Effect of apoptosis on G361 cells by Cimicifuga rhizoma extract / 대한구강보건학회지

OBJECTIVES: To investigate whether the cytotoxic effect of Cimicifuga rhizoma extract is associated with cell death in the human keratinocyte (HaCaT) and human melanoma cell lines (G361). METHODS: Apoptosis induced by Cimicifuga rhizoma extract was confirmed by water-soluble tetrazolium salts-1 (WST-1) assay, immunocytochemistry, and western blot. Additionally, the release of cytochrome c and apoptosis-inducing factor (AIF) was visualized by confocal laser scanning microscopy. RESULTS: The results showed that Cimicifuga rhizoma extract significantly reduced the viability of G361 cells with half-maximal inhibitory concentration (IC 50) of 200 µg/ml, and the apoptotic process was found to occur via the activation of caspase-3 and caspase-9 pathways. Besides, the release of cytochrome c and AIF was also detected. CONCLUSIONS: This study suggests that Cimicifuga rhizoma extract causes apoptosis of human melanoma cells through the intrinsic apoptotic pathway.

HPLC fingerprint of Yinqiao Qingre Tablet / 中草药

Objective: To establish HPLC fingerprint for Yinqiao Qingre Tablet (YQT), and analyze the chemical composition of YQT by ESI-Q-TOF MS. Methods: The Kromasil C18 column (250 mm × 4.6 mm, 5 μm) was used with a mobile phase of acetonitrile-0.1% phosphoric acid in gradient elution, the flow rate was 1.0 mL/min, the column temperature was 25℃, and the detection wavelength was 230 nm. Ultra performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) was used for qualitative analysis in the positive and negative ion modes. Results: The HPLC fingerprint for 11 batches of YQT and 28 common peaks were obtained, belonging to six medicinal herbs. Among them, seven common peaks came from Forsythiae Fructus, nine common peaks came from Lonicerae Japonicae Flos, six common peaks came from Puerariae Lobatae Radix, three common peaks came from Arctii Fructus, one common peak (peak 2) came from Forsythiae Fructus and Cimicifuga Rhizoma, another two common peaks came from Anemarrhenae Rhizoma and Cimicifuga Rhizoma, respectively. The similarity of 11 batches of YQT was over 0.95.Totally 42 chemical components were identified by UPLC-Q-TOF/MS, 16 of which were identified by references, such as puerarin, daidzin, daidzein, neochlorogenic acid, chlorogenic acid, cryptochlorogenin acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, secoxyloganin, rutin, phillyrin, forsythoside A, arctiin, mangiferin, and timosaponin B II, respectively. Conclusion: This study perfects the system of quality standard and provides the reference for the study of substance basis and quality control of the same type of compound preparations.