Transcriptomic Analysis of the Adrenal Glands, Heart, and Pancreas in Rats Following Benzalkonium Chloride Inhalation

Transcriptomic profiling was conducted on the adrenal glands, heart, and pancreas 24 hours after the 10th intratracheal instillation of benzalkonium chloride in rats. Total RNA was extracted using TRIzol and purified with the RNeasy Mini kit. High-quality RNA (RIN > 9) was used to construct sequencing libraries with the TruSeq Stranded mRNA kit, followed by sequencing on a NovaSeq 6000 platform. Overall design: Quantitative RNA sequencing was performed with adrenal glands, heart, and pancreas at 24 h after the 10th intratracheal instillation for benzalkonium bhloride (BKC). Collected organs were added 1 mL of TRIzol reagent (Invitrogen Life Technologies, Carlsbad, CA, USA) per 50 mg of organ tissues. The paired adrenal glands per animal were pooled, and the heart was dissected in the left ventricular area. Samples weighed approximately 50 mg and were stored in TRIzol at -80? in a deep freezer until RNA purification. Total RNA was isolated using a RNeasy Mini kit (Qiagen, Valencia, CA, USA), and the RNA integrity was analyzed using a Bioanalyzer (Agilent, Santa Clara, CA, USA). RNA samples with RNA integrity values greater than 9 were selected to prepare the mRNA sequencing library. The library was prepared using a TruSeq Stranded mRNA library kit (Illumina, San Diego, CA, USA), and sequencing was performed using a NovaSeq 6000 (Illumina).