Mechanism of NAT10 inhibitor in treating thalamic hemorrhage by integrated serum metabolomics and thalamic transcriptomics

Objective To investigate the mechanism of N-acetyltransferase 10 (NAT10) inhibitor Remodelin in treating thalamic hemorrhage and identify key molecules and metabolites.

Methods Healthy male C57BL/6J mice aged 6 to 8 weeks with specific pathogen-free status were selected as the study subjects. They were randomly divided into control group (C Group, n=9), thalamic hemorrhage group (TH Group, n=9) and Remodelin intervention group (TH+R Group, n=9) using a random number table method. In the TH Group, a thalamic hemorrhage model was established by injecting 0.01 U of type IV collagenase into the right thalamus using a stereotaxic instrument, while C Group received an equivalent volume of saline injection. In the TH+R Group, Remodelin at a dose of 5 mg/kg was administered via intraperitoneal injection 24 h after collagenase injection. At 3 days post-modeling, blood was collected from the eyeballs and allowed to stand for 30 min, after which the supernatant was aspirated for metabolomic detection and analysis. Subsequently, the mice were euthanized, and the right thalamic tissue was harvested for RNA extraction, followed by transcriptome sequencing and RT-qPCR to detect the expression of key genes. The results of transcriptomic and metabolomic analyses were analyzed to screen for differentially expressed genes and metabolites in thalamic hemorrhage under Remodelin intervention. Kyoto Encyclopedia of Genes and Genomes (KEGG) and metabolite-gene network analyses were performed to identify key targets. The binding ability of Remodelin to key genes was validated through molecular docking.

Results A total of 42 common differentially expressed metabolites (CDEMs) and 499 common differentially expressed genes (CDEGs) were identified. KEGG analysis revealed that the cyclic adenosine monophosphate (cAMP) signaling pathway was the commonly enriched pathway for both CDEMs and CDEGs. Network analysis of genes and metabolites within this pathway identified two key genes ATPase sarcoplasmic/endoplasmic reticulum Ca²⁺ transporting 1 (Atp2a1) and proopiomelanocortin (Pomc) and one key metabolite lysophosphatidic acid (LPA). Molecular dockingconfirmed significant binding between Remodelin and the key genes. Compared with the C Group, the expression levels of Atp2a1, Pomc and LPA were decreased in the TH Group, with statistically significant differences (P < 0.05). Compared with the TH Group, the expression levels of Atp2a1, Pomc and LPA were increased in the TH+R Group, with statistically significant differences (P < 0.05).

Conclusion Atp2a1 and Pomc are involved in the therapeutic process of Remodelin in mice with thalamic hemorrhage, and the mechanism may be related to the LPA-associated cAMP pathway.