Hesperetin Confers Neuroprotection by Regulating Nrf2/TLR4/NF-κB Signaling in an Aβ Mouse Model

  • Muhammad Ikram
  • Tahir Muhammad
  • Shafiq Ur Rehman
  • Amjad Khan
  • Min Gi Jo
  • Tahir Ali
  • Myeong Ok KimEmail author


Hesperetin is a bioactive flavonoid in the body, produced from hesperidin. No comprehensive studies have shown its protective effects in neurodegenerative disorders. Here, we hypothesized that hesperetin may protect the mice brain against Aβ-induced neurodegeneration. Twenty-four hours after intracerebroventricular injection of Aβ1-42, the treated group was injected hesperetin. For in vitro experiments, HT22 and BV-2 cells were used. Immunoblot, immunofluorescence, and behavioral analyses were used to evaluate the different parameters. Our results indicated that hesperetin significantly attenuated oxidative stress, as assessed by the expression of Nrf2/HO-1 and LPO and ROS assays, in the hippocampus, cortex, and in vitro HT22 cells. Similarly, activated glial cells were regulated by hesperetin, as assessed by the expression of GFAP and Iba-1. Moreover, the expression of TLR4, p-NF-κB, and downstream targets was analyzed; the results showed that hesperetin reinstated the expression of these markers. The effects of hesperetin were further confirmed by using specific TLR4 and p-NF-κB inhibitors in BV-2 cells. Next, we evaluated Aβ pathology in the cortex, hippocampus, and HT22 cells, showing that hesperetin significantly reduced the Aβ pathology. Furthermore, the antiapoptotic effects of hesperetin were assessed, which showed strong antiapoptotic effects. Overall, the neuroprotective effect of hesperetin was found to be a multipotent effect, involving the inhibition of oxidative stress, neuroinflammation, apoptotic cell death, and cognitive consolidation. Given antioxidant, anti-inflammatory, and antiapoptotic potentials against Aβ-induced neurodegeneration and memory impairment, hesperetin may be a promising therapeutic agent for Alzheimer’s disease–like neurological disorders.


Amyloid beta Neuroinflammation Neurodegeneration Hesperetin Neuroprotection 



nuclear factor erythroid 2-related factor 2


heme oxygenase 1


Toll-like receptor 4


tissue necrosis factor-α


Alzheimer’s disease


central nervous system

amyloid beta


phosphate buffer saline


sodium dodecyl sulfate-polyacrylamide gel electrophoresis


fluorescein isothiocyanate


tetramethylrhodamine isothiocyanate




Author Contributions

MI: Concept, design, data collection, analysis, interpretation, and manuscript writing.

TM: Mice grouping, treatment, and performed in vitro experiments.

SUR: Mice grouping, manuscript evaluation, and interpretation.

AK: Mice treatment and behavioral studies.

TA: Data collection and analysis.

MGJ: Behavioral studies, animal handling, and treatment.

MOK: The corresponding author, reviewed and approved the manuscript, and holds all the responsibilities related to this manuscript. All the authors reviewed the revised manuscript.

Funding Information

This research work was supported by the Brain Research Program through the National Research Foundation of Korea funded by the Ministry of Science, ICT & Future Planning (2016M3C7A1904391).

Compliance with Ethical Standards

The animal maintenance, treatments, behavioral studies, and surgical procedures were carried out in accordance with the animal ethics committee (IACUC) guidelines issued by the Division of Applied Life Sciences, Department of Biology at Gyeongsang National University, South Korea. The experimental methods were carried out in accordance with the approved 22 guidelines (Approval ID: 125) and all experimental protocols were approved by the animal ethics committee (IACUC) of the Division of Applied Life Sciences, Department of Biology at Gyeongsang National University, South Korea.

Conflict of Interest

The authors declare that they have no conflict of interest.


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Authors and Affiliations

  1. 1.Division of Life Science and Applied Life Science (BK21 plus), College of Natural SciencesGyeongsang National UniversityJinjuRepublic of Korea

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