Purification, Preliminary Characterization, and Immunological Activity of Polysaccharides from Crude Drugs of Sijunzi Formula

Abstract

Sijunzi Decoction (SJZD) is a conventional prescription for curing spleen deficiency in Traditional Chinese Medicine and polysaccharide is its main ingredient. In order to explore the effective ingredients contributing to the immunological activity of SJZD, 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 (GC-3-1) decoctions, respectively. The molecular weight of seven homogeneous polysaccharides ranged from 5.42 × 104 to 5.65 × 104 Da. Monosaccharide composition determined by GC-MS analysis showed that these polysaccharides were primarily composed of Rha, Ara, Xyl, Man, Glc, and Gal with various ratios. Immunological activity assay revealed that polysaccharides from four crude drug components of SJZD displayed inhibitory effects on the complement system. RS-3-1, BZ-3-1, FL-3-1, and GC-3-1 could significantly enhance the phagocytosis and increase the NO production and tumor necrosis factor (TNF-α) level in RAW 264.7 cells (p < 0.05). These results demonstrated the immunological activities of these polysaccharides from the four crude drugs. This study supports the therapeutic effect of SJZD in clinical use and is essential for further identification the immunopolysaccharide from SJZD decoction.

A Comparison of Characterization and Its Actions on Immunocompetent Cells of Polysaccharides from Sijunzi Decoction

“…e immunologic activity of polysaccharides from herbal medicines can be initiated by activating the effector cells such as intestinal lymphocytes of gut immunity [17][18][19], splenic lymphocytes, [20] and macrophages [16,22] of systemic immunity. SJZD is a classic recipe of TCM to strengthen the spleen and replenish Qi [1].…”

“…e eluents whose retention time was 4.8 min with single and symmetrically shark peaks were combined, concentrated, and dried; the residue was collected and named S-3-AG. e homogeneity and molecular weight of Evidence-Based Complementary and Alternative Medicine fraction S-3-AG were evaluated and determined by highperformance gel permeation chromatography (HPGPC) on a TSK-GEL G4000PW × 1 column as described previously [16].…”

“…Characterization Analyses. Monosaccharide composition analyses were determined by GC-MS as previously described [16]. In brief, polysaccharides (5 mg) were hydrolyzed separately with 2 M CF 3 COOH (4 mL) for 6 h at 100°C in a sealed glass tube and concentrated to dry residue by rotary evaporation.…”

“…Our previous study has indicated that S-3, the immunocompetent polysaccharide fraction screened from SJZDP could enhance the immune function of spleendeficiency rats [14] by restoring the disturbance of gut microbiota and increasing the content of short-chain fatty acids. Furthermore, we isolated and purified an immunemodulating polysaccharide (S-3-1) from the S-3 fraction [14,15] and found that the chemical composition of this polysaccharide and sugar residue connection were different from seven homogeneous polysaccharides from four crude drugs (Radix Ginseng, Rhizoma Atractylodis Macrocephalae, Poria, and Radix Glycyrrhizae) of SJZD using the same preparation method [16]. Recently, we isolated and purified a new water-soluble polysaccharide (S-3-AG) from the S-3 fraction; the information on the conformation of S-3-AG is required, and its structure-activity relationships were unclear.…”

“…It is also the active component contributing to the function of intestinal immunoregulation, which can activate immunological response in peyer's patch [24,25], mesenteric lymph nodes [26], intestinal epithelial cells [6], and intestinal intraepithelial lymphocytes [7]. And polysaccharides from four crude drugs of SJZD were demonstrated with macrophage immunomodulatory activities [16]. In order to further explore the immunomodulation activity of SJZDP and its fractions, homogeneous polysaccharide S-3-AG was purified from S-3.…”