Role of type-II pathway in apoptotic cell death induction by photosensitized CDRI-97/78 under ambient exposure of UV-B
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- 作者：Ashish Dwivedi ; Manish Kumar Pal ; Amit Kumar Tripathi ; Neera Yadav ; Syed Faiz Mujtaba ; M.C. Pant ; Shio Kumar Singh ; Durga Prasad Mishra ; Ratan Singh Ray ; B.H. Manjunatha Prabhu
- 关键词：Antimalarial ; LC&ndash ; MS/MS ; UV-B ; DNA damage ; Mitochondrial membrane destabilization
- 刊名：Toxicology Letters
- 出版年：2013
- 出版时间：24 October, 2013
- 年：2013
- 卷：222
- 期：2
- 页码：122-131
- 全文大小：3408 K
文摘
Novel trioxane 97/78, developed by Central Drug Research Institute (CDRI), Lucknow has shown promising antimalarial activity. Clinical experience of anti-malarial drugs registered the occurrence of phototoxicity in patients exposed with sunlight subsequent to medication. Photodegradation study has identified one photo-product up to 4 h under UV-B/Sunlight by LC-MS/MS. UV-B irradiated 97/78 compound produced 1O2 via type-II dependent reaction mechanism, corroborated by its specific quencher. 2¡ä-dGuO degradation and % tail development in photochemical as well as comet test, advocated the genotoxic potential of 97/78. The photocytotoxicity assays (MTT and NRU) on HaCaT cell line revealed the considerable decline in cell viability by 97/78. Cell cycle and Annexin V/PI double stain along with AO/EB demonstrated the G2/M phase arrest and apoptosis. Significant caspase-3 activity was measured in photoexcited 97/78 by colorimetric assay. Fluorescence stain with AO/JC-1 confirmed the lysosomal disruption and mitochondrial membrane destabilization by UV-B irradiated 97/78. Gene expression by RT-PCR showed significant upregulation of p21 and pro-apoptotic Bax, but no change observed in Bcl-2. In conclusion, the study highlights ROS mediated DNA damage, lysosomal and mitochondrial destabilization via upregulation of Bax and activation of caspase-3 which further leads to apoptosis.