravasate in distant organs. Therefore, nanoparticles specially hitchhiking on activated platelets are considered to have excellent targeting ability for primary tumor, CTCs and metastasis in distant organs. However, the activated tumor-homing platelets will release transforming growth factor-β (TGF-β), which promotes tumor metastasis and forms immunosuppressive microenvironment. Therefore, a multitalent strategy is needed to balance the accurate tumor tracking and alleviate the immunosuppressive signals. In this study, a fucoidan-functionalized micelle (FD/DOX) was constructed, which could efficiently adhere to activated platelets through P-selectin. Compared with the micelle without P-selectin targeting effect, FD/DOX had increased distribution in both tumor tissue and metastasis niche, and exhibited excellent anti-tumor and anti-metastasis efficacy on 4T1 spontaneous metastasis model. In addition, due to the contribution of fucoidan, FD/DOX treatment was confirmed to inhibit the expression of TGF-β, thereby stimulating anti-tumor immune response and reversing the immunosuppressive microenvironment. The fucoidan-functionalized activated platelets-hitchhiking micelle was promising for the metastatic cancer treatment.

In vitro cytotoxicity assay of Fucoidan extracted from Turbinaria conoides against cancer cell lines MCF7, A549, and normal cell line L929

Abstract The main aim of the study is to quantify the cytotoxic property of the Fucoidan extracted from the Turbinaria conoides using the MTT assay with the standard fucose. Fucoidan was extracted using the soaked water method and it was determined using the HPLC procedure the obtained Test sample Fucoidan extracted from the Turbinaria conoides and standard fucose was subjected to the cytotoxicity assay against the MCF7 Human breast cancer cell line, A549 lung cancer cell line, and L929 normal mouse fibroblast cell line. From the results it was found that the Test sample showed good IC50 value for MCF7 cell line then A549 with an increasing concentration 24 hours incubation at 37°C The IC50 for MCF7 was 115.21 µg/ml and A549 396.46µg/ml and the Fucoidan extract was checked for its cytotoxicity against the normal mouse fibroblast cell line L929, Fucoidan was found non-lethal to the L929 mouse fibroblast normal cell line. Standard fucose also gave a significant result towards MCF7 and against the L929. This indicates that the Fucoidan extracted from Tubinaria conoides shows better anticancer potential in it. Hence its application can be further extended in the pharmacological fields.

Exploring molecular mechanisms of fucoidan in improving human proximal renal tubular epithelial cells aging by targeting autophagy signaling pathways / 中国中药杂志

Fucoidan(FPS) is an effective component of the Chinese patent medicine named Haikun Shenxi, which treats schronic renal failure in clinics, and has the potential anti-aging effects. However, it is still unclear whether FPS can improve renal aging, especially the molecular mechanism of its anti-aging. The human proximal renal tubular epithelial cells(HK-2) in vitro were divided into normal group(N), D-gal model group(D), low dose of FPS group(L-FPS), high dose of FPS group(H-FPS) and vitamin E group(VE), and treated by the different measures, respectively. More specifically, the HK-2 cells in each group were separately treated by 1 mL of 1% fetal bovine serum(FBS) or D-galactose(D-gal, 75 mmol·L~(-1)) or D-gal(75 mmol·L~(-1))+FPS(25 μg·mL~(-1)) or D-gal(75 mmol·L~(-1))+FPS(50 μg·mL~(-1)) or D-gal(75 mmol·L~(-1))+VE(50 μg·mL~(-1)). After the treatment for 24 h, firstly, the effects of D-gal on senescence-associated β-galactosidase(SA-β-gal) staining characteristics and klotho, P53 and P21 protein expression le-vels, as well as adenosine monophosphate activated protein kinase(AMPK)-uncoordinated 51-like kinase 1(ULK1) signaling pathway activation in the HK-2 cells were detected, respectively. Secondly, the effects of FPS and VE on SA-β-gal staining characteristics and klotho, P53 and P21 protein expression levels in the HK-2 cells exposed to D-gal were investigated, respectively. Finally, the effects of FPS and VE on microtubule-associated protein 1 light chain 3(LC3) protein expression level and AMPK-ULK1 signaling pathway activation in the HK-2 cells exposed to D-gal were examined severally. The results indicated that, for the HK-2 cells, the dose of 75 mmol·L~(-1) D-gal could induce the changes of SA-β-gal staining characteristics and klotho, P53 and P21 protein expression levels. That is causing cells aging. FPS and VE could both ameliorate the changes of SA-β-gal staining characteristics and klotho, P53 and P21 protein expression levels in the HK-2 cells exposed to D-gal. That is anti-cells aging, here, the functions of FPS and VE are similar. D-gal could not only induce cell aging but also increase LC3Ⅱ, phosphorylated-AMPK(p-AMPK) and phosphorylated-ULK1(p-ULK1) protein expressions, and activate autophagy-related AMPK-ULK1 signaling pathway. FPS and VE could both improve the changes of LC3Ⅱ, p-AMPK and p-ULK1 protein expression levels in the HK-2 cells exposed to D-gal. That is inhibiting autophagy-related AMPK-ULK1 signaling pathway activation. On the whole, for the human proximal renal tubular epithelial cells aging models induced by D-gal, FPS similar to VE, can ameliorate renal cells aging by possibly inhibiting autophagy-related AMPK-ULK1 signaling pathway activation. This finding provides the preliminary pharmacologic evidences for FPS protecting against renal aging.

Incremento de la viabilidad, producción de especies reactivas de oxígeno, IL-1 y TNF-α en células mononucleares de sangre periférica humana tratadas con fucoidan de Lessonia trabeculata

Las algas pardas constituyen una fuente de alto contenido de polisacáridos como los fucoidanos que poseen importantes propiedades inmunomoduladoras. El objetivo fue determinar la viabilidad de células mononucleares de sangre periférica humana (CMSPh), producción de óxido nítrico (NO), especies reactivas de oxígeno (ROS) y de las citoquinas proinflamatorias IL-1α, IL-6, TNF-α e IFN-γ en cultivos tratados con fucoidan de Lessonia trabeculata. Se empleó fucoidan de Lessonia trabeculata proveniente de la bahía San Nicolás de Marcona-Ica. Las CMSPh se aislaron empleando Ficoll-Hypaque, se distribuyeron a una concentración de 1x105 células/pocillo en medio RPMI-1640 completo y se trataron con diferentes concentraciones de fucoidan durante 24 y 48 h. La actividad citotóxica se determinó por la reducción de MTT, la producción de NO por la reacción de Griess y las ROS por la reducción del NBT. La producción de citoquinas se cuantificó por ELISA. El fucoidan de L. trabeculata estimuló la proliferación de CMSPh y produjo el incremento de ROS a concentraciones de 100-2000 μg/mL respecto al control (p<0.001), la reacción para nitritos resultó negativa. El fucoidan incrementó la producción de IL-1α y TNF-α a concentraciones de 100 y 10 μg/mL respectivamente, mientras que la producción de IL-6 e IFN-γ no mostró diferencias significativas. Se concluye que el fucoidan de L. trabeculata estimula la proliferación de CMSPh, producción de especies reactivas de oxígeno y las citoquinas proinflamatorias IL-1α y TNF-α que poseen importantes propiedades inmunomoduladoras.

Brown algae are a source of high content of polysaccharides such as fucoidans that have important immunomodulatory properties. The aim was to determine the viability of human peripheral blood mononuclear cells (hPBMC), production of nitric oxide (NO), reactive oxygen species (ROS) and the proinflammatory cytokines IL-1α, IL-6, TNF-α and IFN -γ in cultures treated with fucoidan from Lessonia trabeculata. Fucoidan from Lessonia trabeculata from San Nicolás de Marcona-Ica Bay was used. The hPBMC were isolated using Ficoll-Hypaque, distributed at a concentration of 1x105 cells/well in complete RPMI-1640 medium and treated with different concentrations of fucoidan for 24 and 48 h. The cytotoxic activity was determined by the reduction of MTT, NO production by the Griess reaction and ROS by the reduction of NBT. The production of cytokines was quantified by ELISA. The fucoidan of L. trabeculata stimulated the proliferation of hPBMC and produced the increase of ROS at concentrations of 100-2000 μg/mL with respect to the control (p <0.001), the reaction for nitrites was negative. Fucoidan increased the production of IL-1α and TNF-α at concentrations of 100 and 10 μg/mL respectively, while the production of IL-6 and IFN-γ did not show significant differences. It is concluded that the fucoidan of L. trabeculata stimulates the proliferation of hPBMC, production of reactive oxygen species and the proinflammatory cytokines IL-1α and TNF-α that possess important immunomodulatory properties.

The Safety and Immune-efficacy of Fucoidan from Gagome Kombu (Kjellmaniella crassifolia) on Women with the History of Gynecological Cancer / 日本補完代替医療学会誌

Objective: Fucoidan is a sulfated polysaccharide in brown seaweeds. Previous studies show that fucoidan from Gagome kombu (GKF) exhibit immune-enhancing capabilities on healthy adults. In this study, we focused on the women with the history of gynecological cancer and evaluated the safety and immune-efficacy of GKF.Methods: Ten Japanese women subjects were chosen to ingest the test samples (1 drink / day containing 200 mg GKF) for 4 weeks. Before and after ingestion, blood chemistry analysis, hematological analysis, urinalysis and immune analysis were conducted. Result: Results showed no adverse clinical changes in blood and urinary analysis. In addition, no serious symptoms were observed. Moreover, decrease of serum levels of Eotaxin, IL-7, IL-17 and VEGF were observed.Conclusion: These results indicate that in the case of women with the history of gynecological cancer, GKF is a safe functional food ingredient.

Antioxidant activity of water-soluble polysaccharide extracted from Eucalyptus cultivated in Lebanon

Objective To extract and identify the chemical composition of the polysaccharide isolated from the Eucalyptus cultivated in Lebanon and to evaluate its antioxidant activity. Materials The water-soluble polysaccharide was isolated from Eucalyptus leaves, and its structure was identified by Fourier transform infrared spectroscopy, proton nuclear magnetic resonance, and carbon-13 nuclear magnetic resonance. The antioxidant activity of the active ingredient was screened for its radical scavenging ability using 2,2-diphenyl-1-picryl-hydrazyl radical (DPPH) test. Results The results of the DPPH test have shown that fucoidan, the polysaccharide isolated from Eucalyptus, exhibited almost the same antioxidant activity against DPPH· as the ascorbic acid did at 100 μg/mL. Conclusions This natural molecule extracted from a medicinal plant has a promising antioxidant activity and could be used in pharmaceutical and medical applications.

Protective effect of fucoidin on adriamycin - induced cardiomyocyte cytotoxicity / 中国生化药物杂志

Objective To investigate the protective effect of FUC on adriamycin (ADR) - induced myocardial cell toxicity. Methods Using DPPH FUC method to study the scavenging effect on free radicals play a protective role of H9C2, MTT method is used to study the cytotoxicity of FUC on ADR induced by the H9C2 cell morphological changes were observed by AO/EB staining, the expression of H9C2 in cells of ROS levels and H9C2 cell apoptosis related protein Carboxy-DCFDA Western and Blotting detection method. Results DPPH assay showed that FUC has strong scavenging of oxygen free radicals, FUC to play a good protective effect H9C2 cytotoxicity caused by ADR and mechanism of apoptosis in H9C2 cells were cytotoxic FUC protective effect of H9C2 on ADR induced by the reduced ADR induced increase, elevated levels of reactive oxygen species that caused by ADR the H9C2 cells and H9C2 cells apoptosis protein Caspase-8 Cleaved Cas-pase-3, Cleaved and downreguLation of PARP upreguLation inhibited. Conclusion The protective effect of FUC on myocardial cell toxicity caused by ADR is played by oxidative stress in myocardial cells by antioxidant activity has its own ADR induced inhibition of blocked ADR induced myocardial cell activation of Caspase-8 pathway, thereby effectively inhibit ADR induced Cas-pase-3 activation and PARP shear this continuous process to achieve.

Protective Role of Fucoidan in Cerebral Ischemia-Reperfusion Injury through Inhibition of MAPK Signaling Pathway

Fucoidan has been reported to exhibit various beneficial activities ranging from to antivirus and anticancer properties. However, little information is available about the effects of fucoidan on cerebral ischemia-reperfusion injury (IRI). Our study aimed to explore the effects of fucoidan on cerebral IRI, as well as the underlying mechanisms. Sprague-Dawley (SD) rats were randomly subjected to four groups: Sham, IRI+saline (IRI+S), IRI+80 mg/kg fucoidan (IRI+F80), and IRI+160 mg/kg fucoidan (IRI+F160). Fucoidan (80 mg/kg or 160 mg/kg) was intraperitoneally injected from 7 days before the rats were induced to cerebral IRI model with middle cerebral artery occlusion (MCAO) method. At 24 h after reperfusion, neurological deficits and the total infarct volume were determined. The levels of inflammation-associated cytokines (interleukin (IL)-1β, IL-6, myeloperoxidase (MPO), and tumor necrosis factor (TNF)-α), oxidative stress-related proteins (malondialdehyde (MDA) and superoxide dismutase (SOD)) in the ischemic brain were measured by enzyme-linked immunosorbent assay (ELISA). Besides, the levels of apoptosis-related proteins (p-53, Bax, and B-cell lymphoma (Bcl)-2) and mitogen-activated protein kinase (MAPK) pathway (phosphorylation-extracellular signal-regulated kinase (p-ERK), p-c-Jun N-terminal kinase (JNK), and p-p38) were measured. Results showed that administration of fucoidan significantly reduced the neurological deficits and infarct volume compared to the IRI+S group in a dose-dependent manner. Also, fucoidan statistically decreased the levels of inflammation-associated cytokines, and oxidative stress-related proteins, inhibited apoptosis, and suppressed the MAPK pathway. So, Fucoidan plays a protective role in cerebral IRI might be by inhibition of MAPK pathway.

Fucoidan attenuates 6-hydroxydopamine-induced neurotoxicity by exerting anti-oxidative and anti-apoptotic actions in SH-SY5Y cells

Parkinson's disease (PD) is an irreversible neurological disorder with related locomotor dysfunction and is haracterized by the selective loss of nigral neurons. PD can be experimentally induced by 6-hydroxydopamine (6-OHDA). It has been reported that reactive oxygen species, which deplete endogenous glutathione (GSH) levels, may play important roles in the dopaminergic cell death characteristic of PD. Fucoidan, a sulfated algal polysaccharide, exhibits anti-inflammatory and anti-oxidant actions. In this study, we investigated whether fucoidan can protect against 6-OHDA-mediated cytotoxicity in SH-SY5Y cells. Cytotoxicity was evaluated by using MTT and LDH assays. Fucoidan alleviated cell damage evoked by 6-OHDA dose-dependently. Fucoidan reduced the number of apoptotic nuclei and the extent of annexin-V-associated apoptosis, as revealed by DAPI staining and flow cytometry. Elevation of lipid peroxidation and caspase-3/7 activities induced by 6-OHDA was attenuated by fucoidan, which also protected against cytotoxicity evoked by buthionine-sulfoximine-mediated GSH depletion. Reduction in the glutathione/glutathione disulfide ratio induced by 6-OHDA was reversed by fucoidan, which also inhibited 6-OHDA-induced disruption of mitochondrial membrane potential. The results indicate that fucoidan may have protective action against 6-OHDA-mediated neurotoxicity by modulating oxidative injury and apoptosis through GSH depletion.

Effect of fucoidan on autophagy, migration and invasion of U266 cells / 上海交通大学学报（医学版）

Objective · To investigate effect of fucoidan on autophagy, migration and invasion in human multiple myeloma U266 cells. Methods · The U266 cells treated with fucoidan were cultured in vitro. The formation of autophagosomes was observed by transmission electron microscopy (TEM). Transwell assay was used to evaluate the effect of fucoidan on migratory and invasive abilities of U266 cells. The protein levels of LC3-Ⅱ/LC3-Ⅰ, Beclin-1, P62, MMP9, CXCR4, p-AKT/T-AKT, p-mTOR/T-mTOR were detected by Western blotting. MMP9 concentration in the culture medium was examined by ELISA. Results · ① Autophagosomes increased in fucoidan-treated cells compared with control group under TEM. ② Migratory and invasive abilities were inhibited by fucoidan in a dose-dependent manner, which were suppressed by chloroquine. ③ Western blotting demonstrated that expression of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and MMP9 increased in fucoidan-treated cells, while P62, CXCR4, p-AKT/T-AKT and p-mTOR/T-mTOR decreased compared with control group. ④ The result of ELISA showed that MMP9 concentration in the culture medium of fucoidan-treated cells significantly decreased. Conclusion · Fucoidan induces autophagy and inhibits migration and invasion in U266 cells.

Effects of Fucoidan on Lipidomics Profiling of Juvenile Yellow Catfish (Pelteobagrus fulvidraco) during Feeding Time / 分析化学

The effects of fucoidan on the lipidomics profiling of juvenile yellow catfish (pelteobagrus fulvidraco) during different feeding time (week 1, week 2, week 3 and week 8) were investigated by ultra-performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry (UPLC-Q-TOF-MS) combined with multivariate data analysis, e.g.principal component analysis (PCA) and partial least squares-discriminate analysis (PLS-DA).Based on VIP and p values, 11 lipid biomarkers were screened including lysophosphatidylcholine (Lyso-PC) 16∶0, phosphatidylcholine (PC) 22∶6/16∶0, phosphatidylethanolamine (PE) 22∶6/16∶0, phosphatidylinositol (PI) 18∶0/22∶6, diacylglycerol (DAG) 16∶0/16∶0,DAG 16∶0/18∶1, DAG 18∶1/18∶1, triglycerides (TAG) 20∶5/16∶0/18∶3, TAG 20∶4/16∶1/18∶2, TAG 16∶0/18∶2/20∶5 and TAG 18∶2/14∶0/18∶1.It was found that Lyso-PC, PC, PE and PI had the largest levels at the eighth week;the levels of DAG were decreased at the second and fourth weeks, and increased at the eighth week.While TAG was different: the content of TAG 18∶2/14∶0/18∶1, TAG 20∶5/16∶0/18∶3 and TAG 20∶4/16∶1/18∶2 increased in response to fucoidan, but TAG 16∶0/18∶2/20∶5 decreased little.Therefore, for juvenile yellow catfish, fucoidan can affect its lipid metabolism, which provides a theoretical basis for investigation of the influence of fucoidan on the response mechanism of lipid metabolism of juvenile yellow catfish.

Antioxidant activity of water-soluble polysaccharide extracted from Eucalyptus cultivated in Lebanon

Objective: To extract and identify the chemical composition of the polysaccharide isolated from the Eucalyptus cultivated in Lebanon and to evaluate its antioxidant activity. Materials: The water-soluble polysaccharide was isolated from Eucalyptus leaves, and its structure was identified by Fourier transform infrared spectroscopy, proton nuclear magnetic resonance, and carbon-13 nuclear magnetic resonance. The antioxidant activity of the active ingredient was screened for its radical scavenging ability using 2,2-diphenyl-1-picryl-hydrazyl radical (DPPH) test. Results: The results of the DPPH test have shown that fucoidan, the polysaccharide isolated from Eucalyptus, exhibited almost the same antioxidant activity against DPPH$as the ascorbic acid did at 100 mg/mL. Conclusions: This natural molecule extracted from a medicinal plant has a promising antioxidant activity and could be used in pharmaceutical and medical applications.

Immunostimulatory activity of brown seaweed-derived fucoidans at different molecular weights and purity levels towards white spot syndrome virus (WSSV) in shrimp Litopenaeus vannamei.

Fucoidan used as immunostimulant is commonly in the crude form. In this study, we investigated the effect of fucoidan in both crude and purified forms in their immunostimulatory activity. In addition, we studied the effect of low- and high-molecular weight fucoidan as hydrolysis products toward immunostimulatory activity. Four kinds of fucoidan were assayed for immunostimulant activity on the shrimp Litopenaeus vannamei. The parameters observed in the assay includes the mortality number, haemocyte, gene-related immunity (phenoloxidase, superoxide dismutase and transglutaminase) in the shrimps infected with viral WSSV. The assay results showed that pure fucoidan exhibited higher activity compared with that of crude fucoidan. Sulfate and carbohydrate content of HMW fucoidan are 7.8 % and 82.54 % with an estimated molecular weight of 8.28 x104 Dalton, and low molecular weight (LMW) fucoidan has 1.2% and 65.23% with an estimated molecular weight of 7.53 x104 Dalton. The transcriptional level of the immunity-related genes was found higher after feeding the infected shrimps with purified and HMW fucoidan. In particular, all of fucoidan forms increased the phenoloxidase gene transcription, suggesting that fucoidan have significant role in the production of phenoloxidase.

Fucoidan induces G1 arrest of the cell cycle in EJ human bladder cancer cells through down-regulation of pRB phosphorylation

AbstractFucoidan, a sulfated polysaccharide found in marine algae and brown seaweeds, has been shown to inhibit the in vitro growth of human cancer cells. This study was conducted in cultured human bladder cancer EJ cells to elucidate the possible mechanisms by which fucoidan exerts its anti-proliferative activity, which until now has remained poorly understood. Fucoidan treatment of EJ cells resulted in dose-dependent inhibition of cell growth and induced apoptotic cell death. Flow cytometric analysis revealed that fucoidan led to G1 arrest in cell cycle progression. It was associated with down-regulation of cyclin D1, cyclin E, and cyclin-dependent-kinases (Cdks) in a concentration-dependent manner, without any change in Cdk inhibitors, such as p21 and p27. Furthermore, dephosphorylation of retinoblastoma protein (pRB) by this compound was associated with enhanced binding of pRB with the transcription factors E2F-1 and E2F-4. Overall, our results demonstrate that fucoidan possesses anticancer activity potential against bladder cancer cells by inhibiting pRB phosphorylation.

A comparative study of protective action on endotheliocytes of fucoidan with different molecular weight and its mechanism / 中国药学杂志

OBJECTIVE: To study the protective action on endotheliocytes of fucoidan with different molecular weight from Laminaria japonica, and to explore its mechanism. METHODS: In vivo, endotheliocytes injury model in rats was used to examine the protective action on endotheliocytes of fucoidan with different molecular weight from Laminaria japonica; aortal endothelium section by HE stain under optical microscope was observed, and the the content of von willebrand factor (vWF) in plasma was determined by ELISA. In vitro, endothelial cell culture was used to determine the effect of fucoidan on content of vWF and endothelial microparticles (EMPs). RESULTS: In vivo, aortal endothelium section by HE stain under optical microscope showed low molecular weight (LMW) fucoidan had a remarkable protective effect, and the protection effect of LMWF fucoidans are stronger than middle molecular weight (MMW) fu-coidans. Administration of LMW fucoidan significantly decreased the vWF content. Administration of MMW fucoidan didn't change vWF content. In vitro, Administration of LMW fucoidan significantly decreased the vWF content and EMPs numbers. Administration of MMW fucoidan didn't change the vWF content and EMPs numbers. CONCLUSION: The LMW fucoidan fractions have good protective action on endothelial cells. The protective effect of LMW fucoidans is significantly stronger than MMW fucoidans. The protective action on endotheliocytes of LMW fucoidans relies on decreasing the concentration of EMPs and vWF, and thus inhibiting platelet activation indirectly.

Immunomodulatory effects of sea cucumber fucoidan on macrophage and the signaling pathways / 中国药理学通报

Aim To investigate the immunomodulatory effects of sea cucumber fucoidan ( SC-FUC) on macro-phage and the signaling pathways. Methods Cell via-bilities in response to different concentrations of SC-FUC were analyzed by MTT, phagocytosis ability was detected by neutral red,and nitric oxide ( NO) produc-tion was examined by Griess reaction kit. The mRNA expression levels of IL-6 , IL-10 , Toll-like receptors (TLRs) and related signal molecules MyD88, TRIF, NF-κB were assayed by real-time PCR. All the experi-ments were based on murine RAW264. 7 cell line. Re-sults SC-FUC could promote RAW264 . 7 cell prolif-eration, phagocytosis as evidenced by uptake of neutral red and release of NO. The effects were significant at the early stage (6 h and 12 h) . SC-FUC could up-reg-ulate the expression of IL-6 , IL-10 , TLR4 , TLR5 , TLR9. Moreover, mRNA expressions of TLRs signaling molecules were increased, as well as MyD88, TRIF, NF-κB. Conclusions SC-FUC could activate macro-phage, and then promote the immune function by pro-moting production or expression of NO, IL-6, IL-10. It is speculated to be relevant to activated cell surface re-ceptors in macrophage, including TLR4, TLR5, TLR9, and NF-κB signaling pathways.

Efficacy of Cistanche Tubulosa and Laminaria Japonica Extracts (MK-R7) Supplement in Preventing Patterned Hair Loss and Promoting Scalp Health

Cistanche tubulosa and Laminaria japonica have been reported to have anti-oxidative, anticoagulant, anti-cancer and anti-inflammatory properties. They are expected to be a promising candidates for promoting hair growth and treating dandruff and scalp inflammation as a consequence. In this double-blinded, placebo-controlled clinical trial, we investigated the efficacy of Cistanche tubulosa extract and Laminaria japonica extract complex (MK-R7) in promoting hair health in patients with mild to moderate patterned hair loss. Using phototrichogram (Folliscope 4.0, LeadM, Seoul, Korea), we compared the density and diameter of hairs in patients receiving a placebo or Cistanche tubulosa extract and Laminaria japonica extract complex (MK-R7) at baseline, 8 and 16 weeks of the study. In order to determine the efficacy of treatment on dandruff and scalp inflammation, investigator's assessment score and patient's subjective score were also performed. We found a statistically significant increase in the hair density of the test group (n = 45, MK-R7 400 mg) after 16 weeks of consuming the MK-R7 (test group: 23.29 n/cm2 +/- 24.26, control: 10.35 n/cm2 +/- 20.08, p < 0.05). In addition, we found a statistically significant increase in hair diameter in the test group compared to control group at week 16 (test group: 0.018 mm +/- 0.015, control: 0.003 mm +/- 0.013, p < 0.05). There were also significant outcomes regarding the investigator's visual assessment and patient's subjective score of dandruff and scalp inflammation in the test group compared to those in control group. Based on the results of this clinical study, we conclude that Cistanche tubulosa extract and Laminaria japonica extract complex (MK-R7) are promising substances for promoting health of the scalp and hair.

Antitumor Effects of Fucoidan on Human Colon Cancer Cells via Activation of Akt Signaling

We identified a novel Akt signaling mechanism that mediates fucoidan-induced suppression of human colon cancer cell (HT29) proliferation and anticancer effects. Fucoidan treatment significantly inhibited growth, induced G1-phase-associated upregulation of p21WAF1 expression, and suppressed cyclin and cyclin-dependent kinase expression in HT29 colon cancer cells. Additionally, fucoidan treatment activated the Akt signaling pathway, which was inhibited by treatment with an Akt inhibitor. The inhibition of Akt activation reversed the fucoidan-induced decrease in cell proliferation, the induction of G1-phase-associated p21WAF1 expression, and the reduction in cell cycle regulatory protein expression. Intraperitoneal injection of fucoidan reduced tumor volume; this enhanced antitumor efficacy was associated with induction of apoptosis and decreased angiogenesis. These data suggest that the activation of Akt signaling is involved in the growth inhibition of colon cancer cells treated with fucoidan. Thus, fucoidan may serve as a potential therapeutic agent for colon cancer.