Investigation of natural deep eutectic solvent for the extraction of crude polysaccharide from Polygonatum kingianum and influence of metal elements on its immunomodulatory effects

In this study, natural deep eutectic solvent (NADES) was used to extract Polygonatum kingianum crude polysaccharide (PKCP) and response surface methodology (RSM) was designed to optimize the extraction procedure. The immunomodulatory effect of PKCP and the influence of metal elements on its immunomodulatory effect were further discussed. The optimum conditions for PKCP extraction were obtained by RSM optimization: NADES were synthesized with a 1:2 choline chloride -glycerol molar ratio, then extracted at a liquid -solid ratio of 16.6 mL g(-1) and water content of 31.2 % for 60 min at 60 C-degrees. This method was used for the extraction of PKCP, and the extraction efficiency was 29.69 %, which was 2.5 times greater than the conventional method of water extraction. In the concentration range of 200-800 mu g mL(-1), PKCP could activate macrophages, promoting NO secretion and mRNA expression of interleukin-6 (IL -6) and inducible nitric oxide synthase (iNOS) in a dosedependent way. NO secretion and cytokine expression were not affected when the metal elements were spiked to the equivalent of the metal elements contained in Polygonatum kingianum. When the content of metal elements was higher, the secretion of NO and the gene expression of iNOS were both decreased, which may affect the immunomodulatory effect of Polygonatum kingianum.