2.3. Quantitative Real-Time Polymerase Chain Reaction

Total RNA from liver was extracted using RNeasy Mini Kit (Milan, Italy) according to the manufacturer’s instructions and quantified using a NanoDrop Spectrophotometer at 260 nm. Total RNA (up to 1 ug) was reverse transcribed using iScript cDNA synthesis kit (Bio-Rad, Hercules, CA, USA). The expression of transforming growth factor-beta1 (TGFβ-1), collagen type 1a1 (Col1a1), superoxide dismutase 1 (SOD-1), superoxide dismutase 2 (SOD-2), glutathione peroxidase 1 (GPx-1), catalase, tumor necrosis factor α (TNFα), interleukin-6 (IL-6), interleukin-1β (IL-1β), interleukin 10 (IL-10), Matrix Metallopeptidase 2 (MMP-2), Matrix Metallopeptidase 9 (MMP-9), tissue inhibitor of Metalloproteinase-1 (TIMP-1), and Serpinb3 was determined using SYBR Green Mastermix (Bio-Rad, Hercules, CA, USA) and performed in a CFX Real-Time PCR Detection Systems (Bio-Rad, Hercules, CA, USA) using the primer sequences shown in Table 1. Actin was used as a housekeeping gene. The relative expression was generated for each sample by calculating 2^−ΔΔCt.

List of primers used in mice liver tissues.