Differential mechanistic investigation of protective effects from imperatorin and sec-O-glucosylhamaudol against arsenic trioxide-induced cytotoxicity in vitro

• Published in: Toxicology in vitro 2016-12, Vol.37, p.97-105
• Main Authors: Hu, Liufang, Sun, Jianhui, Li, Hongmei, Wang, Lifang, Wei, Yuna, Wang, Ying, Zhu, Yaying, Huo, Hairu, Tan, Yuqing
• Format: Article
• Language: English
• Subjects: Acridine orange; Acute promyeloid leukemia; Animals; Antioxidants; Apoptosis; Apoptosis - drug effects; Arsenic; Arsenic Trioxide; Arsenicals; As2O3-induced cytotoxicity; Calcium; Calcium (intracellular); Calcium - metabolism; Cardiotonic Agents - pharmacology; Cardiotoxicity; Caspase; Caspase 3 - metabolism; Caspase-3; Cell death; Cell Line; Cell Survival - drug effects; Chromones - pharmacology; Cytoprotection; Cytotoxicity; Deoxyribonucleic acid; DNA; DNA fragmentation; Enzymes; Ethidium bromide; Experimental methods; Furocoumarins - pharmacology; Gene expression; Heme Oxygenase (Decyclizing) - genetics; Imperatorin; Ion channels; L-Lactate dehydrogenase; L-Lactate Dehydrogenase - metabolism; Lactate dehydrogenase; Lactic acid; Leukemia; Mechanisms; NAD(P)H Dehydrogenase (Quinone) - genetics; NF-E2-Related Factor 2 - genetics; Oxides - toxicity; Promyeloid leukemia; Proteins; Rats; Reactive oxygen species; Reactive Oxygen Species - metabolism; RNA, Messenger - metabolism; sec-O-glucosylhamaudol; Toxicity; Western blotting
• ISSN: 0887-2333; 1879-3177
• DOI: 10.1016/j.tiv.2016.09.002

The clinical use of arsenic trioxide (As2O3) for treating acute promyelocytic leukemia (APL) is limited due to its severe cardiotoxicity. The possible mechanisms of As2O3-induced cardiotoxicity include DNA fragmentation, reactive oxygen species (ROS) generation, cardiac ion channel changes and apoptosis. The present study is designed to investigate the protective effects of imperatorin and sec-O-glucosylhamaudol and to explore their mechanistic involvement in As2O3-induced cytotoxicity. Cell viability assay, Lactate dehydrogenase (LDH) release, Acridine orange/ethidium bromide (AO/EB) double staining, Caspase-3 activity assay, ROS generation, cellular calcium levels, mRNA expression levels by qRT-PCR and protein expression levels by Western blotting were measured in H9c2 cells in combination with As2O3 and imperatorin or sec-O-glucosylhamaudol. We observed that H9c2 cells treated with imperatorin or sec-O-glucosylhamaudol were more resistant to As2O3-induced cell death. Both imperatorin and sec-O-glucosylhamaudol reduced H9c2 cell apoptosis, but both imperatorin and sec-O-glucosylhamaudol had no effects on Caspase-3 activity and intracellular calcium accumulation. Furthermore, imperatorin was capable of suppressing ROS generation, while sec-O-glucosylhamaudol did not show this effect. Moreover, imperatorin and sec-O-glucosylhamaudol triggered Nrf2 activation, which resulted in upregulation of downstream phase II metabolic enzymes and antioxidant protein/enzyme, probably offering cellular protection to As2O3-induced cardiotoxicity via the Nrf2 signal pathway. Imperatorin and sec-O-glucosylhamaudol can ameliorate As2O3-induced cytotoxicity and apoptosis in H9c2 cells, the mechanisms probably related to antioxidation. As2O3 in combination with imperatorin or sec-O-glucosylhamaudol could be considered as a novel strategy to expand the clinical application of As2O3. •The new pharmacological action of traditional Chinese medicine Radix Saposhnikoviae•Imperatorin can ameliorate As2O3-induced cytotoxicity and apoptosis in H9c2 cells.•sec-O-glucosylhamaudol can ameliorate As2O3-induced cytotoxicity and apoptosis in H9c2 cells.•A novel strategy to expand the clinical application of As2O3