Studies On The Active Constituents Of Polygonatum Sibiricum

Objective:For the purpose of identification the active compounds of Ploygonatum sibiricum the constituents of that plant were separated.And the process and screening to active compounds of Ploygonatum sibiricum were optimilized by a novel active induced compounds separation method.And the active tests were used for confirming the results of this method.Methods:1? The constitutes of P.sibiricum were separated,and were identified by NMR.2? In this work,the technology of magnetic beads technology was induced into the research of cell membrane infinite chromatography.The cell membrane magnetic beads “fishing ” active compounds technology was set up for screening the active compounds from traditional Chinese medicine.3? Pharmacology tests were used for evaluating the active of the compounds which were purified.Results:1?The traditional compounds separation was also used for evaluating the constitutes of P.sibiricum.Seventeen compounds were obtained including ten steroidal saponins,six homoisoflavones and one alkaloid.The structures were identified as3-O-?-D-glucopyranosyl(1?4)-?-D-fucopyranosyl-(25R)-spirost-5-en-3?,17?-diol(1),3-O-?-D-glucopyranosyl(1?4)-?-D-fucopyranosyl-(25S)-spirost-5-en-3?,17?-diol(2),3-O-?-D-glucopyranosyl(1?2)-?-D-glucopyranosyl(1?4)-?-D-fuc opyranosyl-(25R)-spirost-5-en-3?,17?-diol(3),3-O-?-D-glucopyranosyl(1?4)-?-D-fucopyranosyl-(25S)-spirost-5-en-3?-ol(4),3-O-?-D-glucopyranosyl(1?4)-?-D-fucopyranosyl-(25R/S)-spirost-5-en-3?,12?-diol(5),3-O-?-D-glucopyranosyl(1?4)-?-D-galactopyranosyl-(25R)-spirost-5-en-3?,17?-diol(6),3-O-?-D-glucopyr anosyl(1?4)-?-D-galactopyranosyl-(25S)-spirost-5-en-3?-ol(7),3-O-?-D-glucop yranosyl(1?2)-?-D-glucopyranosyl(1?4)-?-D-galactopyranosyl-(25R)-spirost-5-en-3?,17?-diol(8),3-O-?-D-glucopyranosyl(1?4)-?-D-galactopyranosyl-(25R/S)-spirost-5-en-3?,12?-diol(9),3-O-?-D-glucopyranosyl(1?4)-?-D-galactopyran osyl-(25R/S)-spirost-5-en-3?-ol-12-one(10),4?,5,7-trihydroxy-6,8-dimethyl-homo isoflavanone(11),4?,5,7-trihydroxy-homoisoflavanone(12),4?,5,7-trihydroxy-6-me thyl-homoisoflavanone(13),2?,5,7-trihydroxy-4?-methoxy-6,8-dimethyl-homoisof lavanone(14),2?,5,7-trihydroxy-4?-methoxy-homoisoflavanone(15),2?,5,7-trihydr oxy-4?-methoxy-8-methyl-homoisoflavanone(16),3-hydroxymethyl-5,6,7,8-tetrah ydroindolizin-8-one(17).Compounds 1 to 5 were new compounds structure.2 ? The macrophage cellular membrane coated magnetic beads(CMMBs)“fishing”-chromatography method was set up.And it was used for screening the anti-inflammation material of P.sibiricum.Induced with the information of this method,the potential active compounds were purified.And the compounds such as 3-O-?-D-glucopyranosyl(1?4)-?-D-fucopyranosyl-(25R)-spirost-5-en-3?,17?-diol and 3-O-?-D-glucopyranosyl(1?4)-?-D-galactosyl-(25R)-spirost-5-en-3?,1 7?-diol were identified by NMR.3?The variation of TNF-? and NO in the supernatant of cells were used as a index to evaluate the anti-inflammatory activity of the purified compounds.And the results were similar with that of CMMBs fishing.4 ? Beside,the glucose consumption was used evaluating the activity of hypoglycemic activity of these compounds.At the level of 10 ?mol/L,compounds 11?12?13 and 16 could reduce the concentration of glucose in the media significantly.It may indicate that these compounds could improve the glucose resistant of 3T3-L1 adipose cell and increase the cells glucose consumption.5 ? The ?-glucosidase inhibition rate was also employed for evaluating the hypoglycemic activity of these compounds.Results showed most of these homoisoflavanones could inhibit ?-glucosidase.And the ?-glucosidase inhibition rate of compounds 14,15 and 16 were 61%?58.1%?64.9%?65.1% respectively.This may indicate these compounds could exert hypoglycemic activity by inhibiting ?-glucosidase.Conclusions:In this work we set up a novel method for the active material of traditional Chinese medicine which is called CMMBs fishing.With the induce of this method the active compounds of P.sibiricum could be purified in a relative short period.Seventeen compounds were separated from P.sibiricum.This work not only could provide a new approach for the study of traditional Chinese medicine but also could provide a basis for the development of P.sibiricum.