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Protective effects of Perilla frutescens seed oil on cognitive function, oxidative stress and acetylcholinesterase activity in a D‑galactose‑induced accelerated aging model in rats

  • Authors:
    • Watcharaporn Preedapirom Jeefoo
    • Sitthisak Thongrong
    • Napapan Kangwan
    • Sureena Pohsa
    • Jirayu Kangwan
    • Anongporn Kobroob
    • Rattiya Somka
  • View Affiliations / Copyright

    Affiliations: Division of Physiology, School of Medical Sciences, University of Phayao, Phayao 56000, Thailand, Division of Anatomy, School of Medical Sciences, University of Phayao, Phayao 56000, Thailand, Histology and Pathology Department, Laboratory, Christus St. Vincent Hospital, Santa Fe, NM 87505, USA, Chiang Mai University Demonstration School, Chiang Mai 50200, Thailand, Division of Biochemistry, School of Medical Sciences, University of Phayao, Phayao 56000, Thailand
    Copyright: © Jeefoo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 76
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    Published online on: April 24, 2026
       https://doi.org/10.3892/br.2026.2149
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Abstract

Aging‑related cognitive decline is closely associated with oxidative stress, cholinergic dysfunction and hippocampal vulnerability. Perilla seed oil (PSO), a functional food rich in α‑linolenic acid and antioxidant phytochemicals, may have cognitive benefits; however, its efficacy and underlying mechanisms in experimental models of accelerated aging remain insufficiently understood. The present study aimed to investigate the effects of PSO supplementation on cognitive performance and neurobiological alterations in a D‑galactose (D‑Gal)‑induced accelerated aging model in rats. Wistar rats were injected subcutaneously with D‑Gal (300 mg/kg) daily for 8 weeks and simultaneously treated orally with PSO (100 or 500 mg/kg), fish oil (500 mg/kg) or vehicle. Cognitive function was evaluated using the Morris water maze and novel object recognition tests. Oxidative stress markers, including malondialdehyde (MDA), reduced glutathione (GSH) and superoxide dismutase (SOD), as well as acetylcholinesterase (AChE) activity, were assessed in the hippocampus. Neuronal cell density in the CA1 and CA3 regions was examined using Nissl staining. PSO supplementation significantly improved spatial memory performance and recognition memory, increased hippocampal SOD activity and reduced AChE activity compared with the D‑Gal group. Although MDA and GSH levels did not differ significantly, both exhibited a tendency toward normalization. In addition, neuronal density in the CA3 region was significantly reduced in the D‑Gal group compared with the control group, whereas no significant differences were observed in the CA1 region. These findings suggest that PSO attenuates D‑Gal‑induced cognitive impairment, which may be partially associated with enhanced antioxidant enzyme activity and modulation of cholinergic function, rather than with restoration of neuronal density. PSO may therefore represent a potential nutritional intervention for supporting cognitive function during aging‑related neurobiological changes.
View Figures

Figure 1

Experimental design of the animal
study. Male Wistar rats were randomly assigned to five groups:
Control sham, D-Gal, D-Gal plus FO and D-Gal + PSO at doses of 100
or 500 mg/kg. Treatments were administered for 8 weeks. Cognitive
function was evaluated using the MWM (days 50-54), followed by a
probe trial (day 55) and the NOR test (day 56) prior to sacrifice.
CO, corn oil; D-Gal, D-galactose; FO, fish oil; MWM, Morris water
maze; NOR, novel object recognition; PSO, Perilla seed oil.

Figure 2

Effects of PSO on spatial memory in
rats. (A) Escape latency during the Morris water maze training
trials. (B) Time spent in the target quadrant during the probe
trial. Data are presented as the mean ± SEM. **P<0.01
and ***P<0.001 compared with the D-Gal group. D-Gal,
D-galactose; FO500, fish oil 500 mg/kg; PSO100, Perilla seed oil
100 mg/kg; PSO500, Perilla seed oil 500 mg/kg.

Figure 3

Effect of PSO on recognition memory
in rats. (A) Exploration time of the two identical objects (A1 and
A2) during the training phase of the novel object recognition test.
(B) Exploration time of the familiar object (A) and the novel
object (B) during the test phase. (C) DR calculated as (TB-TA)/(TB
+ TA). Data are presented as the mean ± SEM. In panel B,
*P<0.05 indicates a significant within-group
difference between the familiar and novel objects (paired Student's
t-test). In panel C, *P<0.05, **P<0.01
and ****P<0.0001 compared with the D-Gal group
(one-way ANOVA followed by Dunnett's post hoc test). D-Gal,
D-galactose; DR, discrimination ratio; FO500, fish oil 500 mg/kg;
PSO100, Perilla seed oil 100 mg/kg; PSO500, Perilla seed oil 500
mg/kg.

Figure 4

Effects of PSO on oxidative stress
markers and AChE activity in the hippocampus. (A) MDA levels. (B)
GSH levels. (C) SOD activity. (D) AChE activity in the hippocampus.
Data are presented as the mean ± SEM. *P<0.05,
**P<0.01 and ***P<0.001 compared with
the D-Gal group. AChE, acetylcholinesterase; D-Gal, D-galactose;
FO500, fish oil 500 mg/kg; PSO100, Perilla seed oil 100 mg/kg;
PSO500, Perilla seed oil 500 mg/kg; GSH, reduced glutathione; MDA,
malondialdehyde; SOD, superoxide dismutase.

Figure 5

Effects of PSO on hippocampal
neuronal cell density in CA1 and CA3 regions. (A) Representative
Nissl-stained images of the CA1 and CA3 pyramidal cell layers. (B)
Quantification of neuronal cell density in the CA1 region
(cells/mm²). (C) Quantification of neuronal cell density in the CA3
region (cells/mm²). Data are presented as the mean ± SEM.
***P<0.001 compared with the D-Gal group. Scale bar,
50 µm. D-Gal, D-galactose; FO500, fish oil 500 mg/kg; PSO100,
Perilla seed oil 100 mg/kg; PSO500, Perilla seed oil 500 mg/kg.
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Jeefoo WP, Thongrong S, Kangwan N, Pohsa S, Kangwan J, Kobroob A and Somka R: Protective effects of <em>Perilla frutescens</em> seed oil on cognitive function, oxidative stress and acetylcholinesterase activity in a D‑galactose‑induced accelerated aging model in rats. Biomed Rep 24: 76, 2026.
APA
Jeefoo, W.P., Thongrong, S., Kangwan, N., Pohsa, S., Kangwan, J., Kobroob, A., & Somka, R. (2026). Protective effects of <em>Perilla frutescens</em> seed oil on cognitive function, oxidative stress and acetylcholinesterase activity in a D‑galactose‑induced accelerated aging model in rats. Biomedical Reports, 24, 76. https://doi.org/10.3892/br.2026.2149
MLA
Jeefoo, W. P., Thongrong, S., Kangwan, N., Pohsa, S., Kangwan, J., Kobroob, A., Somka, R."Protective effects of <em>Perilla frutescens</em> seed oil on cognitive function, oxidative stress and acetylcholinesterase activity in a D‑galactose‑induced accelerated aging model in rats". Biomedical Reports 24.6 (2026): 76.
Chicago
Jeefoo, W. P., Thongrong, S., Kangwan, N., Pohsa, S., Kangwan, J., Kobroob, A., Somka, R."Protective effects of <em>Perilla frutescens</em> seed oil on cognitive function, oxidative stress and acetylcholinesterase activity in a D‑galactose‑induced accelerated aging model in rats". Biomedical Reports 24, no. 6 (2026): 76. https://doi.org/10.3892/br.2026.2149
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Spandidos Publications style
Jeefoo WP, Thongrong S, Kangwan N, Pohsa S, Kangwan J, Kobroob A and Somka R: Protective effects of <em>Perilla frutescens</em> seed oil on cognitive function, oxidative stress and acetylcholinesterase activity in a D‑galactose‑induced accelerated aging model in rats. Biomed Rep 24: 76, 2026.
APA
Jeefoo, W.P., Thongrong, S., Kangwan, N., Pohsa, S., Kangwan, J., Kobroob, A., & Somka, R. (2026). Protective effects of <em>Perilla frutescens</em> seed oil on cognitive function, oxidative stress and acetylcholinesterase activity in a D‑galactose‑induced accelerated aging model in rats. Biomedical Reports, 24, 76. https://doi.org/10.3892/br.2026.2149
MLA
Jeefoo, W. P., Thongrong, S., Kangwan, N., Pohsa, S., Kangwan, J., Kobroob, A., Somka, R."Protective effects of <em>Perilla frutescens</em> seed oil on cognitive function, oxidative stress and acetylcholinesterase activity in a D‑galactose‑induced accelerated aging model in rats". Biomedical Reports 24.6 (2026): 76.
Chicago
Jeefoo, W. P., Thongrong, S., Kangwan, N., Pohsa, S., Kangwan, J., Kobroob, A., Somka, R."Protective effects of <em>Perilla frutescens</em> seed oil on cognitive function, oxidative stress and acetylcholinesterase activity in a D‑galactose‑induced accelerated aging model in rats". Biomedical Reports 24, no. 6 (2026): 76. https://doi.org/10.3892/br.2026.2149
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