2.6. Quantitative Real-Time PCR (qRT-PCR) Analysis of the Kidney

The expression levels of TNF-α, IL-1β, IL-6, and NF-κB P65 were analyzed via qRT-PCR. Briefly, total RNA was isolated from renal tissues with TRIzol reagent (Thermo Fisher Scientific, USA) according to the manufacturer's protocol and reversely transcribed into cDNA using reverse transcription kits (Takara Bio, Japan). Real-time PCR analysis was performed with a QuantiTect™ SYBR Green PCR (Takara Bio, Japan) according to the manufacturer's instructions. The primers (Table 1) were synthesized by Shanghai Biological Engineering (Shanghai, China). Relative quantification of the target gene expression levels was conducted using the 2^−∆∆Ct method.

Sequences of the primers used for PCR [15, 16].