The anti-tumor effects of catechins and gallic acid （GA） in-vitro was investigated in this paper. Fluo-3AM, Calcium-AM （Ca-AM）, 2＇, 7＇ -dichlorofluorescein-diacetate（DCFH-DA）, 4＇ ,6-dia- midino-2- phenylindo...The anti-tumor effects of catechins and gallic acid （GA） in-vitro was investigated in this paper. Fluo-3AM, Calcium-AM （Ca-AM）, 2＇, 7＇ -dichlorofluorescein-diacetate（DCFH-DA）, 4＇ ,6-dia- midino-2- phenylindole （DAPI） and Ca-AM plus colbat were used to characterize intracellular calcium, labile iron pool （LIP）, reac- tive oxygen species （ROS）, nuclei morphology and mitochondrial permeability transition pore （mPTP） opening, respectively. High performance liquid chromatography （HPLC） was used to quanti- tare catechins and GA in the cultural medium. The results indi- cated that each of them showed dose response inhibition of cell growth, provoking nuclei condensation, intracellular calcium ele- vation, mPTP opening, LIP reduction, and cytochrome c （Cyt-C） to release into cytosol. The caspase inhibitors, 2-aminoethox- ydiphenol borate （APB） or Fe3＋ could inhibit lethal effects of GA and （-）-epigallocatechin （EGC）, but failed to affect （-）-epigalloca- techin gallate （EGCG） and （-）-epicatechin gallate （ECG）. Level of ROS presented negative growth while their concentration de- creased in the medium. In conclusion, our findings suggest that viability of RKO decreased because of their good correlation with elevation of calcium and loss of L1P and ROS in cytosol.展开更多
Divicine is an active pyrimidine aglycone,generated from vicine by the enzymeβ-glucosidase upon ingestion of fava beans.In this study,we investigated the effect of divicine on cultured human umbilical vein endothelia...Divicine is an active pyrimidine aglycone,generated from vicine by the enzymeβ-glucosidase upon ingestion of fava beans.In this study,we investigated the effect of divicine on cultured human umbilical vein endothelial cells(HUVECs)and explored the potential mechanisms.Incubation HUVECs with 18.5–85.1μM divicine resulted in a concentration and time dependent decrease of cell viability,followed by decrease of cellular reduced glutathione,as well as increase of reactive oxygen species(ROS),malondialdehyde(MDA),and labile iron pool.Transmission electron microscopy confirmed that the divicine treated HUVECs’mitochondria had shrunk.Importantly,the administration of desferrioxamine,an iron chelator,to the divicine treated HUVECs significantly reduced iron overload and cell death and decreased cellular ROS and MDA.These results demonstrated that divicine could cause damaging of endothelial cells,and ferroptosis might be involved in divicine induced HUVECs injury,reminding long term ingestion of fava beans might be harmful to vascular system,especially for those suffering from glucose 6-phosphate dehydrogenase deficiency.展开更多
基金Supported by Zhejiang Provincial Natural Science Foundation of China(Y3100683)
文摘The anti-tumor effects of catechins and gallic acid （GA） in-vitro was investigated in this paper. Fluo-3AM, Calcium-AM （Ca-AM）, 2＇, 7＇ -dichlorofluorescein-diacetate（DCFH-DA）, 4＇ ,6-dia- midino-2- phenylindole （DAPI） and Ca-AM plus colbat were used to characterize intracellular calcium, labile iron pool （LIP）, reac- tive oxygen species （ROS）, nuclei morphology and mitochondrial permeability transition pore （mPTP） opening, respectively. High performance liquid chromatography （HPLC） was used to quanti- tare catechins and GA in the cultural medium. The results indi- cated that each of them showed dose response inhibition of cell growth, provoking nuclei condensation, intracellular calcium ele- vation, mPTP opening, LIP reduction, and cytochrome c （Cyt-C） to release into cytosol. The caspase inhibitors, 2-aminoethox- ydiphenol borate （APB） or Fe3＋ could inhibit lethal effects of GA and （-）-epigallocatechin （EGC）, but failed to affect （-）-epigalloca- techin gallate （EGCG） and （-）-epicatechin gallate （ECG）. Level of ROS presented negative growth while their concentration de- creased in the medium. In conclusion, our findings suggest that viability of RKO decreased because of their good correlation with elevation of calcium and loss of L1P and ROS in cytosol.
基金This work was supported by the National Natural Science Foundation of China(31671183)the Natural Science Foundation of Guangdong Province(2018A030307005)。
文摘Divicine is an active pyrimidine aglycone,generated from vicine by the enzymeβ-glucosidase upon ingestion of fava beans.In this study,we investigated the effect of divicine on cultured human umbilical vein endothelial cells(HUVECs)and explored the potential mechanisms.Incubation HUVECs with 18.5–85.1μM divicine resulted in a concentration and time dependent decrease of cell viability,followed by decrease of cellular reduced glutathione,as well as increase of reactive oxygen species(ROS),malondialdehyde(MDA),and labile iron pool.Transmission electron microscopy confirmed that the divicine treated HUVECs’mitochondria had shrunk.Importantly,the administration of desferrioxamine,an iron chelator,to the divicine treated HUVECs significantly reduced iron overload and cell death and decreased cellular ROS and MDA.These results demonstrated that divicine could cause damaging of endothelial cells,and ferroptosis might be involved in divicine induced HUVECs injury,reminding long term ingestion of fava beans might be harmful to vascular system,especially for those suffering from glucose 6-phosphate dehydrogenase deficiency.
