The method of obtaining affects on the composition of polysaccharides Saussurea controversa DC. and their no-stimulating activity

Plant PSs can change the production of nitric oxide by macrophages. To assess the PSs biological activity, independent of contained impurities, purification is required. Purpose. Identify the relationship between the method of obtaining WSPSs from leaves of Saussurea controversa and their ability to stimulate NO-production by mouse peritoneal macrophages. Methods. WSPSs was obtained by extraction with cold (PS1) and hot water (PS2) after pretreatment, extraction with 40% aqueous ethanol (PS3) and with acidified by hydrochloric acid water from raw materials (PS4). The biological activity of WSPSs was studied in C57BL/6 male mice through the NO-production by macrophages using Grace reagent and their proliferation using MTT. The impurity of endotoxin in the samples was determined in a test with antibiotic polymyxin B. Results. The derived WSPSs varied in protein content, uronic acids and monomeric composition. PS2 and PS3 did not affect the viability of macrophages; PS1 and PS4 reduced proliferation relative to intact cells, and PS1 in comparison with LPS. All samples enhanced the nitrites production by macrophages. The suppression of NO-production was detected in cultivation with PS2, PS3 and PS4 treated by polymyxin B. The NO-production during cultivation with PS3, was completely determined by endotoxin. Incubation of polymyxin B with PS1 did not affect its NO-activating properties. Conclusions. Extraction with cold water allows to isolate PSs, free from impurities, with NO-stimulating activity independent of endotoxin, which are promising therapeutic substances for further research of biological activity.

Saussurea controversa DC, polysaccharides, method of production, monomer composition, macrophages, nitric oxide