*4.13. Quantitative Reverse-Transcription PCR (qRT-PCR) Analyses*

The total RNA from each sample was isolated using the Trizol reagent (Sango Shanghai, China), and the first strand cDNA was synthesized using the StarScript II First-strand cDNA Synthesis Mix With gDNA Remover (Genstar) according to the manufacturer's instructions. Quantitative reverse-transcription PCR (qRT-PCR) was conducted to determine the mRNA levels of the IL-1β, IL-6, TNF-α, Bax, Caspase-3, and Bcl-2 (the primer sequences are shown in Table 4), the GAPDH gene was used as an internal control [8,30,97]. Real-time PCR reactions were performed on a CFX Real-time system (CFX96, Bio-Rad, Hercules, CA, USA). All of the samples were analyzed in triplicate, and the 2−ΔΔCt method was used to analyze gene expression levels.

**Table 4.** Primer sequences for qRT-PCR analyses.

