The Active Ingredient Of The The Gynostemma Heat-treated Product Of Anti-NSCLC A549 Cells Research

Heat-processed Gynostemma pentaphyllum by high temperature and high pressure showed more inhibitory activity against non-small cell lung carcinoma (NSCLC) A549cells than that of the crude plant. The objectives of study were to isolate and identify the active constituents and evaluate their A549cell inhibitory activities from the heat-processed G. pentaphyllum. The best conditions of preparation of the heat-processed G. pentaphyllum were confirmed by steam-heating at100,110,120and130℃for1h,2h and3h, respectively. The active fractions were screened using sephadex HP20chromatography and20%,50%,95%ethanol elution. The active constituents were isolated by silica gel, sephadex and reverse phase C18column chromatography, and their chemical structures were identified with NMR, Mass and UV spectra data. The A549cell inhibitory activities of the before and after heat-processing G. pentaphyllum and their active constituents were measured using the3-(4,5-dimethyithiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. The contents of the active constituents of before and after heat-processed G. pentaphyllum were analyzed by UPLC-MS. The results showed that the best condition of preparation of the heat-processed G. Pentaphyllum was stream-heating at130℃,0.24MPa for3h and its ethanol extract appeared more A549cell inhibitory activity than that of the crude G. pentaphyllum. Its IC50value was198.13+7.60μg/mL, whereas for the crude plant it was411.61132.43μg/mL. Using HP20absorption, the95%ethanol elution fraction showed strong A549cell inhibitory activity. From the95%ethanol elution fraction, four dammarane-type saponins were isolated and identified as damulin A, damulin B, gypenoside LI and gypenoside L, respectively. And damulin A and damulin B were found from the heat-processed G. pentaphyllum at fist time. They all appeared strong A549cell inhibitory activities with IC50values of26.98±0.51,4.56±0.58,50.96±9.56and34.94±4.23μg/mL, respectively. Whereas for the positive control ginsenoside Rg3(S) it was39.26±2.05μg/mL. The contents of damulin A, damulin B, gypenoside LI and gypenoside L from the heat-processed G. pentaphyllum preparated at130℃,0.24MPa for3h were11902.19±0.18,5876.64±0.50,2462.52±0.21,6130.26±0.27μg/g, respectively. Heat-processing method was provided for preparation of more active constituents from natural products.