The Postulated Hepatotoxic Metabolite of Methimazole Causes Mitochondrial Dysfunction and Energy Metabolism Disturbances in Liver

Boldine Supplementation Regulates Mitochondrial Function and Oxidative Stress in a Rat Model of Hepatotoxicity

“…Reactive oxygen species in the liver was assessed using DCF-DA as a ROS probe. 23,24 Briefly, liver tissue samples were homogenized in the ice-cooled Tris-HCl buffer (40 mM, 4ºC, pH = 7.4) (1:10 w/v). Aliquots (100 µl) of the resulted tissue homogenate were mixed with Tris-HCl buffer (1 mL) and DCF-DA (Final concentration of 10 µM).…”

Taurine mitigates cirrhosis-associated heart injury through mitochondrial-dependent and antioxidative mechanisms

“…In the current study, the mitochondrial fractions (1 mg protein/ml) were incubated with rhodamine 123 (final concentration of 10 µM, 15 min, in the dark, with continuous shaking) [ 48 , 75 ]. Afterward, samples were centrifuged (16,000 g, 1 min, 4°C), and the fluorescence intensity of the supernatant was monitored (FLUOstar Omega multifunctional microplate reader, BMG Labtech Inc., Germany, λ excitation = 485 nm and λ emission = 525 nm) [ 62 , 76 ].…”

“…Based on a previously reported protocol, the changes in the light scattering of isolated mitochondria samples at λ = 540 nm was used as an estimate of the mitochondrial swelling and permeabilization [ 47 , 62 , 76 ]. Briefly, isolated heart mitochondria (0.5 mg protein/ml) were suspended in swelling buffer (250 mM sucrose, 2 mM HEPES, 0.5 mM KH 2 PO 4 , pH = 7.2) [ 77 ].…”

“…Then, 100 µl of mitochondrial suspension was transferred to a 96-well plate, and the mitochondrial permeability transition was initiated by adding calcium chloride (final concentration 50 µM). The optical density at λ = 540 nm was monitored for 30 minutes (EPOCH plate reader, Bio-Tek Instruments, Highland Park, USA) [ 76 , 78 ]. A decrease in the absorbance is associated with an increase in mitochondrial volume and organelle swelling [ 62 , 76 ].…”

Propylthiouracil-induced mitochondrial dysfunction in liver and its relevance to drug-induced hepatotoxicity

“…Light absorbance (OD540 nm) was assessed using a FLUOstar Omega ® multifunctional fluorescent microplate reader with constant shaking (100 rpm orbital shaking before each read) and temperature (30ºC). 23,34 A decrease in light absorbance is directly associated with an increase in mitochondrial volume.…”