Optimization of Hesperidin Nanoparticles as a Promising CYP2E1 and CYP3A11 Regulator in Paracetamol-Intoxicated Mice.

Background: Hesperidin is a flavonoid found in citrus fruits, particularly in the peel and pulp of oranges and lemons. By encapsulating drugs or bioactive compounds within NPs, it's possible to enhance their stability, solubility, and bioavailability. The current investigation aims to optimize hesperidin nanoparticles (Hes-NPs) and evaluate their hepatoprotective and antioxidant effects in paracetamol-intoxicated mice.

Methods: The characteristics of Hes-NPs were elucidated, including morphology, particle size, zeta potential, UV-vis, entrapment efficiency, and FT-IR spectra. Hes-NPs were also tested for their hepatoprotective and antioxidant effects in paracetamol-treated mice. Safety and toxicity assessments are crucial before pharmacological studies. In addition, liver enzymes, oxidative stress, inflammatory biomarkers, and gene expression of CYP2E1 and CYP3A11 were measured. Furthermore, the study examined the molecular docking of hesperidin with the CYP2E1 and CYP3A11 proteins.

Results: The synthesized Hes-NPs were uniform, spherically shaped, and well dispersed, with no aggregation noted, with a size range of 155.12 ± 14.13 nm. The measured zeta potential value of Hes-NPs was -21.57 ± 0.8 mV with a polydispersity index (PDI) of 0.49. Also, the UV spectra of hesperidin are at 220 and 279 nm, and Hes-NPs have strong absorption at 225 and 280 nm. Also, the LD50 of Hes-NPs was 1137.5 mg/kg b.w. Moreover, administering paracetamol-intoxicated mice with Hes-NPs resulted in improved plasma lipid profile (TC, TG, and HDL-C) and liver enzymes (ALT, AST, ALP, and LDH) as well as oxidative stress (GSH, SOD, CAT, Pr-SHs, and MDA) and inflammatory (TNF-α) biomarker levels, as well as attenuated CYP2E1, and CYP3A11 gene expression. In-silicon results proved that hesperidin showed a stronger estimated binding affinity with a ∆G of -8.6 and -10.5 kcal/mol. towards CYP2E1, and CYP3A11 activity, respectively. Our results showed that hesperidin forms hydrogen bonds with amino acid residues through the 9 and 12 bonds of CYP2E1 and CYP3A11, respectively.

Conclusion: Hes-NPs could offer several advantages. It can be designed to specifically target liver cells, minimizing off-target effects, enhancing bioavailability, and shielding hesperidin from degradation in the body. The current results suggest that Hes-NPs may enhance antioxidant defenses and protect against oxidative stress in paracetamol-intoxicated mice.