Attenuation of cadmium-induced necrotic cell death by necrostatin-1: Potential necrostatin-1 acting sites

• Published in: Toxicology and applied pharmacology 2009-03, Vol.235 (2), p.153-162
• Main Authors: Hsu, Tzu-Sheng, Yang, Pei-Ming, Tsai, Jia-Shiuan, Lin, Lih-Yuan
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; Animals; APOPTOSIS; Apoptosis - drug effects; Biological and medical sciences; CADMIUM; Cadmium Poisoning - pathology; Cadmium Poisoning - prevention & control; CALCIUM; Calcium Signaling - drug effects; Calpain - metabolism; Cell Death - drug effects; Cell Line; Chelating Agents - pharmacology; Chemical and industrial products toxicology. Toxic occupational diseases; CHO Cells; Cricetinae; Cricetulus; DEATH; Electrophoretic Mobility Shift Assay; HAMSTERS; Humans; Imidazoles - pharmacology; Indoles - pharmacology; Medical sciences; Membrane Potentials - drug effects; Metals and various inorganic compounds; MITOCHONDRIA; Mitochondria - drug effects; NECROSIS; Necrostatin-1; NF-kappa B - genetics; NF-kappa B - physiology; NF-κB; OVARIES; Propidium; Reactive Oxygen Species - metabolism; RECEPTORS; Receptors, Drug - drug effects; RIP1; Toxicology; Transfection; U937 Cells
• ISSN: 0041-008X; 1096-0333
• DOI: 10.1016/j.taap.2008.12.012

Cadmium (Cd) induces necrotic death in Chinese hamster ovary (CHO) K1 cells and we have established the responsible signaling pathway. Reportedly, necrostatin-1 (Nec-1) rescues cells from necrotic death by mediating through the death domain receptor (DR) signaling pathway. We show here that Nec-1 also effectively attenuates necrotic death triggered by Cd. Two other treatments that cause necrotic cell death, one can (z-VAD-fmk/TNF-α on U937 cells) and the other cannot (etherynic acid (EA) on DLD-1 cells) be rescued by Nec-1, were also studied in parallel for comparison. Results show that Nec-1 is ineffectual in modulating intracellular calcium contents, calpain activity (a downstream protease), or reactive oxygen species production. It can counteract the reduction in mitochondrial membrane potential (MMP) caused by treating CHO K1 or U937 cells with necrosis-inducing agent. However, this effect was not found in EA-treated DLD-1 cells. Notably, Nec-1 elevates NF-κB activity in the presence or absence of necrosis-inducing agents. Our study shows that, in addition to DR-mediated necrosis, Nec-1 is effective in attenuating Cd-induced necrosis. It rescues cells with reduced MMP implying that mitochondrion is its major acting site.