Abstract
Trichostatin A (TSA), a pan-histone deacetylase inhibitor, exerts multiple neuroprotective properties. This study aims to examine whether TSA could enhance autophagy, thereby reduce neuronal apoptosis and ultimately attenuate early brain injury (EBI) following subarachnoid hemorrhage (SAH). SAH was performed through endovascular perforation method, and mortality, neurological score, and brain water content were evaluated at 24 h after surgery. Western blot were used for quantification of acetylated histone H3, LC3-II, LC3-I, Beclin-1, cytochrome c, Bax, and cleaved caspase-3 expression. Immunofluorescence was performed for colocalization of Beclin-1 and neuronal nuclei (NeuN). Apoptotic cell death of neurons was quantified with double staining of terminal deoxynucleotidyl transferase-mediated uridine 5′-triphosphate-biotin nick end-labeling (TUNEL) and NeuN. The autophagy inhibitor 3-methyladenine (3-MA) was used to manipulate the proposed pathway. Our results demonstrated that TSA reduced brain edema and alleviated neurological deficits at 24 h after SAH. TSA significantly increased acetylated histone H3, the LC3-II/LC3-I ratio, and Beclin-1 while decreased Bax and cleaved caspase-3 in the cortex. Beclin-1 and NeuN, TUNEL, and NeuN, respectively, were colocalized in cortical cells. Neuronal apoptosis in the ipsilateral basal cortex was significantly inhibited after TSA treatment. Conversely, 3-MA reversed the beneficial effects of TSA. These results proposed that TSA administration enhanced autophagy, which contributes to alleviation of neuronal apoptosis, improvement of neurological function, and attenuation of EBI following SAH.
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Abbreviations
- SAH:
-
Subarachnoid hemorrhage
- TSA:
-
Trichostatin A
- EBI:
-
Early brain injury
- HDAC:
-
Histone deacetylase
- 3-MA:
-
3-Methyladenine
- TUNEL:
-
Terminal deoxynucleotidyl transferase-mediated uridine 5′-triphosphate-biotin nick end-labeling
- LC3:
-
Light chain 3
- Apaf-1:
-
Apoptotic protease-activating factor 1
- Ac-H3:
-
Acetylated histones H3
- CC3:
-
Cleaved caspase-3
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Acknowledgments
This work was supported by Grant 81371369 and 81371433 from the National Natural Science Foundation of China; Grant 20120101120030 from the Doctoral Program of the Ministry of Education; Grant 2013KYA088 from the Zhejiang Provincial Medical Science and Technology Planning Project; and Grant Y13H090007 and LY13H090002 from the Zhejiang Provincial Natural Science Foundation of China.
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The authors declare that there is no conflicts of interest regarding the publication of this paper.
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Anwen Shao and Zhen Wang contributed equally to this work.
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Shao, A., Wang, Z., Wu, H. et al. Enhancement of Autophagy by Histone Deacetylase Inhibitor Trichostatin A Ameliorates Neuronal Apoptosis After Subarachnoid Hemorrhage in Rats. Mol Neurobiol 53, 18–27 (2016). https://doi.org/10.1007/s12035-014-8986-0
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DOI: https://doi.org/10.1007/s12035-014-8986-0