Peroxynitrite dominates sodium nitroprusside-induced apoptosis in human hepatocellular carcinoma cells
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Ying-Yao Quan1, Yu-Hong Liu1, Chun-Mei Lin1, Xiao-Ping Wang1, Tong-Sheng Chen2
1Department of Pain Management, The First Affiliated Hospital of Jinan University, Guangzhou, China
2MOE Key Laboratory of Laser Life Science and College of Life Science, South China Normal University, Guangzhou, China
Xiao-Ping Wang, email: [email protected]
Keywords: apoptosis, HepG2 cells, nitric oxide, peroxynitrite, sodium nitroprusside
Received: December 20, 2016 Accepted: March 03, 2017 Published: March 13, 2017
This study aims to explore which radicals dominate sodium nitroprusside (SNP)-induced cytotoxicity in human hepatocellular carcinoma (HCC) cells (HepG2 and Hep3B). Exposure of SNP to cell medium produced abundant nitric oxide (NO), superoxide anion (O2•−), hydrogen peroxide (H2O2) and iron ions. SNP potently induced caspases activation, mitochondrial membrane permeabilization and apoptosis in HCC cells. In Hep3B cells, pretreatment with NO scavenger (PTIO) did not prevent SNP-induced cytotoxicity. However, in HepG2 cells, SNP-induced cytotoxicity was prevented significantly by pretreatment with PTIO and O2•− scavenger, and especially was almost completely blocked by pretreatment with FeTPPS (peroxynitrite scavenger). In contrast, although H2O2 scavenger potently scavenged SNP-induced H2O2 production, it did not prevent SNP-induced cytotoxicity in HepG2 cells. In addition, pretreatment with DFO (iron ions chelator) and iron-saturated DFO respectively completely prevented SNP-induced cytotoxicity in HepG2 cells. Collectively, peroxynitrite from the reaction between NO and O2•− elicited from SNP dominates the SNP-induced apoptosis of HepG2 cells, in which both iron ions and H2O2 are not involved.
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