Background: MicroRNA-144 (miR-144) has been implicated in inflammation and glucocorticoid (GC) receptor regulation, but its role in GC sensitivity during acute respiratory distress syndrome (ARDS) is unclear. This study aimed to investigate the impact of miR-144 on GC therapeutic efficacy, elucidate its underlying molecular mechanisms in ARDS, and validate its clinical application value. Methods: First, we established a sepsis-induced ARDS rat model with five groups (n = 6 each): control, model, GC, GC + miR-144 mimic, or GC + miR-144 inhibitor. Body weight, inflammatory markers, lung wet/dry ratio, histopathology, and miR-144/GR alpha/GR(3/NF-kappa B mRNA were assessed after the 5-day treatment. Second, we prospectively enrolled 63 ARDS patients receiving methylprednisolone and classified them as GC-sensitive (n = 43) or GC-resistant (n = 20) based on clinical outcomes. The correlations of baseline miR-144, GR alpha, GR(3, and NF kappa B mRNA levels in patient blood were analyzed, and the predictive value of miR-144 for GC sensitivity was evaluated. Results: In rats, miR-144 inhibitor enhanced GC efficacy (improved body weight recovery, reduced lung edema and inflammation), while mimic attenuated these effects. Mechanistically, in rat lung tissue, sepsis stimulation increased miR-144, GR(3, and NF-kappa B while reducing GR alpha expression; GC reversed these changes; miR-144 inhibitor further reduced GR(3/NF-kappa B, whereas mimic antagonized this without affecting GR alpha. In patients, GC-resistant individuals showed significantly higher miR-144, GR(3, and NF-kappa B levels than sensitive ones. Strong positive correlations were observed between miR-144 and both GR(3 and NF-kappa B, and miR-144 effectively predicted GC resistance in ARDS. Conclusions: MiR-144 regulates GC sensitivity via the GR(3/NF-kappa B axis in ARDS. Baseline miR-144 level is a promising biomarker for predicting GC response, providing a basis for individualized treatment.
Seven 11α-hydroxylated lithocholic acid (LCA) derivatives were produced via catalytic biotransformation by Aspergillus ochraceus CGMCC 3.5324, including four previously unreported compounds. All isolates were structurally elucidated using NMR spectroscopy and HR-ESI-MS analyses. A. ochraceus efficiently catalyzed multiple structural modifications on the LCA skeleton, including hydroxylation, methyl esterification, and acetylation, thereby expanding the structural diversity of LCA derivatives. Furthermore, the anti-aging activities of these biotransformed metabolites were evaluated in a Caenorhabditis elegans model by measuring lifespan and locomotor behavior. Most transformed derivatives exhibited superior anti-aging potency relative to the parent LCA. Methyl and acetyl modifications markedly improved nematode lifespan and locomotor performance, while C-12β hydroxylation reduced the bioactivity of LCA derivatives. These findings demonstrate that microbial biotransformation is a robust strategy for the structural diversification and bioactivity optimization of LCA.
Soluble epoxide hydrolase (sEH) is a promising therapeutic target for inflammation-related disorders; however, current synthetic inhibitors are limited due to pharmacokinetic constraints. Marine steroidal metabolites offer a potential source of natural sEH inhibitors. Therefore, this study aims to investigate the sEH inhibitory activity of compounds isolated from the starfish Patiria pectinifera. Six steroidal compounds (1-6) were isolated and evaluated using enzymatic assays, kinetic analysis, molecular docking, and molecular dynamics simulations. Compounds 1 and 2 demonstrated potent sEH inhibition (IC50 = 16.10 +/- 2.43 mu M and 4.13 +/- 1.24 mu M, respectively), while compounds 3-6 were inactive. Kinetic studies showed mixed-type inhibition, with lower Kis than Ki values, indicating preferential binding to the enzyme-substrate complex. Computational analyses revealed that compounds 1 and 2 bind to distinct sites within the C-terminal domain with higher affinity than that of the native substrate, inducing conformational changes consistent with allosteric modulation. These findings indicate compounds 1 and 2 as promising allosteric sEH inhibitors, highlighting the potential of P. pectinifera steroids as anti-inflammatory agents.
