Abstract
In recent years, orexin (ORX) and melanin-concentrating hormone (MCH) have been demonstrated to exert neuroprotective roles in cerebral ischemia. Hence, this study investigated the regulatory function of ORX and MCH in neurological function following ischemic stroke and explored the molecular mechanism underlying these functions. A rat model of ischemic stroke was developed by middle cerebral artery occlusion (MCAO), and Longa scoring was employed to evaluate the degree of neurological function deficit. The expression patterns of ORX and MCH were examined by real-time polymerase chain reaction in the brain tissues of rats with ischemic stroke induced by middle cerebral artery occlusion (MCAO). Moreover, electroencephalography (EEG) analysis and high-performance liquid chromatography (HPLC) were respectively performed to detect rapid-eye movement (REM) sleep, the glutamate (Glu) uptake, and the expression of γ-aminobutyric acid B receptor (GABAB). Immunoblotting was performed to test the levels of autophagic markers LC3, BECLIN-1, and p62. Immunohistochemistry (IHC) staining and TUNEL assays were respectively used to assess the autophagy and neuronal apoptosis. Results demonstrated that ORX and MCH were lowly expressed in brain of rats with ischemic stroke. ORX or MCH overexpression decreased neuronal apoptosis and autophagy, and improved the sleep architecture of post-stroke rats, while rescuing Glu uptake and GABA expression. ORX or MCH upregulation exerted protective effects on neurological function. Taken together, ORX and/or MCH protect against ischemic stroke in a rat model, highlighting their value as targets for the clinical treatment of ischemic stroke.
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Abbreviations
- ORX:
-
Orexin
- MCH:
-
Melanin concentrating hormone
- MCAO:
-
Middle cerebral artery occlusion
- EEG:
-
Electroencephalography
- HPLC:
-
High-performance liquid chromatography
- REM:
-
Rapid-eye movement
- Glu:
-
Glutamate
- GABAB :
-
γ-Aminobutyric acid B receptor
- IHC:
-
Immunohistochemistry
- p62:
-
Protein sequestosome 1/p62
- oe:
-
Overexpression
- Tturn :
-
Time taken by the rats to orient themselves 180° head-downwards
- Ttotal :
-
Time taken to place all four paws on the ground reached the ground
- TUNEL:
-
Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling
- DAPI:
-
6-Diamidino-2-phenylindole
- TTC:
-
2, 3, 5-Triphenyltetrazolium chloride
- OPA:
-
O-phthalaldehyde
- RT-qPCR:
-
Reverse transcription quantitative polymerase chain reaction
- SDS:
-
Sodium dodecyl sulfate
- OX1R:
-
Orexin-1-receptor
- OX2R:
-
Orexin-2-receptor
- TBST:
-
Tris-buffered saline Tween
- ANOVA:
-
Analysis of variance
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Funding
This study was supported by the Fundamental Research Funds for the Central Universities (No. 012019121); Xi’an Social Development Guidance Plan Medical Research Project (No. 2017115SF/YX009); and Natural Science Foundation of Shaanxi Province (2021JM-567; 2021JQ-923).
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Qing Shu and Gang Wu designed the study. Xi’an Zhang and Shijun Li collated the data, carried out data analyses, and produced the initial draft of the manuscript. Dan Zhou, Jie Bai, and Hanxiang Wang contributed to drafting the manuscript. All authors have read and approved the final submitted manuscript.
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Wu, G., Zhang, X., Li, S. et al. Overexpression of ORX or MCH Protects Neurological Function Against Ischemic Stroke. Neurotox Res 40, 44–55 (2022). https://doi.org/10.1007/s12640-021-00457-4
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DOI: https://doi.org/10.1007/s12640-021-00457-4