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Neuroprotective effect of Continentalic acid by targeting NF-κB/Bcl-2 and oxidative stress signaling in traumatic brain injury in male BALB/c mice

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Abstract

Background

Traumatic brain injury (TBI) is characterized as a heterogeneous neurological disorder, which causes a variety of behavioral and functional impairments. Continentalic acid (CA), a diterpenoid acid obtained from Aralia continentalis, possesses anti-inflammatory, anti-apoptotic, and antioxidant properties.

Methods

In the current study we investigated the neuroprotective properties of CA in an experimental TBI model. An in vivo, weight-drop model of TBI was developed in male BALB/c mice. Following traumatic insult, CA (1, 10, and 50 mg/kg) was administered intraperitoneally for up to 14 days. To assess the protective effects of CA, we performed behavioral tests, spectroscopy, histopathology, and biochemical analyses.

Results

The results revealed that CA significantly attenuated trauma-evoked neurological functions and improved mechanical as well as periorbital allodynia. CA also restored the structural and molecular composition of traumatized brain tissues. Similarly, CA reversed the histological changes in the cortex region of the traumatized brain sections and improved the neuronal count. CA administration also decreased the level of traumatic microbleeds in the brain. The expression levels of GFAP and Iba1 were significantly reduced by CA. Treatment with CA significantly reduced the expression of TLR-4, NF-κB, and increased the expression of IĸB-α regulatory protein. CA also downregulated the expression of cleaved caspase-3 protein and upregulated the expression of anti-apoptotic Bcl-2 protein and alleviated the DNA damage. CA dramatically improved the levels of GST, GSH, catalase, and SOD while concomitantly reducing NO and LPO levels. The levels of IL-1β and TNF-α were also reduced in the treatment group.

Conclusions

The results demonstrated that CA (50 mg/kg) exhibited neuroprotective activity in TBI by attenuating inflammation, neuronal apoptosis, and oxidative stress.

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Data availability

All the RAW data of the respective experiments was provided as a supplementary file.

Abbreviations

CDNB:

1-chloro-24-dinitrobenzene

DTNB:

5–5’dithio-bis-2-nitro benzoic acid

ABC:

Avidin-biotin-peroxidase complex

Bcl-2:

B-cell lymphoma 2

BBB:

Blood Brain Barrier

JNK:

c-Jun N-terminal kinase

DAB:

Diaminobenzidine

DMSO:

Dimethylsulfoxide

FTIR:

Fourier Transform Infrared Spectroscopy

GFAP:

Glial fibrillary acidic protein

GST:

Glutathione-S-transferase

H&E:

Hematoxylin and eosin

HO-1:

Heme Oxygenase-1

H2O2:

Hydrogen peroxide

IL-1β:

Interleukin-1beta

Iba1:

Ionized calcium-binding adapter molecule 1

LPO:

Lipid peroxidase

MAPK:

Mitogen-Activated Protein Kinase

NO:

Nitric oxide

Nrf2:

Nuclear factor erythroid 2-related factor 2

NF-kB:

Nuclear factor-kappa-B

PWT:

Paw Withdrawal Threshold

PT:

Periorbital Threshold

PGE2:

Prostaglandin E2

qRT-PCR:

Quantitative real-time polymerase chain reaction

GSH:

Reduced glutathione

SOD:

Superoxide dismutase

TBA:

Thiobarbituric acid

TLR-4:

Toll-like receptor 4

TBI:

Traumatic brain injury

TNF-α:

Tumor necrosis factor-alpha

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Funding

The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.

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Authors and Affiliations

  1. Pharmacological Sciences Research Lab, Department of Pharmacy, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, 45320, Pakistan

    Sana Zafar, Maryam Jamil, Muhammad Ibrar Khan & Salman Khan

  2. Department of Pharmacy, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, 45320, Pakistan

    Sana Zafar, Maryam Jamil, Muhammad Ibrar Khan, Kifayat Ullah Shah & Salman Khan

  3. Department of Pharmacology, Faculty of Pharmacy, Capital University of Science & Technology, Islamabad Expressway, Islamabad, 747424, Pakistan

    Muhammad Ibrar Khan

  4. Department of Pharmacy, City University of Science and Information Technology, Peshawar, KP, Pakistan

    Ashraf Ullah Khan

  5. College of Pharmacy, Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul, 03760, South Korea

    Eun Kyoung Seo

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  1. Sana Zafar
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Contributions

S Z, M J, M I K, A U K, and S K contributed to the study conception and design. Material preparation, data collection, and analysis were performed by S Z, M J, M I K, K U S, E K S. Project administration, supervision were performed by S K. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Salman Khan.

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Competing interests

The authors declare no competing interests.

Ethical approval

The experiment was performed according to the guidelines issued by the National Institute of Health for the Care and Use of Laboratory Animals, along with approval from the bio-ethical committee of Quaid-i-Azam University assigned with approval number BEC-FBS-QAU2023-460.

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Zafar, S., Jamil, M., Khan, M.I. et al. Neuroprotective effect of Continentalic acid by targeting NF-κB/Bcl-2 and oxidative stress signaling in traumatic brain injury in male BALB/c mice. Mol Biol Rep 53, 135 (2026). https://doi.org/10.1007/s11033-025-11294-5

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