Deciphering the regulatory mechanisms of honeysuckle (Lonicera japonica) on lipopolysaccharide-induced inflammatory responses in loach (Misgurnus anguillicaudatus) through small RNA sequencing

This study investigates the immunomodulatory effects of <i>Lonicera japonica</i> Thunb. extract, a perennial semi-evergreen vine of the Caprifoliaceae family, on lipopolysaccharide (LPS)-induced immune responses in loach (<i>Misgurnus anguillicaudatus</i>) through miRNA regulatory mechanisms. Small RNA libraries constructed from hepatic tissues of LPS-challenged (CK) and Lonicera japonica-treated (LJ) groups yielded 139.6 million clean reads with characteristic 21-23 nucleotide length distribution. Abundance analysis revealed significant differential expression patterns within the let-7 family: miR-let-7-9, miR-let-7-6, and miR-let-7-18 exhibited higher abundance in the infection group, whereas miR-let-7-1, miR-let-7-17, and miR-let-7-16 showed elevated abundance in the treatment group. Comparative profiling identified 55 differentially expressed miRNAs (41 upregulated, 14 downregulated), with animal-undef-351, animal-mir-21-6, and animal-undef-603 demonstrating the most significant expression differences (P<0.01). KEGG enrichment analysis highlighted predominant involvement in sphingolipid signaling pathway, glycerophospholipid metabolism, T cell receptor signaling pathway, and TNF signaling pathway. GO analysis revealed significant enrichment in biological processes related to glycosylation, cellular components of transcription regulator complexes, and molecular functions associated with double-stranded DNA binding. These findings demonstrate that <i>L. japonica</i> alleviates LPS-induced inflammation by modulating miRNA expression networks, particularly through sphingolipid metabolism and TNF signaling pathways, providing novel molecular evolutionary insights into the immunoregulatory mechanisms of traditional Chinese medicine.