Tier 3 — preclinical

Fisetin-loaded nanoparticles as a novel approach for cholesterol regulation in hypercholesterolemia: targeting the ASGR1-mediated mTORC1/AMPK pathway

Zhang Z, Chen X, Hu X, Liang J, Huang H, Lu W, Zhu M, Fang M, Yin L, Li W, Zhang S
Journal of Nanobiotechnology 2026 Volume 24, article 312

Bibliography

PubMed
PMID 41761201
PubMed Central
PMC13050004
DOI
10.1186/s12951-026-04181-z
Funding
National Natural Science Foundation of China (grant no. 82473623); Key Scientific Research Project of Colleges and Universities in Henan Province (grant no. 24A330006); Henan Provincial Science and Technology Research Project (grant no. 232102310515); The 2025 Innovative Training Program for College Students of Zhengzhou University (grant no. S202510459171).
Competing interests
The authors declare no competing interests.

Study snapshot

DesignControlled animal-feeding study (high-fat, high-cholesterol diet-induced hypercholesterolemia mouse model) with mechanistic in vitro validation in AML12 hepatocytes, molecular docking and cellular thermal shift assay (CETSA) target-engagement confirmation, plus development and testing of a carboxymethyl chitosan (CMCS)-modified β-cyclodextrin fisetin nanoparticle (β-CD-Fis-CMCS).
ModelMale C57BL/6J mice (8 weeks old) fed a high-fat, high-cholesterol (HFHC) diet for 12 weeks; murine AML12 hepatocyte cell line challenged with cholesterol and 25-hydroxycholesterol; ASGR1-overexpression plasmid transfection in AML12 cells.
SampleMouse study: n=8 per group across eight groups (64 mice total). In vitro assays: n=3 per condition.
InterventionOral fisetin at 12.5 or 25 mg/kg/day; simvastatin 5 mg/kg/day as active comparator; β-CD-Fis-CMCS nanoparticles delivering 5 or 10 mg/kg/day fisetin-equivalent dose, all administered for 8 weeks after 4 weeks of HFHC diet induction (12 weeks total diet, mice aged 8 weeks at start); in vitro fisetin 0.5-2 µg/mL or equivalent β-CD-Fis-CMCS nanoparticle concentrations (as low as one-fifth the free-fisetin dose).
Duration12-week total HFHC diet protocol with 8 weeks of concurrent fisetin/nanoparticle/simvastatin administration; in vitro treatments up to 48 hours.
EndpointsHepatic lipid accumulation (Oil Red O staining); Serum total cholesterol, triglycerides, LDL-C and HDL-C; Oxidative stress markers (serum MDA, SOD); Hepatic cholesterol-metabolism gene/protein expression (ABCA1, ABCG5, ABCG8, LDLR, SR-B1, LXRα, HMGCR, CYP7A1); ASGR1 expression and molecular docking/CETSA target engagement; mTORC1/AMPK-BRCA1/BARD1 pathway phosphorylation status; Nanoparticle physicochemical characterisation (particle size, zeta potential, encapsulation efficiency, drug loading, release kinetics) and in vivo biodistribution

What the study showed, in plain terms

This very recent 2026 Chinese study asks whether fisetin can help lower cholesterol, and if so, through what mechanism — using a mouse model of diet-induced high cholesterol (hypercholesterolemia), a known driver of heart-disease risk.

Mice fed a high-fat, high-cholesterol diet for 12 weeks developed fatty liver and elevated blood cholesterol and triglycerides. Adding oral fisetin (12.5-25 mg/kg/day) for 8 of those weeks reduced liver fat accumulation by roughly a quarter to three-quarters (depending on dose) and lowered blood total cholesterol, triglycerides and LDL ('bad') cholesterol, working about as well as a low dose of the statin drug simvastatin used for comparison.

The mechanistic story is genuinely novel: the researchers found fisetin binds to a liver-cell surface receptor called ASGR1 (asialoglycoprotein receptor 1), which normally helps clear proteins from the blood but has recently been linked to cholesterol regulation. By reducing ASGR1 activity, fisetin shifted a cellular energy-sensing pathway (mTORC1/AMPK) in a way that activated a DNA-repair-protein complex (BRCA1/BARD1) and ultimately turned up several cholesterol-clearing pathways in the liver — more bile acid production, more cholesterol pumped out via transporters called ABCA1/ABCG5/ABCG8, and reduced cholesterol synthesis.

To address fisetin's well-known solubility problem, the team also built a nanoparticle version (fisetin wrapped in a modified cyclodextrin/chitosan shell) and showed it worked just as well, or better, at only one-fifth the dose of plain fisetin — directly relevant to the Data Center's broader interest in fisetin delivery technology. As with all mouse studies, this cannot be assumed to translate directly to a human cholesterol-lowering effect, and this is a single very recent paper (published February 2026) that has not yet been replicated by other groups.

Key findings

  • Oral fisetin (12.5 or 25 mg/kg/day, 8 weeks) reduced hepatic lipid accumulation by approximately 28% and 77% respectively (Oil Red O staining) in HFHC diet-fed mice, and significantly lowered serum total cholesterol, triglycerides and LDL-C without significantly changing HDL-C.
  • Fisetin reversed HFHC-induced oxidative stress, reducing serum MDA by roughly 44% (relative to model) and restoring SOD activity, roughly matching the effect of simvastatin.
  • Fisetin modulated the reverse cholesterol transport pathway, upregulating hepatic ABCA1, LDLR and SR-B1 protein/gene expression (by 282%, 248% and 142% respectively at 25 mg/kg vs model) and restoring LXRα, ABCG5, ABCG8 and CYP7A1 while lowering the cholesterol-synthesis enzyme HMGCR.
  • Molecular docking (binding energy approximately -7.0 to -7.1 kcal/mol) and cellular thermal shift assay (CETSA) confirmed fisetin directly binds asialoglycoprotein receptor 1 (ASGR1); fisetin reduced HFHC/HC-induced ASGR1 upregulation in both mouse liver and AML12 hepatocytes.
  • Fisetin reduced ASGR1 co-localisation with the lysosomal marker CD63, and modulated the downstream mTORC1/AMPK-BRCA1/BARD1 pathway (decreased phosphorylated mTOR and S6K, increased phosphorylated AMPK, decreased BRCA1/BARD1), consistent with enhanced cholesterol efflux signalling.
  • Overexpressing ASGR1 in AML12 cells reversed fisetin's cholesterol-lowering and pathway effects, confirming ASGR1 as fisetin's functional target rather than an incidental correlation.
  • A carboxymethyl chitosan-modified β-cyclodextrin fisetin nanoparticle (β-CD-Fis-CMCS, ~40 nm diameter) achieved comparable or superior cholesterol-lowering and lipid-accumulation effects at one-fifth the fisetin dose of the free compound, both in AML12 cells and in HFHC diet-fed mice, and showed liver-targeted biodistribution in vivo.

What this study can and cannot tell us

This is a single, very recently published (February 2026) mouse and cell-culture study; it has not yet been independently replicated, and the authors themselves note that mice lack the cholesterol ester transfer protein (CETP) that plays an important role in human lipid metabolism, which may limit direct translation of these findings to humans.

The nanoparticle formulation (β-CD-Fis-CMCS) is a novel laboratory research prototype, not a tested or marketed supplement product, and its in vitro release testing used simulated colonic fluid only, not a fuller simulation of gastric and small-intestinal conditions.

The paper notes as its own limitation that the fisetin nanoparticles lack an active liver-targeting ligand (such as GalNAc, which binds ASGR1 with high affinity) — a refinement the authors suggest as a future direction that could further improve hepatic delivery.

Oral gavage doses in mice (12.5-25 mg/kg/day) are, as with other fisetin animal studies in this Data Center, higher on a body-weight basis than typical human oral supplement doses, and cannot be assumed to produce an equivalent cholesterol-lowering effect in people.

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Dr. Khaled Mahmoud, MBBChCritical Care Medicine, Longevity Science & Editorial Direction Dr. Amr Yasser, PhDCardiometabolic, Musculoskeletal, Gut & Bioavailability