Characterization of andrographolide-loaded exosome-like nanoparticles from Nauclea officinalis and their modulation of inflammatory and lipid metabolic pathologies in WRL68 cells.

Andrographolide (AG), a natural diterpenoid compound derived from Andrographis paniculata, exhibits potential against non-alcoholic steatohepatitis (NASH). However, its therapeutic utility is limited by poor solubility, short half-life, and low bioavailability. This study aimed to enhance AG's efficacy by encapsulating it into exosome-like nanoparticles (ELNs) isolated from Nauclea officinalis (N. officinalis, Rubiaceae family) and evaluating its anti-NASH activity in WRL68 cells. This study isolated Nauclea officinalis-derived ELNs (N-ELNs) via ultracentrifugation. Loganin, andrographolide, and asiatic acid were detected in N-ELNs using the HPLC method. RNA sequencing and target gene analysis identified miRNAs in N-ELNs and their cross-kingdom targets in the human genome. AG-loaded N-ELNs (AG-N-ELNs) were prepared using a passive loading method and characterized by transmission electron microscopy, nanoparticle tracking analyzer, and high-performance liquid chromatography. Their effects were tested in free fatty acid (FFA)-exposed WRL68 cells. As for the results, AG suppressed Nuclear Factor kappa-B p50 (NF-κB p50) activation, downregulated NLRP3 expression, and reduced pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) in WRL68 cells. It also restored mitochondrial membrane potential and decreased reactive oxygen species (ROS). N-ELNs targeted the ACC/ CPT1 axis to alleviate lipid deposition, reducing total cholesterol and triglyceride levels. AG-N-ELNs outperformed both AG and N-ELNs individually, demonstrating superior anti-NASH activity through synergistic effects of AG's anti-inflammatory/antioxidant properties and N-ELNs' miRNA-mediated metabolic regulation. In conclusion, AG-N-ELNs effectively alleviate NASH-like pathological features in vitro through multi-target regulation, offering a promising strategy for NASH treatment. Further in vivo validation and formulation optimization are warranted.