Transcriptomic study of Xiao-Qing-Long-Tang in the treatment of OVA-induced allergic rhinitis in mice
Nasal mucosa samples from mice in the Ctrl, AR model, and XQLT groups (n = 3) were sent to Majorbio Biotechnology Co., Ltd. (Shanghai, China), where total RNA from Nasal mucosa tissues was extracted using Trizol (Invitrogen, Carlsbad, CA, USA). Agilent 2100 and Nanodrop were used to determine RNA quality. After enriching the mRNA using magnetic beads and Oligo(dT), short fragments of the mRNA were created by adding fragmentation buffer. Random hexamers were used for reverse transcription, and buffer, dNTPs, and DNA polymerase I were added to create two-stranded cDNA. After a series of purifications, repair, and amplification, the cDNA library was constructed. Using the Illumina high-throughput sequencing platform (HiSeqTM2500), the prepared libraries were sequenced using the PE150 strategy and then compared to the reference genome sequence using STAR (v2.5.2b). HTSeq software was used to analyze the gene expression levels of each sample to obtain FPKM (Fragments per kilobase of exon model per million mapped reads) values. Next, DESeq (1.10.1) was used to determine the differentially expressed genes based on |log2(FoldChange)| > 1 and padj<0.05. GOSeq (v1.22) and KOBAS (v2.0) were used to perform differential gene GO enrichment and KEGG enrichment analysis on the corrected p-value<0.05.