ABSTRACT
ObjectiveTo investigate the effect of pulsatilla saponin A （PSA） on proliferation and apoptosis of human Burkitt lymphoma （BL） cell line Raji cells and expression of related pathway proteins. MethodWith Raji cells as the research object， the cell proliferation was detected by cell counting kit-8 （CCK-8） method， and the half-maximal inhibitory concentration （IC50） values of 24 h， 48 h and 72 h were calculated to be 19.77， 18.31， 16.70 μmol·L-1， respectively. In subsequent related experiments， 0， 8， 16， 32 μmol·L-1 PSA were selected according to the IC50 value of Raji cells treated with PAS for 72 h. After 0， 8， 16， 32 μmol·L-1 PSA acted on Raji cells for 24， 48， 72 h， the optical density values of cell growth curve were detected by CCK-8 method. The zymogen activities of cysteine aspartate-specific protease （Caspase）-3， Caspase-8 and Caspase-9 in Raji cells treated with 0， 8， 16 and 32 μmol·L-1 PSA for 24 h were measured by Caspase-3， Caspase-8 and Caspase-9 colorimetric assay kit. The apoptosis rate and cell cycle of Raji cells treated with different concentrations of PSA after 24 h were detected by flow cytometry. The expression of B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cleaved poly（ADP-ribose） polymerase （cleaved PARP）， cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3） apoptosis related protein and Janus kinase 2 （JAK2）， signal transducer and activator of transcription 3 （STAT3）， phosphorylated-JAK2 （p-JAK2）， and phosphorylated- STAT3 （p-STAT3） pathway proteins in Raji cells after 24 h of treatment with 0， 8， 16 and 32 μmol·L-1 PSA were tested by Western blot. ResultCompared with control group， decreased cell survival rate， inhibited cell proliferation， activated zymogens of Caspase-3， Caspase-8 and Caspase-9 （P<0.01）， increased apoptosis （P<0.05， P<0.01）， and enhanced cell cycle arrest in Gap phase 2 （G2） were observed in 8， 16 and 32 μmol·L-1 PSA groups（P<0.05， P<0.01）. Compared with control group， cells treated with 8， 16 and 32 μmol·L-1 PSA had lower expression of Bcl-2， p-JAK2， p-STAT3 proteins （P<0.05， P<0.01）， and higher expression of Bax， cleaved PARP and cleaved Caspase-3 protein （P<0.01）， while no significant change was found in the expression of JAK2 and STAT3 proteins. ConclusionPSA could inhibit proliferation and induce apoptosis of Raji cells， and its potential mechanism might be related to the regulation of JAK2/STAT3 signaling pathway.
ABSTRACT
Objective: To investigate the chemical constituents from the berries of Physalis pubescens. Methods: The chemical constituents were isolated by repeated silica gel column chromatography, Sephadex LH-20 gel columns chromatography, medium pressure column chromatography, and semi-preparative liquid chromatography, and their structures were elucidated by chemical properties and spectral analyses. Results: Sixteen compounds were isolated and identified to be 3, 7, 3'-trimethylquercetin (1), kaempferol (2), chrysoeriol (3), quercetin-3-O-β-D-glucopyranoside (4), arjunolic acid (5), pulsatilla saponin A (6), pulsatilla saponin D (7), caffeic acid (8), 1-O-caffeoyl-β-D-glucopyranoside (9), N-trans-ferulogltyramine (10), neo-olivil (11), medioresinol (12), pinoresinol (13), sucrose (14), uridine (15), and β-sitosterol (16). Conclusion: Compounds 5-7 and 9-15 are obtained from the plants of genus Physalis L. for the first time. Compounds 3 and 8 are obtained from this plant for the first time.
ABSTRACT
Objective: To develop an HPLC method for the simultaneous determination of eight saponins in alkali hydrolysate of total saponins from Pulsatilla chinensis. Methods: HPLC was performed on a Kromasil C18 analytical column (250 mm × 4.6 mm, 5.0 μm) at 35°C with MeOH-0.2% HCOOH solution as the mobile phase by gradient elution and the step gradients were as follows: 0-30 min, 70%-100% MeOH; The flow rate was 1.0 mL/min; ELSD gasification chamber temperature was 40°C; Gas pressure of carrier gas N2 was 350 kPa. Results: The linear response (the log values of peak areas with corresponding log values of sample introducing amounts) ranges were 0.799-4.568 μg for pulsatilla saponin D, 0.563-6.756 μg for hederagenin 3-O-β-D-glucopyranosyl-(1→4)-α-L-arabinopyranoside, 0.431-2.683 μg for pulsatilla saponin A, 0.894-7.826 μg for hederacolchiside A1, 0.643-7.504 μg for pulsatilla saponin F, 1.351-7.822 μg for oleanolic acid 3-O-β-D-glucopyranosyl-(1→4)-β-D-glucopyranosyl - (1→3)-α-L-rhamnopyranosyl-(1→2) - α-L-arabinopyranoside, 0.629-2.515 μg for oleanolic acid 3-O-β-D-glucopyranosyl-(1→3)-α-L-rhamnopyranosyl - (1→2)-α-L-arabinopyranoside, and 0.698-2.794 μg for oleanolic acid 3-O-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranoside, respectively (n = 5). The average recoveries of the eight saponins were between 99.0% and 101.0%, and RSD values were less than 1.5%. Conclusion: The results demonstrate that the established method has the adequate accuracy and selectivity for the quality control of alkali hydrolysate of total saponins from P. chinensis.