2.4. Quantification of Genes Expression in Liver Tissue

Total RNA was extracted and purified in the liver tissue using TRIzol reagent (TAKARA, Beijing, China. RNAiso Plus, Code No. 9108). The purity, concentration, and quality of RNA were measured. High-quality RNA was converted to cDNA using PrimeScript™ RT reagent Kit with gDNA Eraser (TAKARA, Beijing China. Code No. RR047A). The SYBR fluorescent dye method (TAKARA, Beijing China. Code No. RR420A) and Agilent Stratagene Mx3000P Real-Time PCR System (Santa Clara, CA, USA) were used to detect the gene expression. The primers used are shown in Supplementary Table S2. The data were calculated using the 2^−ΔΔCt relative quantification method and normalized to β-actin.