摘要
Objective： To evaluate the effects of the ethanol extract isolated from Weiqi Decoction （胃祺饮, WQD-EE） on AGS cell proliferation and apoptosis. Methods： By using high-performance liquid chromatography with ultraviolet detectors （HPLC-UV） assay and MTT method, the main compounds in WQD-EE and cell viability were detected. And cell cycle distributions were determined by flow cytometry with propidium iodine （PI） staining while apoptosis was detected by flow cytometry with annexin V/PI double staining. Finally, caspase-3 activities were measured by colorimetric method and protein expression was determined by Western blotting. Results： HPLC analysis showed that naringin （35.92 μg/mg）, nobiletin （21.98 μ g/mg）, neohesperidin （17.98μg/mg） and tangeretin （0.756μ g/mg） may be the main compounds in WQD-EE. WQD-EE not only inhibited AGS and MCF 7 cell proliferation in a dose-dependent manner, but also blocked cell cycle progression at G2/M stage as well as inducing cell apoptosis at concentrations triggering significant inhibition of proliferation and cell cycle arrest in AGS cells. While at 0.5 mg/mL, WQD-EE significantly increased caspase-3 activity by 2.75 and 7.47 times at 24 h and 48 h, respectively. Moreover, WQD-EE in one hand reduced protein expressions of p53 and cyclin B1, and in other hand enhanced protein expressions of cytochrome c and Bax. Protein levels of Bcl-2, Fas L and Fas were not significantly affected by WQD-EE. Conclusions： WQD-EE inhibits AGS cell proliferation through G2/M arrest due to down-regulation of cyclin B1 protein expression, and promotes apoptosis by caspase-3 and mitochondria-dependent pathways, but not by p53-dependent pathway.
Objective： To evaluate the effects of the ethanol extract isolated from Weiqi Decoction （胃祺饮, WQD-EE） on AGS cell proliferation and apoptosis. Methods： By using high-performance liquid chromatography with ultraviolet detectors （HPLC-UV） assay and MTT method, the main compounds in WQD-EE and cell viability were detected. And cell cycle distributions were determined by flow cytometry with propidium iodine （PI） staining while apoptosis was detected by flow cytometry with annexin V/PI double staining. Finally, caspase-3 activities were measured by colorimetric method and protein expression was determined by Western blotting. Results： HPLC analysis showed that naringin （35.92 μg/mg）, nobiletin （21.98 μ g/mg）, neohesperidin （17.98μg/mg） and tangeretin （0.756μ g/mg） may be the main compounds in WQD-EE. WQD-EE not only inhibited AGS and MCF 7 cell proliferation in a dose-dependent manner, but also blocked cell cycle progression at G2/M stage as well as inducing cell apoptosis at concentrations triggering significant inhibition of proliferation and cell cycle arrest in AGS cells. While at 0.5 mg/mL, WQD-EE significantly increased caspase-3 activity by 2.75 and 7.47 times at 24 h and 48 h, respectively. Moreover, WQD-EE in one hand reduced protein expressions of p53 and cyclin B1, and in other hand enhanced protein expressions of cytochrome c and Bax. Protein levels of Bcl-2, Fas L and Fas were not significantly affected by WQD-EE. Conclusions： WQD-EE inhibits AGS cell proliferation through G2/M arrest due to down-regulation of cyclin B1 protein expression, and promotes apoptosis by caspase-3 and mitochondria-dependent pathways, but not by p53-dependent pathway.
基金
Supported by High Level Project of University of Educational Commission of Shanghai,Shanghai,China(No.2008GSP19)
Shanghai Municipal Natural Science Foundation,Shanghai China(09ZR1431800)
Opening Project of Shanghai Key Laboratory of Complex Prescription,Shanghai,China(No.11DZ2272300)
Educational Commission of Shanghai,Shanghai,China(No.09JW21 and No.2012JW19)
