RNA sequencing reveals the emerging role of bronchoalveolar lavage fluid exosome lncRNAs in acute lung injury

Abstract

Background Bronchoalveolar lavage fluid (BALF) exosomes possess different properties in different diseases, which are mediated through microRNAs (miRNAs) and long noncoding RNAs (lncRNAs), among others. By sequencing the differentially expressed lncRNAs in BALF exosomes, we seek potential targets for the diagnosis and treatment of acute lung injury (ALI). Methods Considering that human and rat genes are about 80% similar, ALI was induced using lipopolysaccharide in six male Wistar rats, with six rats as control (all weighing 200 ± 20 g and aged 6–8 weeks). BALF exosomes were obtained 24 h after ALI. The exosomes in BALF were extracted by ultracentrifugation. The differential expression of BALF exosomal lncRNAs in BALF was analyzed by RNA sequencing. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to predict the functions of differentially expressed lncRNAs, which were confirmed by reverse transcription–polymerase chain reaction. Results Compared with the control group, the ALI group displayed a higher wet/dry ratio, tumor necrosis factor-α levels, and interleukin-6 levels (all P < 0.001). The airway injection of exosomes in rats led to significant infiltration by neutrophils. A total of 2,958 differentially expressed exosomal lncRNAs were identified, including 2,524 upregulated and 434 downregulated ones. Five lncRNAs confirmed the reliability of the sequencing data. The top three GO functions were phagocytic vesicle membrane, regulation of receptor biosynthesis process, and I-SMAD binding. Salmonella infection, Toll-like receptor signaling pathway, and osteoclast differentiation were the most enriched KEGG pathways. The lncRNA–miRNA interaction network of the five confirmed lncRNAs could be predicted using miRDB. Conclusions BALF-derived exosomes play an important role in ALI development and help identify potential therapeutic targets related to ALI.

RANK signaling in osteoclast precursors results in a more permissive epigenetic landscape and sexually divergent patterns of gene expression

“…By contrast, expression of TLR1, TLR2, and TLR9 was higher in male macrophages. These findings suggest that male naïve macrophages are more sensitive to TLR ligands and agree with prior findings of sexually dimorphic responses to inflammatory signals and induction of TLR expression in conjunction with pro-osteoclastogenic pathways ( Marriott, Bost & Huet-Hudson, 2006 ; Li et al, 2009 ; Barcena et al, 2021 ; Chen, Lainez & Coss, 2021 ; Song et al, 2022 ). The potency of some TLR ligands, such as bacterial lipopolysaccharide (LPS), to stimulate osteoclastogenesis of osteoclast precursors pre-committed with RANKL is well established ( Liu et al, 2009 ).…”

Network Pharmacology Analysis of the Therapeutic Potential of Colchicine in Acute Lung Injury

“…The mice were randomized into three groups: ALI group, colchicine treatment group (ALI with colchicine treatment), and control group. Group sizes were determined based on previous research [ 34 , 35 ]. To establish the ALI model, mice were exposed to 0.8 mg/ml nebulized lipopolysaccharide ( Escherichia coli O111: B4, Sigma, USA) diluted in 0.9% NaCl for 30 minutes, consistent with previous studies [ 36 , 37 ].…”