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Clinacanthus nutans Mitigates Neuronal Apoptosis and Ischemic Brain Damage Through Augmenting the C/EBPβ-Driven PPAR-γ Transcription

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Abstract

Clinacanthus nutans Lindau (C. nutans) is a traditional herbal medicine widely used in Asian countries for treating a number of remedies including snake and insect bites, skin rashes, viral infections, and cancer. However, the underlying molecular mechanisms for its action and whether C. nutans can offer protection on stroke damage in brain remain largely unknown. In the present study, we demonstrated protective effects of C. nutans extract to ameliorate neuronal apoptotic death in the oxygen-glucose deprivation model and to reduce infarction and mitigate functional deficits in the middle cerebral artery occlusion model, either administered before or after hypoxic/ischemic insult. Using pharmacological antagonist and siRNA knockdown approaches, we demonstrated ability for C. nutans extract to protect neurons and ameliorate ischemic injury through promoting the anti-apoptotic activity of peroxisome proliferator-activated receptor-gamma (PPAR-γ), a stress-induced transcription factor. Reporter and chromatin immunoprecipitation promoter analysis further revealed C. nutans extract to selectively increase CCAAT/enhancer binding protein (C/EBP)β binding to specific C/EBP binding site (-332~-325) on the PPAR-γ promoter to augment its transcription. In summary, we report a novel transcriptional activation involving C/EBPβ upregulation of PPAR-γ expression to suppress ischemic neuronal apoptosis and brain infarct. Recognition of C. nutans to enhance the C/EBPβ → PPAR-γ neuroprotective signaling pathway paves a new way for future drug development for prevention and treatment of ischemic stroke and other neurodegenerative diseases.

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Abbreviations

ACO:

Acyl-CoA oxidase

C. nutans :

Clinacanthus nutans Lindau

CN:

80% ethanol leaf extract of C. nutans

CNS:

Central nervous system

C/EBP:

CCAAT/enhancer-binding protein

ChIP:

Chromatin immunoprecipitation

DMSO:

Dimethyl sulfoxide

β-Gal:

β-Galactosidase

icv:

Intracerebroventricular

i.p.:

Intraperitoneal

MCA:

Middle cerebral artery

MRI:

Magnetic resonance imaging

OGD:

Oxygen-glucose deprivation

PNs:

Primary cortical neurons

PPAR-γ:

Peroxisome proliferator-activated receptor-gamma

PPRE:

PPAR response element

tPA:

Tissue-type plasminogen activator

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Acknowledgements

We thank the Taiwan Mouse Clinic (TMC; http://tmc.sinica.edu.tw/index.html; MRI: Yu-Ying Tung) for technical support. All authors have read and agreed to the manuscript as written.

Funding Information

This work was supported by grants from the Minister of Science and Technology and Academia Sinica of Taiwan to T.N. Lin.

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Authors and Affiliations

  1. Institute of Biomedical Sciences, Academia Sinica, Taipei, 11529, Taiwan, Republic of China

    Jui-Sheng Wu, Mei-Han Kao, Hsin-Da Tsai, Wai-Mui Cheung, Jin-Jer Chen & Teng-Nan Lin

  2. Department of Anatomy, National University of Singapore, Singapore, Singapore

    Wei-Yi Ong

  3. Department of Biochemistry, University of Missouri, Columbia, MO, USA

    Grace Y. Sun

  4. Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Republic of China

    Teng-Nan Lin

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  1. Jui-Sheng Wu
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Wu, JS., Kao, MH., Tsai, HD. et al. Clinacanthus nutans Mitigates Neuronal Apoptosis and Ischemic Brain Damage Through Augmenting the C/EBPβ-Driven PPAR-γ Transcription. Mol Neurobiol 55, 5425–5438 (2018). https://doi.org/10.1007/s12035-017-0776-z

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