Pine Pollen Polysaccharide Extraction, Purification, Sulfation And Activity Research

A water soluble crude polysaccharide was extracted with hot water andprecipitated by alcohol from Pinus massoniana pollen. Protein was removedby Sevag method. The pollen polysaccharides of pine were white powder andthe efficiency of extraction was 3.256±0.230%. Four fractions were obtainedrespectively by the method of ethanol grading and the 60% polysaccharidewas chemically modified by chlorosulfonic axid-pyridine method.No typical absorption of nucleic acid or protein was observed in UVspectrum; The feature bands of pollen polysaccharides and the S-60% pollenpolysaccharide of pine were demonstrated in the special infrared spectrumregions. The 60% pollen polysaccharide had 83.24% saccharide and theS-60% pollen polysaccharide, a modified compound with 1.471 substituteddegrees was obtained. We also tested the composition of the pollenpolysaccharide: the 40% polysaccharide mainly contained glucose, arabinoseand galactose and a small ratio of mannose, glucuronic acid and fucose; the60% polysaccharide mainly contained arabinose and galactose and a smallratio of glucose; the 80% polysaccharide contained galactose, glucuronic acid,mannose and glucose and a small ratio of rhamnose and fucose.Then the 40%polysaccharide was further purified with column chromatography onSepharose CL-6B and the NMR Spectrum showed that the composition ofpurified polysaccharide also was two or more monosaccharide.We tested the anticancer activities of the polysaccharides from pine pollen by three kinds of cancer cell: BGC-823,HepG2,A549 in vitro an S180tumour in vivo. Results showed that the S-60% polysaccharide could inhibitthe growth of all the three kinds of cancer cell in vitro an S180 tumour in vivoremarkably. In vitro, the inhibitory ratio of HepG2 was the highest, to 39.90%.In vivo, The highest inhibitory ratio of S180 tumour was 69.27%.The results of immunocyte research showed that the polysaccharidesfrom pine pollen could promote immunocyte proliferation remarkably. Thehighest promoted ratios of B, T-lymphocyte proliferation and macrophageactivation were 221.4%, 155.3%, and 53.44%.We first discovered that the S-60% polysaccharide could inhibit thecontractilities of small intestines of mouse. The 60% polysaccharide andS-60% polysaccharide could mainly promote heart contractilityies and inhibitheart rate of Chinese toads, studied by heart perfusion.