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. Author manuscript; available in PMC: 2017 Nov 15.
Published in final edited form as: Int J Cardiol. 2016 Aug 13;223:923–935. doi: 10.1016/j.ijcard.2016.08.233

Figure 11. Diclofenac induces mitochondrial dysfunction and mitochondrial complex I inhibitor induces ROS generation and cell death and decreases proteasome activity of H9c2 cells.

Figure 11

A, Effect of DIC on complex I activity in mitochondria isolated from mouse hearts. Rotenone (ROT), a known inhibitor of complex I, was used as a control. B, Effect of DIC on complex II activity in mitochondria isolated from mouse hearts. Thenoyltrifluoroacetone (TTFA), a complex II inhibitor, was used as a control. C, Effect of DIC on complex III activity in mitochondria isolated from mouse hearts. The complex III inhibitor, Antimycin A, was used as a control. D, Effect of DIC on mitochondrial membrane potential measured in H9c2 cells. Rotenone (ROT) and 2,4-dinitrophenylhydrazine (DNPH) were used as controls. E, ROS levels were determined in the presence of DIC and DIC + Mito-Tempol, a mitochondrial ROS scavenger. F, ROS generation, cell viability and proteasome β5 activity of H9c2 cells in the presence of rotenone. H9c2 cells were plated onto 96 plates at 4000 cells/well and H2DCFDA and Alamar blue was utilized to determine ROS generation and cell viability as described in the Methods. ROS generation in H9c2 cells in the presence of rotenone was determined both by fluorimetry and fluorescent microscopy studies. The fluorescent images are a representative of 3 or more independent experiments. Effect of rotenone on proteasome β5 activity was also measured. ROT-rotenone. Results are mean ± SD (n=4–6)* P < 0.05, ** P < 0.001.