| Polygonum cuspidatum is the root and rootstock of Polygonum cuspidatum Sieb.et Zucc belonged to Polygonaaceae, which is mainly composed of stilbene and anthraquinones. The reports on α-glycosidase and pancreatic lipase inhibitor concentrated on the water extract of Polygonum cuspidatum which did not report the material basis and the relationship between content and inhibitory activity. It is necessary to to figure out material basis on α- glycosidase and pancreatic lipase of Polygonum cuspidatum to treat diabetes caused by hyperlipidemia.In the study we established the chromatography fingerprint of Polygonum cuspidatum with hyphenated technique of HPLC-DAD-ELSD to select high quality herbs. We selected the comprehensive weighted score of resveratrol, polydatin, emodin and yield as indicators and regarded the amount of solvent, the solvent concentration, the extraction of time and extraction times as the independent variables,optimizing extraction process by using the single factor and response surface methodology. The extracts which extracted by using cyclohexane: ethyl acetate(2:1) and ethyl acetate were from water soluble content of the total extract. The extracts(water, 20% ethanol elution, 40% ethanol and 70% ethanol)which extracted with technique of using macroporous resinwere from water insoluble content of the total extract.We tested the content of stilbene and anthraquinones from different extracts with HPLC and the content of polysaccharide composition with UV, which extracted by extraction solvent and the technique of macroporous resin.We compared pH, temperature and time of enzymatic reaction to optimize the reaction conditions and studied the inhibitory activity of α-glycosidase and pancreatic lipase of different extracts. By testing the content of Fins, GHb, FFA, SOD, MDA, TG, TC, IsI and HOMA-RI, we studied the blood sugar levels of rat with diabetes. The main research results are as follows:The similarities of the chromatography fingerprint with HPLC-DAD-ELSD of Polygonum Cuspidatum from 10 different origin was above 0.9. 19 common peaks were demarcated from the chromatography fingerprint with HPLC-DAD of Polygonum cuspidatum and 14 common peaks were demarcated from the chromatography fingerprint with HPLC-ELSD of Polygonum cuspidatum. The common patterns of the chromatography fingerprint with HPLC-DAD-ELSD was established. Polydatin, resveratrol, emodin-8-O-β-D-glucoside, physcion-8-O-β-Dglucoside, emodin and physcion were identified. In order to select high quality herbs, we compared the peak area from 10 batches of Polygonum cuspidatum.The best extracting procedures by response surface methodology was: extracting them with 15 times amount of 66% ethanol for 2 hours and repeating the process twice.The content of anthraquinones(polydatin, resveratrol, emodin-8-O-β-D-glucoside, physcion-8-O-β-D-glucoside, emodin and physcion) from the cyclohexane extract and ethyl acetate extract was 57.14% and 48.20%. The content of stilbene from the 20% ethanol elution was 13.88%. The content of polysaccharide composition from the water soluble content and water elution was 55.83% and 54.75%.The different extracts from Polygonum cuspidatum had strong inhibitory activity of α-glycosidase and the strength of the inhibitory activity was: water soluble content > water elution > 20% ethanol elution >40% ethanol elution > total extract > part of the rest > water insoluble content > extract from cyclohexane :ethyl acetate(2:1) >70% ethanol elution> ethyl acetate extract.The IC50 of resveratrol was similar to each extract.The inhibitory activity of each extract was better than monomer compounds.Inhibition of type of α-glycosidase of total extract, water soluble content, water elution, 20% ethanol elution, 40% ethanol elution and part of the rest belong to the uncompetitive inhibition. The water insoluble content,70% ethanol elution, extract from cyclohexane: ethyl acetate(2:1) and ethyl acetate extract belong to the competitive inhibition.The different extracts from Polygonum cuspidatum had strong inhibitory activity of pancreatic lipase and the strength of the inhibitory activity was: ethyl acetate extract > extract from cyclohexane:ethyl acetate(2:1) > 70% ethanol elution > water insoluble content > total extract > part of the rest > 40% ethanol elution > 20% ethanol elution > water elution > water soluble content. The ethyl acetate extract was consisting of anthraquinones. The strength of inhibitory activity of monomer compounds was: emodin > physcion> resveratrol > emodin-8-O-β-D-glucoside > physcion-8-O-β-D-glucoside > polydatin. Emodin was best. Inhibition of type of pancreatic lipase of total extract, water insoluble content, extract from cyclohexane: ethyl acetate(2:1),ethyl acetate extract and 70% ethanol elution belong to the competitive inhibition. The part of the rest, water elution, 20% ethanol elution, 40% ethanol elution and water soluble content belong to the uncompetitive inhibition.The total extract, water soluble content and water insoluble content from Polygonum cuspidatum had certain treatment function on the blood sugar levels of rat with diabetes. |
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