Immunoadjuvent activity of different polysaccharides from Sijunzi decoction and its components

  Objective  To study the effect of water-soluble polysaccharides from Sijunzi decoction and its four components on mouse macrophage cells.

  Methods  The crude polysaccharides from Sijunzi decoction (composed of ginseng, atractylodes, tuckahoe and Radix glycyrrhizae preparata) were extracted with aqueous extraction and alcohol precipitation and then separated into 3 groups of high (> 100 kD), medium (30 to 100 kD), and low (< 30 kD) molecular weight using centrifuge through filter membrane. The same procedures were adopted to acquire crude polysaccharides form the four components of Sijunzi decoction. The mouse macrophage cells RAW264.7 were treated with the extracted polysaccharides at the dosage of 100 mg/L for 24 hours; then the cell cycle, phagocytosis, and reactive oxygen species (ROS) of the cells were determined with flow cytometry (FCM). The expressions of CD40, CD86 and CD284 were also detected with FCM to analyze polarizations of the RAW264.7 cells.

  Results  The low and medium molecular weight polysaccharides from Sijunzi decoction and its two components (ginseng and Radix glycyrrhizae preparata) promoted the proliferation and increased the proportions of the cells in S and G2M of the RAW264.7 cells. The ratios of CD40 positive cells were 86.30% ± 3.72%, 87.19% ± 2.76%, and 85.48% ± 4.63% for the RAW264.7 cells treated with medium molecular weight polysaccharides from Sijunzi decoction, ginseng and Radix glycyrrhizae preparata, and all obviously higher than that (76.99% ± 3.28%) of control cells. Compared to those of the control cells, the RAW264.7 cells treated with medium molecular weight polysaccharides from Sijunzi decoction, ginseng and Radix glycyrrhizae preparata also showed higher ratios of CD86 positive (65.60 ± 2.35%, 65.72 ± 2.35%, and 65.72 ± 2.70% vs. 60.64 ± 2.25%) and CD284 positive (7.46 ± 0.46, 7.22 ± 0.14, and 7.54 ± 0.28 vs. 6.24 ± 0.74).

  Conclusion  The medium molecular weight polysaccharides from Sijunzi decoction, ginseng and Radix glycyrrhizae preparata could upregulate expressions of CD40, CD86 and CD284 and promote the polarizations of RAW264.7 cells; the low and medium molecular weight polysaccharides from Sijunzi decoction and its components have strong immunoadjuvent activity.