RESUMO
Three polysaccharides (SH-1, SH-2 and SH-3) were purified from a brown macroalgea, Sargassum hemiphyllum. The autohydrolysis products from each polysaccharide were separated to three fractions (S fractions as oligomers, L fractions as low molecular weight polysaccharides and H fractions as high molecular weight polysaccharides). Mass spectroscopy of S fractions (SH-1-S, SH-2-S and SH-3-S) showed that these three polymers all contained short stretches of sulfated fucose. The structures of L fractions (SH-1-L, SH-2-L and SH-3-L) were determined by nuclear magnetic resonance (NMR). SH-1-L was composed of two units, unit A (sulfated galactofucan) and unit B (sulfated xylo-glucuronomannan). Unit A contained a backbone of (1, 6-linked ß-D-Gal) n1, (1, 3-linked 4-sulfated α-L-Fuc) n2, (1, 3-linked 2, 4-di-sulfated α-L-Fuc) n3, (1, 4-linked α-L-Fuc) n4 and (1, 3-linked ß-D-Gal) n5, accompanied by some branches, such as sulfated fuco-oligomers, sulfated galacto-oligomers or sulfated galacto-fuco-oligomers. And unit B consisted of alternating 1, 4-linked ß-D-glucuronic acid (GlcA) and 1, 2-linked α-D-mannose (Man) with the Man residues randomly sulfated at C6 or branched with xylose (Xyl) at C3. Both SH-2-L and SH-3-L were composed of unit A and their difference was attributed to the ratio of n1: n2: n3: n4: n5. Based on monosaccharide analysis, we hypothesize that both SH-1-H and SH-2-H contained unit A and unit B while SH-3-H had a structure similar to SH-3-L. An assessment of anti-complement activities showed that the sulfated galactofucan had higher activities than sulfated galacto-fuco-xylo-glucuronomannan. These results suggest that the sulfated galactofucans might be a good candidate for anti-complement drugs.
Assuntos
Fucose/química , Galactose/química , Ácido Glucurônico/química , Polissacarídeos/química , Sargassum/química , Fucose/isolamento & purificação , Galactose/isolamento & purificação , Hidrólise , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Polissacarídeos/isolamento & purificação , Polissacarídeos/ultraestruturaRESUMO
Osteoporosis (OP) is a systemic bone degenerative disease characterized by low bone mass and deteriorated microarchitecture of bone tissue, causing high morbidity and mortality rates. Bone resorption by overactivated osteoclasts (OCs) is the main cause of osteoporosis. Glucuronomannan and its oligomers (Gs) and their sulfated derivatives (SGs) were previously prepared. The anti-osteoporosis activities of these glycans were evaluated. Firstly, we determined the viability of RAW264.7 by CCK-8 test. Nextly, we investigated the inhibitory effects of Gs and SGs on the differentiation of RAW264.7 cells into OCs using tartrate-resistant acid phosphatase (TRAP) staining, F-actin ring staining, qualitative reverse-transcription polymerase chain reaction(qRT-PCR) and western blotting. TRAP staining revealed that Gs significantly blocked RANKL-induced OC generation while SGs did not exhibit this ability. F-actin staining assays demonstrated that Gs inhibits RANKL-induced actin ring formation. qRT-PCR analyses indicated that Gs dose-dependently inhibited the expression of OCs marker genes including Trap, NFATc1, c-Fos, DC-Stamp and ATP60 during the differentiation process, while SGs did not suppress. Regarding the mechanism of Gs, it was found that Gs suppressed osteoclastogenesis via inhibiting the degradation of IRF-8 and interfering with NF-κB pathway activation. Together, these results suggest that Gs have the ability to inhibit osteoclastogenesis by modulating IRF-8 signaling.
Assuntos
Osteoporose , Sargassum , Actinas , Diferenciação Celular , NF-kappa B/metabolismo , Fatores de Transcrição NFATC , Oligossacarídeos/farmacologia , Osteoclastos , Osteogênese , Osteoporose/metabolismo , Sargassum/metabolismo , Animais , CamundongosRESUMO
Nonalcoholic fatty liver disease (NAFLD) is considered as the hepatic manifestation of metabolic syndrome, ranging from benign steatosis to severe non-alcoholic steatohepatitis. Recently, it has been found that lipophagy plays a pivotal role in lipid turnover, which can alleviate NAFLD in hepatocytes. In this study, we found that a highly sulfated glucuronomannan hexamer G6S1 has the ability to enhance lipophagy. When treated with G6S1, the number and the size of lipid droplet (LD) decreased significantly on hepatocytes AML12 cells. Western blot results showed that the expressions of the lipolysis-related proteins increased, while the expressions of proteins that is responsible for lipid transportation and synthesis exhibited no significant change. Immunofluorescence assay and electron microscopy results showed an increase of autophagy related protein expression level and lysosome number in hepatocytes treated with G6S1, suggesting that G6S1 could also promote lipophagy. A significant increase of peroxisome proliferator-activated receptor alpha (PPARα) expression level was detected in G6S1 treated cells, suggesting that G6S1 may promote autophagy via enhancing the expression of PPARα. In addition, these effects could be inhibited after treatment with autophagy inhibitor 3-methyladenine (3-MA) and PPARα inhibitor MK-886. These findings indicate that G6S1 can promote lipophagy via enhanced PPARα expression and can result in a slowdown of lipids accumulation.
Assuntos
Lipólise , PPAR alfa , Humanos , Hepatopatia Gordurosa não AlcoólicaRESUMO
A sulfated glucurono-xylo-rhamnan (EP-3-H) was purified from a green alga, Enteromorpha prolifera. EP-3-H and its oligomers were characterized by high performance liquid chromatography, mass spectrometry and one and two-dimensional nuclear magnetic resource spectroscopy. The structural analysis showed EP-3-H has a backbone of glucurono-xylo-rhamnan, branches with glucuronic acid and sulfated at C3 of rhamnose and/or C2 of xylose. The inhibition of EP-3-H on human lung cancer A549 cell proliferation in vitro and its therapeutic effects in BALB/c-nu mice in vivo were determined to evaluate the anti-lung cancer activity of EP-3-H. The tumor inhibition level was 59 %, suggesting that EP-3-H might be a good candidate for the treatment of lung cancer. Surface plasmon resonance (SPR) studies revealed the IC50 on the binding of fibroblast growth factors, (FGF1 and FGF2), to heparin were 0.85 and 1.47 mg/mL, respectively. These results suggest that EP-3-H inhibits cancer proliferation by interacting with these growth factors.
Assuntos
Antineoplásicos , Desoxiaçúcares , Neoplasias Pulmonares/tratamento farmacológico , Mananas , Células A549 , Animais , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Proliferação de Células/efeitos dos fármacos , Desoxiaçúcares/isolamento & purificação , Desoxiaçúcares/farmacologia , Desoxiaçúcares/uso terapêutico , Fator 1 de Crescimento de Fibroblastos/metabolismo , Fator 2 de Crescimento de Fibroblastos/metabolismo , Heparina/metabolismo , Humanos , Neoplasias Pulmonares/metabolismo , Masculino , Mananas/isolamento & purificação , Mananas/farmacologia , Mananas/uso terapêutico , Camundongos Endogâmicos BALB C , Camundongos Nus , Sulfatos , Ulva/químicaRESUMO
The anti-lung cancer activity of oligosaccharides derived from glucuronomannan was investigated. The inhibition of A549 cell proliferation by glucuronomannan (Gn) and its oligomers (dimer (G2), tetramer (G4) and hexamer (G6)) were concentration dependent. In vivo activities on the A549-derived tumor xenografts showed the tumor inhibition of G2, G4 and G6 were 17 %, 40 % and 46 %, respectively. Organ coefficients in nude mice showed an increase in the kidney with G4, the brain with G6, and the spleen with G6. An advanced tandem mass tag labeled proteomics approach was performed. A significant differential expression was found in 59 out of the 4371 proteins, which involved the immune system. Surface plasmon resonance (SPR) studies revealed G6 was strongly bound to immunoglobulin G. This suggests that glucuronomannan hexamer inhibits the proliferation of lung cancer through its binding to immunoglobulin.