The normal cell proliferation and wound healing effect of polysaccharides from Ganoderma amboinense

To study the cell proliferation and wound healing activity of polysaccharides from Ganoderma amboinense(GAMPS),the polysaccharide was extracted by water extraction and alcohol precipitation method,and its monosaccharide composition and molecular weight were analyzed.The effects of different concentrations of GAMPS on the cell proliferation were determined by cell survival rate test,and the wound healing ability of GAMPS to NIH/3T3 cells was detected.The preliminary evaluation of the antioxidant ability of GAMPS was conducted by the oxygen radical absorbance capacity(ORAC).The results showed that the GAMPS was composed of glucose,mannose,and galactose at a molar ratio of 67.62:14.07:7.50,and the weightaverage molecular weights were 5.439×10^(6) and 1.704×10^(5) g/mol by using high-performance gel-permeation chromatography-multiple angle laser scatter(HPGPC-MALS)analysis.GAMPS(0.2μg/μL)showed the strongest proliferation ability to THP-1 cells,with cell survival rate of 178.7%.The wound healing effect of GAMPS(0.1μg/μL)was obvious on NIH/3T3 and 3.75μg/μL of GAMPS showed the strongest total antioxidant ability.All the results indicate that GAMPS promotes cell proliferation,and has cell wound healing effect and strong antioxidant activity.The results provide theoretical foundation for the development and utilization of GAMPS.

Comparison of Structure and Biological Activity of Natural Polypeptide from Velvet Antlers of Cervus elaphus with Those of Synthesized Polypeptide

Biochemical techniques including ion-exchange chromatography, gel filtration chromatography and re- versed-phase high-performance liquid chromatography（RP-HPLC） were used to isolate and purify the natural poly- peptide from velvet antler（nVAP） of Cervus elaphus（C, elaphus）, which has a molecular weight of 3215.8 and the primary structure of VLSAADKSNVKAAWGKVGGNAPAFGAEALLRM. The homology of the protein sequence in nVAP with known protein sequence is less than 50%, suggesting that nVAP appears to be a new bioactive substance. At a level of 0.4--50 gg/mL, nVAP promotes mitosis in epidermal ceils, chondrocytes and NIH3T3 fibroblasts pri- marily cultured in a significant way. Given that a yield of high-purity nVAP isolated from C. elaphus is 0.001%, nVAP is artificially synthesized to prepare synthetic velvet antler polypeptide（sVAP） according to its primary struc- ture. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE） of sVAP shows a single band, and its HPLC spectrum displays a single peak. The matrix-assisted laser desorption ionization time-of-flight mass spectro- metry（MALDI-TOF-MS） was used to identify sVAP to be of a molecular weight of 3200 and the consistency between primary structures of sVAP and nVAE Bioactivity test shows that at a dose of 5--40 μg/mL, sVAP promotes the pro- liferation of primarily cultured epidermal cells and NIH3T3 cell line. From the traditional Chinese medicine theory, velvet antler from Cervus nippon（C, nippon） and velvet antler from C. elaphus are considered as the same medicine, but differences between biochemical base and pharmacological effect of these two velvet antlers have been observed. We compared the total polypeptide mapping of the two velvet antlers, discovering that nVAP is active polypeptide and only exists in the velvet antler of C. elaphus, sVAP is similar to nVAP in physicochemical property and biological activity. These studies extend the possible utility of sVAP to be the promising compound to prepare velvet antler polypeptide of C. elaphus.