Quantitative real-time reverse transcriptase PCR

Small pieces of atrial and ventricular tissue were placed into liquid nitrogen for snap freezing and later stored in − 80 °C freezer until analysis. Four rats in each group were chosen for heart mRNA expression analysis. Total RNA was prepared using TRI Reagent (Sigma-Adrich, St Louis, MO), then reverse transcribed (Invitrogen, Carlsbad, CA). Quantitative real-time RT-PCR was performed using an ABI 7500 real-time system (Applied Biosystems, Foster City, CA) and TaqMan Universal Master Mix II with TaqMan probes (Applied Biosystems, Foster City, CA): PPARα, Rn00566193_m1; ACADM, Rn00566390_m1; TNF-α, Rn01525859_g1; ANP, Rn00664637_g1 and BNP, Rn00580641_m1.