Study of the Stability and Anti-Inflammatory Activity of Paeonol–Oleanolic Acid Liposomes by Microfluidic Technology

(1) Background: This study used a microfluidic approach to prepare paeonol (PAE) liposomes with oleanolic acid (PAE-ONLs) instead of cholesterol (PAE-CNLs), aiming to reduce cholesterol levels and enhance stability and anti-inflammatory activity. (2) Methods: The liposome formula was optimized, characterized, and tested for anti-inflammatory activities in zebrafish and RAW 264.7 macrophages, utilizing various stability and molecular interaction methods. (3) Results: The best PAE-ONL preparation conditions were 10.25 mg/mL of soy lecithin, 0.82 mg/mL of oleanolic acid, and 0.22% (wt%) of Tween 80, with an <i>EE</i> of 64.61 ± 0.42%. TEM confirmed the uniform spherical morphology, and FTIR confirmed that oleanolic acid was incorporated into the liposomes. PAE-ONLs showed better stabilities than PAE-CNLs. Molecular interaction results revealed that PAE-ONLs achieved a greater energy reduction, reaching -85.07 kJ/mol vs. the -62.64 kJ/mol of PAE-CNLs, with stable hydrogen bonding interactions. PAE-ONLs significantly reduced inflammatory cell migration in zebrafish and decreased NO, TNF-α, IL-6, and IL-1β levels in LPS-induced RAW 264.7 macrophages at 20 μg/mL. A network pharmacology analysis showed that oleanolic acid and paeonol interacted with 45 and 11 anti-inflammatory targets, respectively, and their combination in PAE-ONLs enhanced their anti-inflammatory coverage. (4) Conclusions: PAE-ONLs, utilizing oleanolic acid as a cholesterol substitute, exhibit enhanced stability and superior anti-inflammatory effects.