Qingguo Han

Elucidation of the Molecular-Mechanisms and In Vivo Evaluation of the Anti-inflammatory Effect of Alginate-Derived Seleno-polymannuronate

Alginate-derived polymannuronate (PM) is a type of polysaccharide found in edible brown seaweeds. Seleno-polymannuronate (Se-PM) was prepared from PM via synthesis using sulfation- and selenation-replacement reactions. The anti-inflammatory activity of Se-PM and its corresponding molecular mechanisms were investigated. In lipopolysaccharide (LPS)-activated murine macrophage RAW264.7 cells, Se-PM significantly attenuated the production of nitric oxide (NO), prostaglandin E2 (PGE2), and reactive oxygen species (ROS); the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2); and the secretion of proinflammatory cytokines. Moreover, Se-PM remarkably suppressed the LPS-induced activation of the nuclear-factor (NF)-κB and mitogen-activated-protein-kinase (MAPK) signaling pathways in RAW264.7 cells. Furthermore, Se-PM also decreased the production of proinflammatory mediators in LPS-triggered primary murine macrophages. Additionally, Se-PM inhibited the inflammatory response in the air-pouch inflammation model. These results might contribute to the overall understanding of the potential health benefits of Se-PM for food and drug applications.

Assessment of pollutions and identification of sources of heavy metals in sediments from west coast of Shenzhen, China

The sediment samples were collected from eight sites located in the Pearl River Estuary and the Shenzhen Bay of the west coast of Shenzhen. The distributions of the seven elements Zn, Cr, Hg, Cu, Cd, Pb and As have been analyzed, and their pollution degrees, corresponding potential ecological risks and source identifications have been studied using geo-accumulation index, potential ecological risk index and integrated multivariate statistical methods, respectively. Based on the calculated geo-accumulation indices, the contamination levels of all elements in the Pearl River Estuary are similar to those in the Shenzhen Bay, reflecting that these elements in the study areas have similar sources because of the adequate seawater exchange. The calculated potential ecological risk indices suggest that Cd and Hg are at considerable and moderate risk, respectively. Multivariate statistical analyses further reveal that Zn, Hg, Cd and Pb originated from industrial wastewater, while Cr and Cu are mainly from both industrial wastewater and agricultural sources, and As is mainly from natural source. These research results provide baseline information for both the coastal environment management and the worldwide heavy metal distribution and assessment.

Exploring inhibition mechanism and nature of lipase by Ligupurpuroside A extracted from Ku-Ding tea

Enzyme-activity essay demonstrated that Ligupurpuroside A acts as a natural inhibitor of lipase in a competitive manner. The inhibition mechanism and nature of lipase by Ligupurpuroside A were investigated by fluorescence spectra, UV–Vis absorption spectra, circular dichroism (CD) spectra and molecular docking methods. Fluorescence experiments indicated that Ligupurpuroside A can quench the intrinsic fluorescence of lipase through a static quenching procedure. Thermodynamic analysis suggested that hydrophobic interaction is the main force between lipase and Ligupurpuroside A. Fluorescence resonance energy transfer experiment showed that an energy transfer from lipase to Ligupurpuroside A occurs with great possibility, confirming the presence of static quenching mechanism of lipase fluorescence by Ligupurpuroside A. Interestingly, conformation of lipase remained almost the same after bound to Ligupurpuroside A based on CD spectral experiments. All these experimental results were validated by the protein-ligand docking studies which further showed that Ligupurpuroside A could bind to the amino acid residues of the hydrophobic cavity on catalytic sites of lipase. This study should be helpful for the design of diet drug and the full use of Ligupurpuroside A in food industry.

Alginate enhances Toll-like receptor 4-mediated phagocytosis by murine RAW264.7 macrophages

Alginate is a naturally acidic polysaccharide consisting alternately of β-d-mannuronic acid and α-l-guluronic acid with 1, 4-glycosidic linkages and is derived from brown seaweeds. Herein, the effect of alginate on the promotion of macrophage phagocytosis and the corresponding molecular mechanisms were investigated in murine RAW264.7 cells. Alginate could enhance the intracellular phagocytosis of gold nanoparticles (AuNPs), fluorescent microspheres and immunoglobulin G (IgG)-opsonized Staphylococcus aureus (S. aureus). Moreover, alginate increased Toll-like receptor 4 (TLR4) expression and activated the Akt/nuclear factor-κB (NF-κB) and p38 mitogen-activated protein kinase (MAPK) signalling pathways. Alginate-promoted phagocytosis was suppressed by the addition of inhibitors of TLR4, NF-κB and p38 MAPK and by TLR4 gene knockdown, indicating the involvement of these key components. This work is the first to propose that alginate promotes phagocytosis via upregulating TLR4 expression and stimulating the Akt/NF-κB and p38 MAPK signalling pathways, which may contribute to the capacity of alginate to activate macrophages.

Trypsin inhibition by Ligupurpuroside B as studied using spectroscopic, CD and molecular docking techniques

Abstract It is well known that Ligupurpuroside B is a water-soluble polyphenolic compound and used to brew bitter tea with antioxidant activities. It acted as a stimulant to the central nervous system and a diuretic (increase the excretion of urine), was used to treat painful throat and high blood pressure, and also exerted weight-loss function. In this regard, a detailed investigation on the mechanism of interaction between Ligupurpuroside B and trypsin could be of great interest to know the pharmacokinetic behavior of Ligupurpuroside B and for the design of new analogues with effective pharmacological properties. Ligupurpuroside B successfully quenched the intrinsic fluorescence of trypsin via static quenching mechanism. The binding constants (Ka) at three temperatures (288, 298, and 308 K) were 1.7841 × 104, 1.6251 × 104 and 1.5483 × 104 L mol−1, respectively. Binding constants revealed the stronger binding interaction between Ligupurpuroside B and trypsin. The number of binding sites approximated to one, indicating a single class of binding for Ligupurpuroside B in trypsin. The enzyme activity result suggested that Ligupurpuroside B can inhibit trypsin activity. Thermodynamic results revealed that both hydrogen bonds and hydrophobic interactions play main roles in stabilization of Ligupurpuroside B-trypsin complex. Circular dichroism (CD) results showed that the conformation of trypsin changed after bound to ligupurpuroside B. Molecular docking indicated that Ligupurpuroside B can enter the hydrophobic cavity of trypsin and was located near Trp215 and Tyr228 of trypsin. Communicated by Ramaswamy H. Sarma

Immune activation of RAW264.7 macrophages by low molecular weight fucoidan extracted from New Zealand Undaria pinnatifida

Fucoidan, a sulfated polysaccharide extracted from brown seaweeds, has been shown to possess various bioactivities. In particular, low molecular weight fucoidan (LMWF) has been shown to have better bioactivities. In this study, a LMWF (<10 kDa) was extracted from New Zealand Undaria pinnatifida and investigated for its immune modulation effects. LMWF at a concentration range from 1 to 50 μg/mL exerted an effective immune activation in RAW264.7 macrophages. LMWF treatment promoted significant NO release, iNOS expression, and TNF-α and IL-6 secretion in a concentration-dependent manner. It also significantly stimulated the activation of NF-κB and MAPK signaling pathways, and specific inhibitors of NF-κB and MAPK pathways diminished the stimulation, confirming the activation pathways. These results indicate that LMWF possesses potential health benefits through immune-stimulation, which may lead to future pharmaceutical development.

Binding mechanism of lipase to Ligupurpuroside B extracted from Ku-Ding tea as studied by multi-spectroscopic and molecular docking methods

The interaction of lipase with Ligupurpuroside B was studied by multiple spectroscopic techniques, enzyme activity and molecular modeling under simulative physiological condition. According to Stern-Volmer equation, fluorescence of lipase was quenched by Ligupurpuroside B via a static quenching mechanism because of formation of Ligupurpuroside B-lipase complex. Binding constants, number of binding sites & thermodynamic parameters were evaluated. The values of ΔGo (-25.085 kJ mol-1), ΔHo (-12.14 kJ mol-1) and ΔSo (+43.45 J mol-1 K-1) at 298 K indicated that Ligupurpuroside B-lipase interaction is spontaneous and hydrophobic interaction is the main force stabilizing the Ligupurpuroside B-lipase complex. The enzyme activity assay showed that Ligupurpuroside B inhibited lipase activity efficiently. Synchronous fluorescence spectra (SFS) suggested that Ligupurpuroside B is closer to Trp residues than to Tyr residues. All above experimental results were confirmed by molecular docking studies, which further indicated the binding site of Ligupurpuroside B on the surface of lipase, and the amino acid residues of lipase interacting with Ligupurpuroside B. Our present research work gives valuable information on the design of drugs with lipase as a carrier and should be useful for food industries.