Rotenone is a specific inhibitor of the NADH dehydrogenase complex. In mitochondria, rotenone inhibitsthe oxidation of NADH to NAD, thereby blocking the oxidation of NAD and the substrates such asglutamate, alpha-ketoglutarate, and pyruvate. Rotenone also inhibits the mitochondrial respiratory chainbetween diphosphopyridine nucleotide and flavine.2, 4-Dinitrophenol – (DNP) is lipophilic weak acids that pick up a proton, transport across the mitochondrialinner membrane into the matrix, deprotonate, then exit as anions before repeating the catalytic cycle,and dissipating the proton gradient. In this situation, electrons continue to pass through the electrontransport system, reduce oxygen to water and metabolic rate, heat are increased, but ATP is lesssynthesized in this process.The macrolide antibiotic - oligomycin binds to the surface of the c8-10 ring of the Fo domain of ATPsynthase, making contact with two neighboring molecules and blocking proton flow, which explains theinhibitory effect on ATP synthesis. Intraperitoneal injection of oligomycin into the rat (0.5 mg per kg)reduces the oxygen consumption by about 50%; decreases ATP production by the aerobic pathway andincreases formation of lactate in blood serum. These changes may cause a decelerated metabolism andan increased formation of free radicals or ROS in membranes.