J Food Bioact

Journal of Food Bioactives, ISSN 2637-8752 print, 2637-8779 online

Journal website www.isnff-jfb.com

Review

Volume 31, September 2025, pages 41-62

Beyond petri dish: small animal models bridge in vitro and in vivo antioxidant assays

Figure 1. Different antioxidant detection methods using zebrafish models.

Figure 2. D. melanogaster as a small in vivo model to study antioxidant activities of food derived antioxidants.

Figure 3. Overview of Caenorhabditis elegans as an antioxidant research model

**Table 1.** Use of Zebrafish, Fruit Fly, and *C. elegans* for Antioxidant Discovery
Compound/Extract	Source	Key Findings in Zebrafish	Mechanisms/Pathways	References
Zebra fish model
Water Extract of Citrus Pomace	Citrus Pomace	Improved survival rates; reduced heartbeat and ROS levels.	Scavenging of free radicals; reduction of cell death.	Wang et al., 2018
Heat-killed LAB Strains	Various Lactic Acid Bacteria	Reduced H₂O₂-induced toxicity; all strains showed antioxidant activity.	Indirect antioxidant activity; activation of antioxidant enzymes.	Sato et al., 2024
Rutin Trihydrate	Not specified	Increased GPx (9%), GSH (5%), GST (5%); countered doxorubicin effects.	Upregulation of key antioxidant enzymes; oxidative stress response.	Seethalakshmi and Kumar, 2024
6-Gingerol	Ginger	Reduced ROS and cell death; ↑SOD and CAT; ↓lipid peroxidation in larvae.	Direct antioxidant activity; regulation of ROS and H₂O₂.	Manjunathan et al., 2021
Naringenin	Citrus, food sources	Protective against oxidative stress-induced lethality.	Induction of antioxidant pathways (e.g., Nrf2).	Arteaga et al., 2021
Apigenin	Parsley, food sources	Protective against lethality and dysmorphogenesis.	Activation of cellular protective responses.	Arteaga et al., 2021
Rutin	Buckwheat, food sources	Protective against lethality and dysmorphogenesis.	Enhanced oxidative stress resistance.	Arteaga et al., 2021
Oleuropein	Olive, food sources	Protective against oxidative stress-induced lethality.	Modulation of oxidative stress mechanisms.	Arteaga et al., 2021
Chlorogenic Acid	Coffee, food sources	Protective against oxidative stress-induced lethality.	Support of antioxidant defense systems.	Arteaga et al., 2021
Curcumin	Turmeric, food sources	Protective against lethality and dysmorphogenesis.	Activation of Nrf2 and other protective pathways.	Arteaga et al., 2021
Lycopene	Tomato, food sources	Protective against oxidative stress-induced lethality.	Enhancement of antioxidant pathways.	Arteaga et al., 2021
Astaxanthin	Algae, food sources	Protective against oxidative stress-induced lethality.	Modulation of cellular oxidative stress response.	Arteaga et al., 2021
β-Carotene	Carrot, food sources	Increased lethality and dysmorphogenesis at tested conditions.	Potential pro-oxidant effects at certain concentrations.	Arteaga et al., 2021
Carnosine	–	Decreased ROS in larvae; rescued Hsp70 and MT expression altered by TiO₂ NPs.	Antioxidant activity counteracting TiO₂ nanoparticle-induced oxidative stress.	Caruso et al., 2023
Phosvitin	Zebrafish Egg Yolk	Effective antioxidant; inhibited linoleic acid oxidation; non-cytotoxic.	Scavenges DPPH radicals; protects biomolecules from oxidation.	Hu et al., 2015
Chlorogenic Acid	Sonchus oleraceus (Sowthistle)	Reduced phenotypic abnormalities in embryos exposed to pro-oxidant auranofin.	Protective effects against oxidative stress and malformation.	Chiu et al., 2020
Nrf2 Inducers	Various Compounds	Induced expression of antioxidant genes (HO-1, NQO1, GSTs).	Activation of the Nrf2-ARE pathway.	Jung and Kwak, 2010
Quercetin Nanocrystals	Nanosuspension	Enhanced survival rates (66.67–77.78%) in H₂O₂-treated zebrafish.	Reduced ROS levels; catalyzed increased CAT and GPx activity.	Wang et al., 2023a
Glutathione (GSH)	Endogenous System	Robust antioxidant system; oxidized by ROS forming GSSG.	Involves GPx and GR; key oxidative stress marker.	Massarsky et al., 2017
N/A (Review on Drug Screening)	N/A	Review of zebrafish as a model for novel anti-inflammatory drug screening.	Analysis of inflammatory response and leukocyte behavior.	Xie et al., 2021
Lignin-Carbohydrate Complexes	Wheat Stalk	Protected against BPA-induced neurotoxicity; reduced ROS and neuronal damage.	Suppressed nerve-related gene expression; reduced oxidative stress.	Gu et al., 2021
Polyphenolic Extract	Condalia microphylla Fruits	Inhibited lipid oxidation by up to 40% under oxidative stress.	Antioxidant activity; reduction of oxidative stress.	Boeri et al., 2017
Catechol	Experimental Compound	Increased ROS, reduced antioxidant capacity; decreased locomotor activity, increased anxiety.	Induction of redox imbalance; behavioral disruption.	Xiao et al., 2025
Equol	Soy	Exerts strong antioxidant effects.	Functions through an Nrf2-independent mechanism.	Watanabe et al., 2022
Total Particulate Matter (TPM)	Cigarette Smoke	Increased mortality, delayed hatching, deformities, and behavioral changes.	Disrupted angiogenesis; affected xenobiotic metabolism and oxidative stress.	Massarsky et al., 2015
tBOOH	Oxidant (Positive Control)	Induces oxidative stress in zebrafish embryos.	Used as a model oxidant to test protective effects of antioxidants.	Boix et al., 2020
TCHQ	Oxidant (Positive Control)	Induces oxidative stress in zebrafish embryos.	Used as a model oxidant to test protective effects of antioxidants.	Boix et al., 2020
LPS	Escherichia coli	Induces oxidative stress in zebrafish embryos.	Used to model inflammatory/oxidative stress to test antioxidants.	Boix et al., 2020
Momordica cochinchinensis Extract	Gac Fruit	Improved locomotor functions in an MPTP-induced Parkinson’s disease model.	Not linked to significant change in antioxidant enzyme activities.	Singsai et al., 2023
Formulated Agar 1	Turbinaria conoides (Seaweed)	Higher antioxidant activity; lower teratogenic effects.	Demonstrates dose-dependent antioxidant potential.	Aavula et al., 2022
Formulated Agar 2	Turbinaria conoides (Seaweed)	Lower antioxidant activity; higher teratogenic effects.	Demonstrates dose-dependent toxicity.	Aavula et al., 2022
Curcumin	Turmeric	Reduces hydrogen peroxide-induced toxicity.	Nrf2-dependent pathway activation.	Endo et al., 2020
Diallyl Trisulfide	Garlic	Reduces hydrogen peroxide-induced toxicity.	Nrf2-dependent pathway activation.	Endo et al., 2020
Quercetin	Various plants	Reduces hydrogen peroxide-induced toxicity.	Nrf2-dependent pathway activation.	Endo et al., 2020
Cinnamaldehyde	Cinnamon	Reduces arsenite toxicity; exhibits antioxidant effects.	Nrf2-independent pathway.	Endo et al., 2020
Carnosic Acid	Rosemary	Toxic at high concentrations; antioxidant analysis inconclusive.	Proposed Nrf2 pathway; confounded by toxicity.	Endo et al., 2020
Troxerutin	Semi-synthetic flavonoid	Reduced MPO, NO, and LPO; protective against nicotine-induced lung fibrosis.	Scavenges free radicals; enhances defense; suppresses IL-10 & IL-1β expression.	Hobani, 2024
Dieckol	Ecklonia cava (Brown seaweed)	Reduced ROS, nitric oxide, and cell death induced by UVB radiation.	Direct antioxidant activity; protective against UVB-induced damage.	Wang et al., 2021
Antioxidant Peptides	C-phycocyanin (from algae)	Protective against H₂O₂-induced oxidative stress.	Activation of Nrf2 signaling pathway.	Xu et al., 2022
Lignin–carbohydrate complexes (LCCs)	Bamboo and Poplar	Scavenged endogenous ROS effectively.	Prevented reduction of antioxidant enzyme activity (SOD, CAT).	Dong et al., 2019
Fruit fly model
Cyanidin-3-O-glucoside (Anthocyanin)	Lonicera pallasii (Honeysuckle)	Increased lifespan and stress resistance; improved intestinal barrier integrity	Sirt6 activation; regulation of Hif1 & Keap1	Golubev et al., 2022
Mangifera indica leaf extract	Mangifera indica (Mango)	Enhanced survival; ↑GST, CAT activity; ↑thiol content	Enzyme induction (dose-sensitive)	Alexander et al., 2019
Astragalus membranaceus extract (AME)	Astragalus membranaceus	Enhanced survival under H₂O₂ challenge; ↓ROS; ↑SOD, CAT; ↑Sod1, Cat, CncC	Enzyme induction; gene regulation	Dai et al., 2024
Curcumin	Turmeric	Lifespan extension (males); reversed by SOD inhibitor	SOD-dependent pathway	Suckow and Suckow, 2006
Flavonoids (icariin, epimedins A–C)	Epimedium pubescens	Enhanced radical-scavenging; boosted CAT, GSH-Px	Antioxidant enzyme activation	Yang et al., 2020
Proanthocyanidin-rich fraction	Tamarindus indica	Improved lifespan, emergence rate, antioxidant enzyme activity; ↓AChE, caspase-3/9	Antioxidant enzyme activation; neuroprotection	Jaafaru et al., 2024
Artemisia argyi extract	Artemisia argyi	Prolonged lifespan; improved climbing; ↑stress tolerance; enzyme modulation; ↓MDA	SOD, CAT modulation	Yang et al., 2024
Phlorizin	Apple	Lifespan extension; improved locomotion; ↑Nrf2/cnc, ↓methuselah	Nrf2 activator; stress regulation	Wang et al., 2019
Crimson snapper scale peptides	Marine fish	Extended lifespan; ↓MDA, protein carbonylation; ↑SOD, CAT	Upregulation of SOD1, SOD2, CAT	Chen et al., 2020
Rice protein hydrolysates	Rice	Increased lifespan; boosted SOD, Mn-SOD, CAT	Nrf2/Keap1, TOR/S6K, methuselah modulation	Yue et al., 2021
Casein peptides	Milk protein	Improved survival; restored GSH, thiols, proteins; normalized oxidative markers	Keap1/Nrf2 regulation	Sadiq et al., 2023
Lateolabrax japonicus peptides (LPH)	Japanese seabass	Extended lifespan; ↓ROS, MDA; ↑SOD, CAT, GSH-Px; preserved gut integrity	Nrf2 activation; mTOR downregulation; gut microbiota modulation	Li et al., 2023b
C. elegans model
Rhein Derivative 4b	Rhein	Increases lifespan & stress resistance; enhances GSH; reduces MDA & ROS	Targets Keap1-Nrf2 pathway	Wang, 2025
Monascus-fermented Dioscorea (RMDE)	Yam	Increases survival during oxidative stress; reduces ROS	DAF-16/FOXO-dependent pathway; induces sod-3 expression	Shi et al., 2012
Magnolol Derivative M27	Houpoea officinalis	Prolongs lifespan; improves healthspan; increases stress resistance	IIS pathway (DAF-2, AGE-1, DAF-16)	Pang et al., 2023
Polyphenols	Blumea laciniata	Extends lifespan (17.4%); enhances stress resistance; reduces ROS & MDA	Insulin/IGF-1 signaling; promotes DAF-16 nuclear translocation	Chen et al., 2021
Caffeic Acid Phenethylester (CAPE)	Propolis	Increases stress resistance & lifespan (∼9–17%); reduces ROS	Modulates DAF-16 signaling pathway; SKN-1 independent	Havermann et al., 2014
Methanol Extract	Camellia tenuifolia	Prolongs lifespan; reduces amyloid-β toxicity	Decreases intracellular ROS levels	Wei et al., 2014
Kaempferol Glycosides	Camellia tenuifolia	Decreases ROS levels; prolongs lifespan	Antioxidant and anti-inflammatory activities	Wei et al., 2014
Hot Water Extract	Chamaecyparis obtusa	Extends lifespan; decreases lipofuscin	Enhanced antioxidant activity via flavonoid absorption	Cheng et al., 2014
Guarana	Paullinia cupana	Extends lifespan; antioxidant activity	DAF-16, HSF-1, SKN-1 pathways	Arantes et al. 2018
Selenite	–	Protects from oxidative stress	DAF-16 and TRXR-1 dependent	Li et al., 2013
Oleuropein	Olive	Prolongs lifespan (22.3%); increases survival against stress	Insulin/IGF-1 and SKN-1/Nrf2 signaling pathways	Fang and Miller, 2012
Yeast Hydrolysate	Yeast	Enhances lifespan and stress resistance	Modulates tyrosine, glycerophospholipid, and glutathione metabolism	Li et al., 2023a
Guarana	Paullinia cupana	Lifespan extension; high antioxidant capacity	Indirect effects via altering E. coli metabolism	Reigada et al., 2022
Phenolic Compounds	Sonchus arvensis, Hemerocallis citrina	Improves lifespan & health against nutrient stress	TGF-β signaling pathway; skn-1	An et al., 2023
Cassia fistula Extract	Cassia fistula	Improves survival; reduces ROS & proteotoxicity	DAF-16/FOXO and SKN-1/NRF2 pathways	Thabit et al., 2018
Carbendazim	Fungicide	Inhibits growth & lifespan; damages reproduction & antioxidant systems	Impairs oxidative stress response; toxic	Li et al., 2020
Liuwei Dihuang (LWDH)	Traditional Chinese Medicine	Delays β-amyloid paralysis; reduces ROS	Upregulation of HSPs; antioxidant activity	Sangha et al., 2012
Ethyl Acetate Extract	Gastrodia elata	Protects from oxidative stress & Aβ toxicity; improves lifespan	Insulin/IGF-1 signaling (IIS) pathway	Shi et al., 2023
Acrolein	Toxic Aldehyde	Shortens lifespan; increases ROS; decreases healthspan	Activates DAF-16/FOXO stress response	Jeayeng et al., 2024
Leaf Extract	Anacardium occidentale (Cashew)	Enhances stress survival & lifespan; reduces lipofuscin	DAF-16/FoxO & SKN-1/Nrf-2 pathways; induces sod-3, gst-4	Duangjan et al., 2019
Polysaccharides	Dendrobium officinale	Prolongs lifespan; increases antioxidant enzyme activity	Upregulation of daf-16, skn-1, sir-2.1	Tang et al., 2023
Total Flavonoids	Sea Buckthorn	Increases lifespan (29.4%); enhances stress tolerance; delays paralysis	Radical scavenging; AChE/MAO-A inhibition	Wang et al., 2022
Polysaccharides	Fermented Coix Seed	Increases lifespan (5.9%); enhances antioxidant enzymes	Downregulates daf-2, age-1; upregulates daf-16, sod-3, skn-1	Zhao et al., 2023
Cannabidiol (CBD)	Cannabis sativa	Extends lifespan & survival in AD model	Activates neural glyoxalase pathway; detoxifies methylglyoxal	Frandsen and Narayanasamy, 2022
Extract	Anoectochilus roxburghii	Prolongs lifespan; reduces ROS; increases stress resistance	Activation of daf-16/FoxO pathway	Xu et al., 2024

Table 1. Use of Zebrafish, Fruit Fly, and C. elegans for Antioxidant Discovery

Compound/Extract

Source

Key Findings in Zebrafish

Mechanisms/Pathways

References

Zebra fish model

Water Extract of Citrus Pomace

Citrus Pomace

Improved survival rates; reduced heartbeat and ROS levels.

Scavenging of free radicals; reduction of cell death.

Wang et al., 2018

Heat-killed LAB Strains

Various Lactic Acid Bacteria

Reduced H₂O₂-induced toxicity; all strains showed antioxidant activity.

Indirect antioxidant activity; activation of antioxidant enzymes.

Sato et al., 2024

Rutin Trihydrate

Not specified

Increased GPx (9%), GSH (5%), GST (5%); countered doxorubicin effects.

Upregulation of key antioxidant enzymes; oxidative stress response.

Seethalakshmi and Kumar, 2024

6-Gingerol

Ginger

Reduced ROS and cell death; ↑SOD and CAT; ↓lipid peroxidation in larvae.

Direct antioxidant activity; regulation of ROS and H₂O₂.

Manjunathan et al., 2021

Naringenin

Citrus, food sources

Protective against oxidative stress-induced lethality.

Induction of antioxidant pathways (e.g., Nrf2).

Arteaga et al., 2021

Apigenin

Parsley, food sources

Protective against lethality and dysmorphogenesis.

Activation of cellular protective responses.

Arteaga et al., 2021

Rutin

Buckwheat, food sources

Protective against lethality and dysmorphogenesis.

Enhanced oxidative stress resistance.

Arteaga et al., 2021

Oleuropein

Olive, food sources

Protective against oxidative stress-induced lethality.

Modulation of oxidative stress mechanisms.

Arteaga et al., 2021

Chlorogenic Acid

Coffee, food sources

Protective against oxidative stress-induced lethality.

Support of antioxidant defense systems.

Arteaga et al., 2021

Curcumin

Turmeric, food sources

Protective against lethality and dysmorphogenesis.

Activation of Nrf2 and other protective pathways.

Arteaga et al., 2021

Lycopene

Tomato, food sources

Protective against oxidative stress-induced lethality.

Enhancement of antioxidant pathways.

Arteaga et al., 2021

Astaxanthin

Algae, food sources

Protective against oxidative stress-induced lethality.

Modulation of cellular oxidative stress response.

Arteaga et al., 2021

β-Carotene

Carrot, food sources

Increased lethality and dysmorphogenesis at tested conditions.

Potential pro-oxidant effects at certain concentrations.

Arteaga et al., 2021

Carnosine

–

Decreased ROS in larvae; rescued Hsp70 and MT expression altered by TiO₂ NPs.

Antioxidant activity counteracting TiO₂ nanoparticle-induced oxidative stress.

Caruso et al., 2023

Phosvitin

Zebrafish Egg Yolk

Effective antioxidant; inhibited linoleic acid oxidation; non-cytotoxic.

Scavenges DPPH radicals; protects biomolecules from oxidation.

Hu et al., 2015

Chlorogenic Acid

Sonchus oleraceus (Sowthistle)

Reduced phenotypic abnormalities in embryos exposed to pro-oxidant auranofin.

Protective effects against oxidative stress and malformation.

Chiu et al., 2020

Nrf2 Inducers

Various Compounds

Induced expression of antioxidant genes (HO-1, NQO1, GSTs).

Activation of the Nrf2-ARE pathway.

Jung and Kwak, 2010

Quercetin Nanocrystals

Nanosuspension

Enhanced survival rates (66.67–77.78%) in H₂O₂-treated zebrafish.

Reduced ROS levels; catalyzed increased CAT and GPx activity.

Wang et al., 2023a

Glutathione (GSH)

Endogenous System

Robust antioxidant system; oxidized by ROS forming GSSG.

Involves GPx and GR; key oxidative stress marker.

Massarsky et al., 2017

N/A (Review on Drug Screening)

N/A

Review of zebrafish as a model for novel anti-inflammatory drug screening.

Analysis of inflammatory response and leukocyte behavior.

Xie et al., 2021

Lignin-Carbohydrate Complexes

Wheat Stalk

Protected against BPA-induced neurotoxicity; reduced ROS and neuronal damage.

Suppressed nerve-related gene expression; reduced oxidative stress.

Gu et al., 2021

Polyphenolic Extract

Condalia microphylla Fruits

Inhibited lipid oxidation by up to 40% under oxidative stress.

Antioxidant activity; reduction of oxidative stress.

Boeri et al., 2017

Catechol

Experimental Compound

Increased ROS, reduced antioxidant capacity; decreased locomotor activity, increased anxiety.

Induction of redox imbalance; behavioral disruption.

Xiao et al., 2025

Equol

Soy

Exerts strong antioxidant effects.

Functions through an Nrf2-independent mechanism.

Watanabe et al., 2022

Total Particulate Matter (TPM)

Cigarette Smoke

Increased mortality, delayed hatching, deformities, and behavioral changes.

Disrupted angiogenesis; affected xenobiotic metabolism and oxidative stress.

Massarsky et al., 2015

tBOOH

Oxidant (Positive Control)

Induces oxidative stress in zebrafish embryos.

Used as a model oxidant to test protective effects of antioxidants.

Boix et al., 2020

TCHQ

Oxidant (Positive Control)

Induces oxidative stress in zebrafish embryos.

Used as a model oxidant to test protective effects of antioxidants.

Boix et al., 2020

LPS

Escherichia coli

Induces oxidative stress in zebrafish embryos.

Used to model inflammatory/oxidative stress to test antioxidants.

Boix et al., 2020

Momordica cochinchinensis Extract

Gac Fruit

Improved locomotor functions in an MPTP-induced Parkinson’s disease model.

Not linked to significant change in antioxidant enzyme activities.

Singsai et al., 2023

Formulated Agar 1

Turbinaria conoides (Seaweed)

Higher antioxidant activity; lower teratogenic effects.

Demonstrates dose-dependent antioxidant potential.

Aavula et al., 2022

Formulated Agar 2

Turbinaria conoides (Seaweed)

Lower antioxidant activity; higher teratogenic effects.

Demonstrates dose-dependent toxicity.

Aavula et al., 2022

Curcumin

Turmeric

Reduces hydrogen peroxide-induced toxicity.

Nrf2-dependent pathway activation.

Endo et al., 2020

Diallyl Trisulfide

Garlic

Reduces hydrogen peroxide-induced toxicity.

Nrf2-dependent pathway activation.

Endo et al., 2020

Quercetin

Various plants

Reduces hydrogen peroxide-induced toxicity.

Nrf2-dependent pathway activation.

Endo et al., 2020

Cinnamaldehyde

Cinnamon

Reduces arsenite toxicity; exhibits antioxidant effects.

Nrf2-independent pathway.

Endo et al., 2020

Carnosic Acid

Rosemary

Toxic at high concentrations; antioxidant analysis inconclusive.

Proposed Nrf2 pathway; confounded by toxicity.

Endo et al., 2020

Troxerutin

Semi-synthetic flavonoid

Reduced MPO, NO, and LPO; protective against nicotine-induced lung fibrosis.

Scavenges free radicals; enhances defense; suppresses IL-10 & IL-1β expression.

Hobani, 2024

Dieckol

Ecklonia cava (Brown seaweed)

Reduced ROS, nitric oxide, and cell death induced by UVB radiation.

Direct antioxidant activity; protective against UVB-induced damage.

Wang et al., 2021

Antioxidant Peptides

C-phycocyanin (from algae)

Protective against H₂O₂-induced oxidative stress.

Activation of Nrf2 signaling pathway.

Xu et al., 2022

Lignin–carbohydrate complexes (LCCs)

Bamboo and Poplar

Scavenged endogenous ROS effectively.

Prevented reduction of antioxidant enzyme activity (SOD, CAT).

Dong et al., 2019

Fruit fly model

Cyanidin-3-O-glucoside (Anthocyanin)

Lonicera pallasii (Honeysuckle)

Increased lifespan and stress resistance; improved intestinal barrier integrity

Sirt6 activation; regulation of Hif1 & Keap1

Golubev et al., 2022

Mangifera indica leaf extract

Mangifera indica (Mango)

Enhanced survival; ↑GST, CAT activity; ↑thiol content

Enzyme induction (dose-sensitive)

Alexander et al., 2019

Astragalus membranaceus extract (AME)

Astragalus membranaceus

Enhanced survival under H₂O₂ challenge; ↓ROS; ↑SOD, CAT; ↑Sod1, Cat, CncC

Enzyme induction; gene regulation

Dai et al., 2024

Curcumin

Turmeric

Lifespan extension (males); reversed by SOD inhibitor

SOD-dependent pathway

Suckow and Suckow, 2006

Flavonoids (icariin, epimedins A–C)

Epimedium pubescens

Enhanced radical-scavenging; boosted CAT, GSH-Px

Antioxidant enzyme activation

Yang et al., 2020

Proanthocyanidin-rich fraction

Tamarindus indica

Improved lifespan, emergence rate, antioxidant enzyme activity; ↓AChE, caspase-3/9

Antioxidant enzyme activation; neuroprotection

Jaafaru et al., 2024

Artemisia argyi extract

Artemisia argyi

Prolonged lifespan; improved climbing; ↑stress tolerance; enzyme modulation; ↓MDA

SOD, CAT modulation

Yang et al., 2024

Phlorizin

Apple

Lifespan extension; improved locomotion; ↑Nrf2/cnc, ↓methuselah

Nrf2 activator; stress regulation

Wang et al., 2019

Crimson snapper scale peptides

Marine fish

Extended lifespan; ↓MDA, protein carbonylation; ↑SOD, CAT

Upregulation of SOD1, SOD2, CAT

Chen et al., 2020

Rice protein hydrolysates

Rice

Increased lifespan; boosted SOD, Mn-SOD, CAT

Nrf2/Keap1, TOR/S6K, methuselah modulation

Yue et al., 2021

Casein peptides

Milk protein

Improved survival; restored GSH, thiols, proteins; normalized oxidative markers

Keap1/Nrf2 regulation

Sadiq et al., 2023

Lateolabrax japonicus peptides (LPH)

Japanese seabass

Extended lifespan; ↓ROS, MDA; ↑SOD, CAT, GSH-Px; preserved gut integrity

Nrf2 activation; mTOR downregulation; gut microbiota modulation

Li et al., 2023b

C. elegans model

Rhein Derivative 4b

Rhein

Increases lifespan & stress resistance; enhances GSH; reduces MDA & ROS

Targets Keap1-Nrf2 pathway

Wang, 2025

Monascus-fermented Dioscorea (RMDE)

Yam

Increases survival during oxidative stress; reduces ROS

DAF-16/FOXO-dependent pathway; induces sod-3 expression

Shi et al., 2012

Magnolol Derivative M27

Houpoea officinalis

Prolongs lifespan; improves healthspan; increases stress resistance

IIS pathway (DAF-2, AGE-1, DAF-16)

Pang et al., 2023

Polyphenols

Blumea laciniata

Extends lifespan (17.4%); enhances stress resistance; reduces ROS & MDA

Insulin/IGF-1 signaling; promotes DAF-16 nuclear translocation

Chen et al., 2021

Caffeic Acid Phenethylester (CAPE)

Propolis

Increases stress resistance & lifespan (∼9–17%); reduces ROS

Modulates DAF-16 signaling pathway; SKN-1 independent

Havermann et al., 2014

Methanol Extract

Camellia tenuifolia

Prolongs lifespan; reduces amyloid-β toxicity

Decreases intracellular ROS levels

Wei et al., 2014

Kaempferol Glycosides

Camellia tenuifolia

Decreases ROS levels; prolongs lifespan

Antioxidant and anti-inflammatory activities

Wei et al., 2014

Hot Water Extract

Chamaecyparis obtusa

Extends lifespan; decreases lipofuscin

Enhanced antioxidant activity via flavonoid absorption

Cheng et al., 2014

Guarana

Paullinia cupana

Extends lifespan; antioxidant activity

DAF-16, HSF-1, SKN-1 pathways

Arantes et al. 2018

Selenite

–

Protects from oxidative stress

DAF-16 and TRXR-1 dependent

Li et al., 2013

Oleuropein

Olive

Prolongs lifespan (22.3%); increases survival against stress

Insulin/IGF-1 and SKN-1/Nrf2 signaling pathways

Fang and Miller, 2012

Yeast Hydrolysate

Yeast

Enhances lifespan and stress resistance

Modulates tyrosine, glycerophospholipid, and glutathione metabolism

Li et al., 2023a

Guarana

Paullinia cupana

Lifespan extension; high antioxidant capacity

Indirect effects via altering E. coli metabolism

Reigada et al., 2022

Phenolic Compounds

Sonchus arvensis, Hemerocallis citrina

Improves lifespan & health against nutrient stress

TGF-β signaling pathway; skn-1

An et al., 2023

Cassia fistula Extract

Cassia fistula

Improves survival; reduces ROS & proteotoxicity

DAF-16/FOXO and SKN-1/NRF2 pathways

Thabit et al., 2018

Carbendazim

Fungicide

Inhibits growth & lifespan; damages reproduction & antioxidant systems

Impairs oxidative stress response; toxic

Li et al., 2020

Liuwei Dihuang (LWDH)

Traditional Chinese Medicine

Delays β-amyloid paralysis; reduces ROS

Upregulation of HSPs; antioxidant activity

Sangha et al., 2012

Ethyl Acetate Extract

Gastrodia elata

Protects from oxidative stress & Aβ toxicity; improves lifespan

Insulin/IGF-1 signaling (IIS) pathway

Shi et al., 2023

Acrolein

Toxic Aldehyde

Shortens lifespan; increases ROS; decreases healthspan

Activates DAF-16/FOXO stress response

Jeayeng et al., 2024

Leaf Extract

Anacardium occidentale (Cashew)

Enhances stress survival & lifespan; reduces lipofuscin

DAF-16/FoxO & SKN-1/Nrf-2 pathways; induces sod-3, gst-4

Duangjan et al., 2019

Polysaccharides

Dendrobium officinale

Prolongs lifespan; increases antioxidant enzyme activity

Upregulation of daf-16, skn-1, sir-2.1

Tang et al., 2023

Total Flavonoids

Sea Buckthorn

Increases lifespan (29.4%); enhances stress tolerance; delays paralysis

Radical scavenging; AChE/MAO-A inhibition

Wang et al., 2022

Polysaccharides

Fermented Coix Seed

Increases lifespan (5.9%); enhances antioxidant enzymes

Downregulates daf-2, age-1; upregulates daf-16, sod-3, skn-1

Zhao et al., 2023

Cannabidiol (CBD)

Cannabis sativa

Extends lifespan & survival in AD model

Activates neural glyoxalase pathway; detoxifies methylglyoxal

Frandsen and Narayanasamy, 2022

Extract

Anoectochilus roxburghii

Prolongs lifespan; reduces ROS; increases stress resistance

Activation of daf-16/FoxO pathway

Xu et al., 2024

**Table 2.** Summary of the strengths, limitations, and translational value of zebrafish, fruit fly (*Drosophila melanogaster*), and *Caenorhabditis elegans* in antioxidant research
Model Organism	Key Strengths	Primary Limitations	Translational Value	References
Zebrafish (Danio rerio)	High genetic and physiological similarity to humans (vertebrate model). Transparent embryos enabling real-time, live imaging of developmental and pathological processes. Rapid development and high fecundity. Well-suited for high-throughput genetic and drug screening. High conservation of immune and metabolic pathways.	Limited complexity of some organ systems compared to mammals. High lethality in severe genetic mutations. Differences in drug metabolism and anatomy. Limited genetic background diversity in some strains. Not ideal for long-term chronic studies due to lifespan. Limited studies on complex behaviors and mechanisms.	Highly valuable for drug discovery, toxicology, and safety evaluations. Excellent for modeling human diseases, particularly neurodegenerative, metabolic, and inflammatory conditions. Effective for studying developmental processes and in vivo oxidative stress dynamics.	Ali et al., 2022; Amen et al., 2020; Quelle-Regaldie, et al., 2023; Shreya et al., 2020
Fruit Fly (Drosophila melanogaster)	Extremely short life cycle and rapid generation time. Vast array of well-established, powerful genetic tools for manipulation. Low cost and ease of maintenance. Well-characterized genome. Suitable for high-throughput screening.	Limited physiological and anatomical similarity to humans (invertebrate). Less relevance to complex mammalian systems (e.g., adaptive immunity, complex organ structures). Differences in metabolic pathways. Shorter lifespan can limit long-term studies.	Ideal for initial screening of compounds and for fundamental genetic studies. Provides crucial insights into the genetic basis of aging, oxidative stress mechanisms, and developmental biology. Strong model for gene-environment interactions.	Ajagun-Ogunleye and Ebuehi, 2020; Hamidu et al., 2022; Somegowda et al., 2021
Caenorhabditis elegans (C. elegans)	Simple, well-mapped nervous system and genetics. Transparent body allows for easy observation of cellular processes. Very short lifespan and rapid reproduction are ideal for high-throughput longevity and anti-aging studies. Low maintenance cost. Highly conserved genetic pathways.	Extremely simple anatomy lacks complex organ systems (e.g., no heart, liver, adaptive immune system). Limited relevance to vertebrate physiology and complex human diseases. Short lifespan may not model chronic conditions accurately. Limited behavioral complexity.	Exceptional model for screening antioxidants, anti-aging compounds, and neuroprotective agents. Unparalleled for understanding fundamental cellular mechanisms like apoptosis, stress response, and aging. High value in mechanistic studies of oxidative stress and lifespan extension.	de Araújo, 2025; Ayoub et al., 2024; von Mikecz, 2023; Roxo et al., 2020

Table 2. Summary of the strengths, limitations, and translational value of zebrafish, fruit fly (Drosophila melanogaster), and Caenorhabditis elegans in antioxidant research

Model Organism

Key Strengths

Primary Limitations

Translational Value

References

Zebrafish (Danio rerio)

High genetic and physiological similarity to humans (vertebrate model). Transparent embryos enabling real-time, live imaging of developmental and pathological processes. Rapid development and high fecundity. Well-suited for high-throughput genetic and drug screening. High conservation of immune and metabolic pathways.

Limited complexity of some organ systems compared to mammals. High lethality in severe genetic mutations. Differences in drug metabolism and anatomy. Limited genetic background diversity in some strains. Not ideal for long-term chronic studies due to lifespan. Limited studies on complex behaviors and mechanisms.

Highly valuable for drug discovery, toxicology, and safety evaluations. Excellent for modeling human diseases, particularly neurodegenerative, metabolic, and inflammatory conditions. Effective for studying developmental processes and in vivo oxidative stress dynamics.

Ali et al., 2022; Amen et al., 2020; Quelle-Regaldie, et al., 2023; Shreya et al., 2020

Fruit Fly (Drosophila melanogaster)

Extremely short life cycle and rapid generation time. Vast array of well-established, powerful genetic tools for manipulation. Low cost and ease of maintenance. Well-characterized genome. Suitable for high-throughput screening.

Limited physiological and anatomical similarity to humans (invertebrate). Less relevance to complex mammalian systems (e.g., adaptive immunity, complex organ structures). Differences in metabolic pathways. Shorter lifespan can limit long-term studies.

Ideal for initial screening of compounds and for fundamental genetic studies. Provides crucial insights into the genetic basis of aging, oxidative stress mechanisms, and developmental biology. Strong model for gene-environment interactions.

Ajagun-Ogunleye and Ebuehi, 2020; Hamidu et al., 2022; Somegowda et al., 2021

Caenorhabditis elegans (C. elegans)

Simple, well-mapped nervous system and genetics. Transparent body allows for easy observation of cellular processes. Very short lifespan and rapid reproduction are ideal for high-throughput longevity and anti-aging studies. Low maintenance cost. Highly conserved genetic pathways.

Extremely simple anatomy lacks complex organ systems (e.g., no heart, liver, adaptive immune system). Limited relevance to vertebrate physiology and complex human diseases. Short lifespan may not model chronic conditions accurately. Limited behavioral complexity.

Exceptional model for screening antioxidants, anti-aging compounds, and neuroprotective agents. Unparalleled for understanding fundamental cellular mechanisms like apoptosis, stress response, and aging. High value in mechanistic studies of oxidative stress and lifespan extension.

de Araújo, 2025; Ayoub et al., 2024; von Mikecz, 2023; Roxo et al., 2020