ISSN:
0378-8741
Language:
English
Titel der Quelle:
Journal of ethnopharmacology : an interdisciplinary journal devoted to bioscientific research on indigenous drugs
Publ. der Quelle:
Shannon : Elsevier Science Ireland
Angaben zur Quelle:
Vol. 174 (2015), p. 331
DDC:
610
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. Additionally, cell cycle analysis was performed to detect the effects of 3EZ,20Ac-ingenol on cell cycle in IEC-6 cells by flow cytometry. The study showed that 3EZ,20Ac-ingenol significantly reduced IEC-6 cells viability in a dose-dependent manner and the IC50 value was 5.74μg/mL. Consistently, 3EZ,20Ac-ingenol could elevate reactive oxygen species (ROS), disrupt mitochondrial membrane potential (MMP), induce the release of cytochrome c from mitochondria to cytosol, enhance the expressions of Bax, AIF and Apaf-1, suppress the expression of Bcl-2, then activate caspase-3, 8, 9 cascade, and subsequently result in apoptosis. Additionally, 3EZ,20Ac-ingenol also could cause G2/M phase arrest in IEC-6 cells. The results indicated that 3EZ,20Ac-ingenol induced the cytotoxicity of IEC-6 cells depends on induction of cell apoptosis via mitochondrial pathway and cell cycle arrest.
Note:
Copyright: Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.
DOI:
10.1016/j.jep.2015.08.036
URL:
http://www.ncbi.nlm.nih.gov/pubmed/26318745
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