Study On Tracing The Source Of The Immune Active Polysaccharide From Sijunzi Decoction

Sijunzi Decoction(SJZD)is a conventional prescription for curing spleen deficiency in Traditional Chinese Medicine,and consists of four crude drugs: Radix Ginseng,Rhizoma Atractylodis Macrocephalae,Poria,and Radix Glycyrrhizae.In our previous study,we screened one immune polysaccharide from SJZD decoction.In order to explore the effective ingredients contributing to the immunological activity of SJZD in the present study,we isolated and purified seven homogeneous polysaccharides from Radix Ginseng(RS-3-1 and RS-3-2),Rhizoma Atractylodis Macrocephalae(BZ-3-1,BZ-3-2 and BZ-3-3),Poria(FL-3-1),and Radix Glycyrrhizae decoctions(GC-3-1),respectively,the molecular weight of which ranged from 5.42×104 to 5.65×104 Da.Monosaccharide composition determined by GC-MS analysis showed that these polysaccharides were primarily consisted of Rha,Ara,Xyl,Man,Glc and Gal with various ratios.Immunological activity assay revealed BZ-3-1 displayed a significant inhibitory effect on the complement system,with CH50 values of 0.482 mg/mL.Moreover,RS-3-1,BZ-3-1,FL-3-1,GC-3-1 could enhance RAW 264.7 cells phagocytosis,promote NO production,improve tumor necrosis factor(TNF-α)levels(p < 0.05).Structural characterization of active polysaccharides RS-3-1,BZ-3-1,FL-3-1 and GC-1 were investigated by IR,Methylation,Periodate oxidation,Smith degradation and NMR spectrum.Compared the the molecular weight,monosaccharide composition and ratio of homogeneous polysaccharides from four crud drugs of this prescription,Radix Ginseng,Rhizoma Atractylodis Macrocephalae,Poria,and Radix Glycyrrhizae with immunopolysaccharide S-3-1 from the SJZD,there are no exactly same polysaccharide.It is suggested that the active compound S-3-1 may be from the decocting process.In conclusion,this study investigated the polysaccharides isolated from four crud drugs which comprised of SJZD,which could help to track the the origin of SJZD immunopolysaccharide S-3-1 and provide a support for clinical therapeutic effect of SJZD.However,the material basis of SZJD immune activity is still not clear and comprehensive researches for the immune mechanism and structure–activity relationship are wanted.