2.8. Determination of Hepatocyte Apoptosis by the TUNEL Method

The apoptotic cells were assessed using a commercial assay kit (Roche, IN, USA) [6]. The liver tissue sections underwent strict dewaxing. Next, 50 μL of TUNEL reaction mixture was added, and the sections were incubated at 37°C for 60 min in a dark humidified atmosphere. After stained again, apoptotic hepatocytes were observed under the fluorescence microscope at high-magnification (400x) field of vision. Five fields of vision were randomly selected to observe and calculate the apoptosis index and hence evaluate the hepatocyte apoptosis.