Using Janus Green B to Study Paraquat Toxicity in Rat Liver Mitochondria

作者：Mahmoud Ghazi‐Khansari, Afshin Mohammadi‐Bardbori, Mir‐Jamal Hosseini · 发表于：Annals of the New York Academy of Sciences · 年份：2006 · DOI：10.1196/annals.1378.010 · 被引用次数：78 · 研究领域：Paraquat toxicity studies and treatments、Protein Interaction Studies and Fluorescence Analysis、Heme Oxygenase-1 and Carbon Monoxide

Janus green B (JG-B) dye is used for vital staining of mitochondria and its reduction and oxidation shows the electron transfer chain alteration. The defect in electron transfer chain of mitochondria by paraquat is linked to free radical formation. In this present study we compared the abilities of different angiotensin-converting enzyme inhibitors, captopril (a thiol ACEi), enalapril, and lisinopril (two nonthiol ACEi) on mitochondria toxicity due to paraquat. The rat liver mitochondria were first isolated by centrifuge (at 4 degrees C at a speed of 7,000 g) in a mixture of 0.25 M saccharose solution and 0.05 M Tris buffer. Various concentrations of paraquat (1, 5, 10 mM), enalapril (0.25, 0.5, 1 mM), lisinopril (0.01, 0.05, 0.1 mM), and captopril (0.08, 0.1, 1 mM) on the mitochondria isolated from the liver with respect to time were investigated. Paraquat at a concentration of 5 mM was determined to be significantly different compared to control values (P<0.05) and captopril at a concentration of 0.08 mM, lisinopril (0.01 mM), and enalapril (0.25 mM) were found not to be significantly different from controls as found by spectroscopy at wavelength of 607 nm. Simultaneous treatment of mitochondria with captopril (0.08 mM) and paraquat (5 mM) significantly ameliorates the mitochondria toxicity of paraquat (5 mM) alone (P<0.05). Our results show that captopril is a more effective antioxidant than the nonthiol ACEi. Lisinopril (0.01 mM) and enalapril (0.25 mM) did not significan...