Abstract
Euphorbia kansui is a traditional Chinese medicine widely used for the treatment of edema, ascite and asthma in China for centuries. However, its serious gastrointestinal toxicity restricted its safe clinical application. 3-O-(2'E,4'Z-decadienoyl)-20-O-acetylingenol (3EZ,20Ac-ingenol), a diterpenoid compound derived from kansui, has obvious gastrointestinal cytotoxicity in cells. Until now, its gastrointestinal cytotoxic mechanism is mostly unknown. This study focused on elucidating the cytotoxic mechanism of 3EZ,20Ac-ingenol in intestinal epithelial cells of rats (IEC-6 cells) to guide safer application of this herb in clinic. 3EZ,20Ac-ingenol was isolated from the EtOAc extract of kansui. Cell morphology was detected by inverted phase contrast microscope and transmission electron microscope (TEM). Cell apoptosis was examined by Annexin V-FITC/PI dual-staining or Hoechst staining. ROS generation was detected with DCFH-DA staining by laser scanning confocal microscope. MMP change was examined with JC-1 staining by high content screening (HCS). Further, the release of cytochrome c, the expressions of Bax, Bcl-2, AIF and Apaf-1 were analyzed by western blot and the activities of caspase-3, 8, 9 were determined by ELISA. Additiona...Continue Reading
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