The Fragments Preparation Of Polysaccharides APS-3b From Angelica Sinensis And Their Anti-tumor Activities

Aim:Angelica sinensis (Oliv), also named as Danggui in China, is a natural plant of Parachute.Its main compounds are ferulic acid, volatile oil and polysaccharides, etc.The previous studies showed that the polysaccharides from Angelica sinensis had anti-tumor,immunoregulation, antiatheroscloresis and antiatheroscloresis biological activities.The water soluble acidic polysaccharide fractions, APS-3b particularly demonstrated significant anti-tumor effect on mice transplanted sarcoma S-180 in vivo and human carcinoma cells in vitro.However, the crude polysaccharides from Angelica sinensis had no obvious inhibitory effect.Therefore, the chemical structures of the polysaccharides from Angelica sinensis seemed to have great relationship with anti-tumor activities.In this research, we used tool enzymes to degrade APS-3b.After purification,different polysaccharide fragments were obtained, and their anti-tumor activities were investigated.These results will help elucidae the relationship between activities and structures of the polysaccharides from Angelica sinensis.Methods:(1) The crude polysaccharide was isolated by methods developed by our laboratory, through anion-exchange and gel chromatography, the acid polysaccharides were obtained.The purified fragments were examined by high performance size exclusion chromatography,polarimeter,phenol-sulfuric acid assay and gas chromatography respectively. (2) According to the structural characteristics of APS-3b, three tool enzymes were used for degradation of them. High performance size exclusion chromatography was used to determined the molecular weights of the purity polysaccharide fragments.(3) Effects of polysaccharides and fragments on the proliferation of three human carcinoma cells, HepG2, HeLa, SW-116 were measured by MTT assay.The models of mice transplanted sarcoma S-180 were used for study the anti- tumor effects in vivo. (4) The monosaccharide compositions of fragments 3b-GAEH-2 and 3b-GAAEH were detected with high performance liquid chromatography.Results:(1)Five polysaccharides, named APS-2b, APS-3b, APS-1b, APS-2c, APS-3c were obtained from Angelica sinensis.All the five polysaccharides were eluted as a single peak in HPSEC, which suggested they were homogeneous polysaccharides. The molecular weights of APS-2b, APS-3b, APS-1b, APS-2c, APS-3c were 7.4×105, 1.1×105, 5.7×105, 2.3×105, 1.4×104.(2) Five polysaccharide fragments were degraded from APS-3b by tool enzymes.The HPSEC analysis indicated that four of the fragments were homogeneous polysaccharides.The molecular weights of 3b-GAEH-1, 3b-GAEH-2, 3b-GLEH, 3b-GAAEH were 3.12×104, 1.29×104, 1.01×105, 8.9×103 Da, respectivively. (3) The effects of five acid polysaccharides and four polysaccharide fragments on the proliferation of three carcinoma cells had great different from each other. 3b-GAEH-2 and 3b-GAAEH showed concomtration-dependent anti-tumor activities, and stornger activities than other acid polysaccharides and fragments.All the acid polysaccharides and fragments inhibited sarcoma S-180 at different degree in vivo.Among them, the average tumor inhibitory of 3b-GAAEH was the hightest and obtained to 49.65% .Furthermore, as two important fragments, 3b-GAEH-2 and 3b-GAAEH showed stronger than acidic polysaccharide APS-2b, APS-3b, APS-1c, APS-2c and APS-3c.(4) The high performance liquid chromatography results demonstrated that the monosaccharide composition of acid polysaccharides were different from each other. The monosaccharide compositions of 3b-GAEH-2 were Ara, Gal, Glc, Rha, in the molar ratio of 8.4:2.7:1.8:1.0. The monosaccharide compositions of 3b-GAAEH were Gal-Rha-Glc-Ara,in the molar ratio of 9.8:2.5:1.2:1.0.Conclusion: The structure of the polysaccharides from Angelica sinensis have great relationship with anti-tumor activities. The acidic polysaccharides from Angelica sinensis all have anti-tumor activities.The fragment 3b-GAEH-2 and 3b-GAAEH mainly composed of arabonose and galactose have most effective anti-tumor activitiesamong all obtained polysaccharides from Angelica sinensis.