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 ultra...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.展开更多
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)+1 种基金Opening Project of Shanghai Key Laboratory of Complex Prescription,Shanghai,China(No.11DZ2272300)Educational Commission of Shanghai,Shanghai,China(No.09JW21 and No.2012JW19)
文摘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.
