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Article Open Access

Supplementation of aged garlic extract attenuates age-associated memory impairment and cognitive decline: Involvement of molecular pathways in the cortex and hippocampus

  • Authors:
    • Tamanna Mony
    • Marcus Jackson
    • Amitai Zuckerman
    • Wenyan Yu
    • Thao Thi Nguyen
    • Ashley Balderrama
    • Runting Li
    • Grace Y. Sun
    • Jiankun Cui
    • Zezong Gu
  • View Affiliations / Copyright

    Affiliations: University of Missouri School of Medicine, Department of Pathology and Anatomical Sciences, Columbia, MO 65212, USA, Charles W. Gehrke Proteomics Center, University of Missouri, Columbia, MO 65211, USA
    Copyright: © Mony et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 2
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    Published online on: October 27, 2025
       https://doi.org/10.3892/br.2025.2075
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Abstract

The aging of the human population is associated with increases in comorbidities, including functional impairment and cognitive decline. With decreasing numbers of neurons and declining synaptic plasticity, the aging brain exhibits less perfusion, and thus, is more susceptible to attack by reactive oxygen species (ROS). These changes are associated with increased risks of neurodegenerative diseases, which impact the quality of life. Consequently, there is an urgent need to develop nutraceuticals and holistic approaches that may impede the aging process. Aged garlic extract (AGE) is a nutraceutical that has been shown to display neuroprotective properties; in particular, it has the ability to reduce inflammation, apoptosis and ROS. Given these insights, it is reasonable to hypothesize that AGE improves neurological function and cognition by altering neural molecular processes. To test this hypothesis, a battery of behavior tests, including measurements of various cognitive functions, were performed on 82-week-old mice, which were fed a diet supplemented with AGE for 40 weeks. This was followed by label-free global proteomics and machine learning-driven bioinformatics analyses of mouse cortical and hippocampal proteomes for the identification of membrane and cytoskeletal protein targets of the mechanism of action of AGE. Mice in the AGE supplementation group exhibited improvements in learning, memory, exploratory behavior and anxiety (P<0.05), but no obvious changes in other functional domains. Analysis of global proteomes revealed the ability of AGE supplementation to induce expression changes, which were most pronounced in the hippocampus. Further comparative bioinformatics analysis also revealed that AGE supplementation increased synaptogenesis signaling in both brain regions and reduced 14-3-3 signaling related to neuronal apoptosis exclusively in the hippocampus. The predicted upstream global proteome modulators influenced by AGE supplementation included microtubule-associated protein tau, amyloid precursor protein, brain-derived neurotrophic factor, TP53, presenilin 1 and superoxide dismutase 1. Taken together, these findings demonstrated the neuroprotective effects of AGE on cognition and protein expression in aging mice, further suggesting its potential use as a nutraceutical to prevent aging-associated neurological comorbidities.
View Figures

Figure 1

Effects of AGE supplementation on
anxiety and neophobia under adverse light condition. The light-dark
transition test was used to measure (A) number of entries into dark
zone, (B) total time spent in dark zone, (C) number of entries to
light zone, and (D) total time spent in light zone. Data are
presented as the mean ± SEM; Groups: Control, n=24; AGE diet, n=24.
Significantly different from age-matched control group,
*P<0.05 and **P<0.01. AGE, aged garlic
extract.

Figure 2

Effects of AGE supplementation on
neophobia behaviors. The emergence test was used to measure (A)
number of entries into shelter, (B) time spent in shelter, (C)
number of entries into arena, (D) time spent in arena, (E) number
of line crossings in arena, (F) distance travelled in arena and (G)
time immobile in the arena. Data are presented as the mean ± SEM.
Groups: Control, n=24; AGE diet, n=24. Significantly different from
age-matched control group, *P<0.05,
**P<0.01 and ****P<0.0001. AGE, aged
garlic extract; ns, not significant.

Figure 3

AGE effects on NOR. The NOR test was
used to measure (A) Discrimination Index (Scale: -1 to +1), and (B)
Percentage of time investigation with novel object. Data are
represented as the mean ± SEM. Groups: Control, n=24; AGE diet,
n=24. *P<0.05. AGE, aged garlic extract; NOR, novel
object recognition.

Figure 4

AGE effects on cognition and spatial
reference learning. Barnes maze was used to measure latency of
first entering the target zone during habituation and training
days, and reversal training days. Data are presented as mean ± SEM.
Groups: Control, n=24; AGE diet, n=24. On the 2nd reversal training
day (testing day 7), significantly different from age-matched
control group, *P<0.05, analyzed by the one-tailed,
unpaired Student's t-test. AGE, aged garlic extract.

Figure 5

Quantitative proteomics profiling of
cortex and hippocampus in AGE-supplemented mice. (A and B) Volcano
plots showing global proteomic profiles of AGE mice (A) cortex and
(B) hippocampus, respectively; vertical grey lines: -0.34 (decrease
threshold) and +0.34 (increase threshold; horizontal grey lines:
1.3 (significance threshold). Differentially expressed proteins are
colored red (increased expression), blue (decreased expression), or
grey (no change in expression); top 10 hits by Euclidean distance
are labeled. (C) Common and distinct differentially expressed
proteins in the cortex and hippocampus of AGE-supplemented mice are
displayed in the Venn-diagram. Among the identified proteins
exhibiting significant changes, Selenbp1 and Pdyn, were shared
across both brain regions, the cortex and hippocampus, of
AGE-supplemented mice. AGE, aged garlic extract.

Figure 6

IPA analysis of proteomes in
AGE-supplemented mice predicts an altered phenome. (A) Top 10 IPA
canonical pathways in the cortex (red bars) and hippocampus (green)
proteomes of AGE-supplemented mice (P<0.05). (B) Top 10 most
significantly changed upstream regulators in both cortex (left
column) and hippocampus (right column) (P<0.05). Upstream
regulators were ranked by P-value (rather ranking than z-score) in
IPA to assess the statistical enrichment of their downstream
targets. MAPT emerged as a top upstream regulator (P<0.01),
indicating its targets are significantly overrepresented among the
differentially expressed proteins. (C) IPA disease and function
analysis to predict neurological behavior based on differential
protein expression in the cortex and hippocampus (represented by
red lines for increased protein expression and blue lines for
decreased protein expression) indicates changes in behavioral
phenotypes. Differential expression directionality (red lines:
Increasing expression; blue lines: Decreasing expression) in the
cortex and hippocampus predicts altered behavioral phenotypes
including cognition, learning, anxiety, memory and spatial memory
pathways (with red boxes highlighting behaviors driven by increased
expression and blue boxes denoting behaviors influenced by
decreased expression). IPA, Ingenuity Pathway Analysis; AGE, aged
garlic extract; PSEN1, presenilin 1; APP, amyloid precursor
protein.
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Copy and paste a formatted citation
Spandidos Publications style
Mony T, Jackson M, Zuckerman A, Yu W, Nguyen TT, Balderrama A, Li R, Sun GY, Cui J, Gu Z, Gu Z, et al: Supplementation of aged garlic extract attenuates age-associated memory impairment and cognitive decline: Involvement of molecular pathways in the cortex and hippocampus. Biomed Rep 24: 2, 2026.
APA
Mony, T., Jackson, M., Zuckerman, A., Yu, W., Nguyen, T.T., Balderrama, A. ... Gu, Z. (2026). Supplementation of aged garlic extract attenuates age-associated memory impairment and cognitive decline: Involvement of molecular pathways in the cortex and hippocampus. Biomedical Reports, 24, 2. https://doi.org/10.3892/br.2025.2075
MLA
Mony, T., Jackson, M., Zuckerman, A., Yu, W., Nguyen, T. T., Balderrama, A., Li, R., Sun, G. Y., Cui, J., Gu, Z."Supplementation of aged garlic extract attenuates age-associated memory impairment and cognitive decline: Involvement of molecular pathways in the cortex and hippocampus". Biomedical Reports 24.1 (2026): 2.
Chicago
Mony, T., Jackson, M., Zuckerman, A., Yu, W., Nguyen, T. T., Balderrama, A., Li, R., Sun, G. Y., Cui, J., Gu, Z."Supplementation of aged garlic extract attenuates age-associated memory impairment and cognitive decline: Involvement of molecular pathways in the cortex and hippocampus". Biomedical Reports 24, no. 1 (2026): 2. https://doi.org/10.3892/br.2025.2075
Copy and paste a formatted citation
x
Spandidos Publications style
Mony T, Jackson M, Zuckerman A, Yu W, Nguyen TT, Balderrama A, Li R, Sun GY, Cui J, Gu Z, Gu Z, et al: Supplementation of aged garlic extract attenuates age-associated memory impairment and cognitive decline: Involvement of molecular pathways in the cortex and hippocampus. Biomed Rep 24: 2, 2026.
APA
Mony, T., Jackson, M., Zuckerman, A., Yu, W., Nguyen, T.T., Balderrama, A. ... Gu, Z. (2026). Supplementation of aged garlic extract attenuates age-associated memory impairment and cognitive decline: Involvement of molecular pathways in the cortex and hippocampus. Biomedical Reports, 24, 2. https://doi.org/10.3892/br.2025.2075
MLA
Mony, T., Jackson, M., Zuckerman, A., Yu, W., Nguyen, T. T., Balderrama, A., Li, R., Sun, G. Y., Cui, J., Gu, Z."Supplementation of aged garlic extract attenuates age-associated memory impairment and cognitive decline: Involvement of molecular pathways in the cortex and hippocampus". Biomedical Reports 24.1 (2026): 2.
Chicago
Mony, T., Jackson, M., Zuckerman, A., Yu, W., Nguyen, T. T., Balderrama, A., Li, R., Sun, G. Y., Cui, J., Gu, Z."Supplementation of aged garlic extract attenuates age-associated memory impairment and cognitive decline: Involvement of molecular pathways in the cortex and hippocampus". Biomedical Reports 24, no. 1 (2026): 2. https://doi.org/10.3892/br.2025.2075
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