Tier 4 — mechanistic

Fisetin limits Chikungunya virus-induced apoptosis hallmarks in hepatocellular carcinoma cells

Lani R, Hassandarvish P, AbuBakar S
Scientific Reports 2025 Volume 15, article 25140

Bibliography

PubMed
PMID 40645991
PubMed Central
PMC12254361
DOI
10.1038/s41598-025-09213-6
Funding
Ministry of Higher Education Malaysia, Higher Institution Centre of Excellence (HICoE) program (grant nos. MO002-2019 and TIDREC-2023) and SUKUK PRIHATIN-Fasa 2 grant MO002-2021.
Competing interests
The authors declare no competing interests.

Study snapshot

DesignIn vitro antiviral/cytoprotection study using plaque-forming unit assays, qPCR viral load quantification, DNA fragmentation assays, immunoblotting and a proteome profiler apoptosis array.
ModelHuman hepatocellular carcinoma cell line Huh7 infected with Chikungunya virus (East/Central/South African genotype, MOI = 1).
SampleStandard in vitro replicate design typical of this laboratory's prior published CHIKV/fisetin work; specific per-assay replicate counts not extracted from the available pages.
InterventionFisetin 30 µM added post-CHIKV infection, benchmarked against Z-VAD-FMK (20 µM, pan-caspase inhibitor) and cisplatin (130 µM, pro-apoptotic positive control).
DurationViral and apoptosis endpoints assessed at 24 and 48 hours post-infection.
EndpointsInfectious viral particle count (plaque-forming unit assay); CHIKV RNA levels (qPCR, E1 gene, supernatant and intracellular); DNA fragmentation (laddering assay); Cleaved PARP (immunoblot); Apoptosis-related proteins: cleaved caspase-3, HSP-27, HIF-1α (Proteome Profiler Human Apoptosis Array)

What the study showed, in plain terms

This Malaysian research group had previously shown fisetin can inhibit Chikungunya virus replication in cells; this newer paper dug into what fisetin does to virus-infected liver cancer cells at the level of programmed cell death (apoptosis), using human hepatocellular carcinoma (Huh7) cells as the infection model.

Fisetin significantly reduced both the amount of Chikungunya virus RNA and the number of infectious virus particles produced by infected cells, and it did this more effectively than a standard caspase-blocking drug (Z-VAD-FMK) or cisplatin, a chemotherapy drug used here as a comparison for triggering apoptosis.

On the cell-death side, fisetin produced the least DNA fragmentation (a hallmark of apoptosis) among all the treatments tested, prevented the classic cleavage of a protein called PARP that signals apoptosis is underway, and reduced levels of activated caspase-3 and a stress protein called HSP-27, while restoring a protein (HIF-1α) that the virus had suppressed.

Put together, the findings suggest fisetin has a dual action here: it fights the virus directly (antiviral) while also protecting the infected liver cells from dying in the particular apoptotic way the virus triggers (cytoprotective) — a combination the authors say is a novel finding for this specific virus-cell combination, though they are careful to note fisetin's well-known poor absorption and rapid breakdown in the body as a real hurdle before this could matter clinically.

Key findings

  • Fisetin (30 µM) significantly reduced CHIKV RNA levels (supernatant and intracellular) and infectious viral particle counts at 24 and 48 hours post-infection (p<0.0001), outperforming both Z-VAD-FMK and cisplatin.
  • Fisetin produced the lowest level of DNA fragmentation among all treatment groups, indicating the strongest anti-apoptotic effect against CHIKV-induced cell death.
  • No cleaved PARP was detected in the CHIKV-plus-fisetin group, in contrast to strong PARP cleavage in the CHIKV-plus-cisplatin group, indicating fisetin blocked this apoptotic signalling step.
  • Fisetin significantly reduced cleaved caspase-3 and HSP-27 levels (both elevated by CHIKV infection) and restored HIF-1α protein levels (reduced by CHIKV infection), all p<0.0001 versus untreated infected cells.
  • The authors note this is the first study to show fisetin modulates CHIKV-induced apoptotic hallmarks specifically in hepatocytes, suggesting a dual antiviral and cytoprotective role, building on the same group's earlier work establishing fisetin's IC50 of 20-35 µM against CHIKV with no cytotoxicity up to 1.22 mM.

What this study can and cannot tell us

This is a single-cell-line in vitro study (Huh7 hepatocellular carcinoma cells); there is no animal model of Chikungunya infection or human clinical data to confirm antiviral or cytoprotective efficacy in vivo.

The authors themselves flag fisetin's poor oral bioavailability and rapid metabolism as a translational limitation, meaning the 30 µM concentration used in vitro may not be readily achievable in human plasma with standard oral fisetin supplementation.

Huh7 is a hepatocellular carcinoma (cancer) cell line, not normal primary human hepatocytes, so results may not fully generalise to how fisetin would behave in non-cancerous liver tissue during a real Chikungunya infection.

Editorial review

Reviewed by the Biohack Blueprint research team

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Dr. Khaled Mahmoud, MBBChCritical Care Medicine, Longevity Science & Editorial Direction Dr. Amr Yasser, PhDCardiometabolic, Musculoskeletal, Gut & Bioavailability Dr. Abdulla Mostafa, PhDCellular Senescence, Neurological, Mitochondrial & Brain-Ageing Biology