Preparation, Purification And Activity Of Polysaccharides Produced By Sargassum Herneri(Tarn)Ag

In the research, SHS (Sargassum herneri polysaccharides) produced by Sargassum.Optimize the traditional extraction methods and their basic properties were analyzed.Theantioxidant activity was evaluated on polysaccharides and the part after purified invitro.Results are as bellow:1. Polysaccharide extraction, purification and the basic character of constructionmaterialsExtrect conditions of SHS produced by Sargassum herneri were optimized accordingto one-factor-at-a-time method and the orthogonal test. Compara the effects of boilingmethod, ultrasonic method and enzymatic method based on the extraction rate. Extractionrates under the optimal solution were: Water extraction4.39%; ultrasonic extraction rate of3.30%; enzymatic5.61%.Three polysaccharides in SHS0, SHS1and SHS0.5were isolatedfrom Sargassum herneri polysaccharides by Q-Sepharose Fast Flow anion-exchangechromatography. The results showed that SHS1monosaccharide composition mainly forMan, Glu, Glc, and the sulfated content of4.48%, protein content of4.71%and the uronicacid content was0.0316μg/8μg. The part of SHS0.5monosaccharide composition mainlyfor Man A, Gul A, sulfated content was2.95%, protein content of5.86%and the uronicacid content was0.072μg/8μg.Polysaccharide sulfated content of4.94%, protein content5.95%, uronic acid content was1.29μg/8μg.2. Study on anti-oxidation activity of SHSStudy on polysaccharide antioxidant activity in vitro. The results indicated thatpolysaccharide has strong antioxidant capacity, and dose-dependent manner.At theconcentration of SHS was10mg/mL, the scavenging rates on DPPH were32.3%; theconcentration of8mg/mL, anti lipid peroxidation rates was48.1%and the concentration of10mg/mL, clearance of superoxide anion was34.2%.Then the protective effect of SHSagainst H2O2-induced RAW264.7murine macrophages oxidative damage was studied.Theresults showed that addition concentration of SHS at200μg/mL after12hours significantlyincreased (P<0.05) the activities of antioxidant enzymes such as SOD and GSH-PX, andsignificantly decreased (P<0.05) the release on intracellular ROS, iNOS, LDH etc. Throughthe RT-PCR experiment, SHS1could significantly enhanced (P<0.05) the expression ofmRNA GSH-Px and Mn-SOD in molecular level. In summary, polysaccharide could protectoxidative damage of RAW264.7cells induced by H2O2through protection of cell structure,.SHS was a type of potent antioxidant.3. Effects of immunoregulation on RAW264.7by SHSThrough the establishment of four models, respectively, the effect of polysaccharidesat cells alone; the effect on cytokines under inflammatory conditions and the function ofprotection and repair on inflammatory cells by SHS. Compared control, LPS and DEX afterSHS treated cells alone, the results showed that SHSCcould significantly increased (P<0.05)the level of TNF-α and IL-10in RAW264.7cells, but it shows the significant differencewith LPS and DEX (P<0.01); At the concentration200μ g/mL of SHS1, compared with thecontrol group and DEX, TNF-α and IL-10levels were significantly increased(P<0.05),besides,compared with LPS (P<0.05). Compared with control addition of SHS0.5alone shows the effect of anti inflammatory (P<0.05), but weaker than DEX. On conditionsof inflammation, low concentration SHS compared with the negative control group. SHS1,has the strongest anti-inflammatory effect than SHSCand SHS0.5. But for highconcentration of SHS,compared with negative control SHSCshows better Antiinflammatory effect than SHS1and SHS0.5. Moreover, compared with SHS, DEX hassignificantly anti-inflammatory (P<0.05).Compared to control SHS0.5could increaseamounts of IL-10showed not significant compared with DEX but significantly with LPS.Furthermore, parallel SHS with control group showed SHS1has more activities primarilyby inhibiting inflammatory factor TNF-α expression play an important role in theprotection and recovery of inflammatory injury cells.group is divided into coarsepolysaccharide comparison of immune besides SHS0.5could decrease the level of IL-10andTNF α compared SHS0.5to LPS. SHS0.5can be adjusted on cellular immune functionIn conclusion, the SHS has stronger antioxidant and immunomodulatoryactivity.Because of SHS is uninfected for cells and easy preparation. This research wasbased on further development of SHS to be enhanceing adjuvants to immunity.