Separation Of The Effective Parts Of Honey-processed Austragli And Their Anti-inflammation Activities

Radix Astragali is a commonly used tonic chinese herbal medicine in traditional chinese medicine.The effects of invigorating spleen-stomach and replenishing qi of honey-processed Astragali were significantly improved.Honey-processed Astragali is employed instead of Radix Astragali to achieve improved efficacy in tonifying qi in clinical.This study aimed to separate the effective fractions of honey-processed Astragali and Radix Astragali in high-efficiency and convenient methods,and to observe the in vitro anti-inflammatory effection about effective fractions of honey-processed Astragalus and Astragali.Separating effective parts(polysaccharide,flavonoids,saponins)of honey-processed Astragali and Radix Astragali according to the different polarity.The gel chromatography was used to purify the polysaccharide from honey-processed Astragali and Radix Astragali,study on the molecular weight range of honey-processed Astragali and Radix Astragali polysaccharides by HPLC-ELSD.The monosaccharide composition and content of honey-processed Astragali and Radix Astragali were analyzed by pre column derivatization and UPLC/Q-TOF MS.Poly-sery HLB solid phase extraction method was used to separate and purify the flavonoids and saponins from honey-processed Astragali and Radix Astragali,the chemical composition and relative content of flavonoids and saponins were detected by UPLC/Q-TOF MS combined with area normalization method.Established of LPS-induced mouse RAW264.7 cell inflammatory model,to investigate the cytotoxicity and the regulation of cytokine release about the effective parts of honey-processed Astragali and Radix Astragali.The molecular weight of honey-processed APS was 298847 and the composition of monosaccharides(nmol)was Mannose: Glucose: Xylose: Arabinose: glucuronic acid: Rhamnose=0.06: 28.34: 0.58: 0.24: 0.33: 0.21.While the molecular weight of APS was 309687 and the composition of monosaccharides about APS(nmol)was 0.27: 12.83: 1.63: 0.71: 1.04: 0.56.The relative content of the total flavonoids and Astragaloside of honey-processed Astragalus were 96.05% and 95.06%,including 4 flavonoids and 12 saponins.Combined with our previous experimental research results,the content of total flavonoids was decreased and the content of total saponins was increased after honey-processing.Particularly,the content of Calycosin-7-O-β-D-glucoside-6"-O-malonate was decreased sharply,while the content of calycosin and the total saponins were increased.For effective parts of honey-processed Astragali and Radix Astragali,when concentration was under 300μg/mL,there were no cytotoxic effect on RAW264.7 cells.From the release of inflammatory cytokines,effective parts of honey-processed Astragali and Radix Astragali both have immunosuppressive activity,and the effective parts of honey-processed Astragali were stronger than that of Radix Astragali.By comparing the inhibitory effect on the release of cytokines by the effective parts of honey-processed Astragali,the immunosuppressive activity of saponins was the strongest,followed by the flavonoids and the polysaccharide.While the effective fractions of Radix Astragali had different inhibitory activity on the release of different cytokines.Through studying the monosaccharides composition,we found the content of glucose in honey-processed Astragali polysaccharide was far more than in Radix Astragali polysaccharide,the reason may be honey had a large number of glucose,and the honey-processing made glucose combined with polysaccharides.The content of total saponins was increased while the content of total flavonoids was decreased after honey-processing,may because that honey-processing is beneficial to the dissolution of saponins,the heating process may result in the loss of some perssads and the change of some compounds.The effective parts of honey-processed Astragali could inhibit the release of cytokines NO,IL-1β and TNF-α in inflammatory cells better than the effective parts of Radix Astragali,may be the differences of effective parts of honey-processed Astragali and effective parts of Radix Astragali content and composition result in the difference of immunosuppressive activity.