Silencing of microRNA-494 inhibits the neurotoxic Th1 shift via regulating HDAC2-STAT4 cascade in ischaemic stroke.

Abstract

BACKGROUND AND PURPOSE:

T helper cell 1 (Th1)-skewed neurotoxicity contributes to the poor outcome of stroke in rodents. Here, we have elucidated the mechanism of the Th1/Th2 shift in acute ischaemic stroke (AIS) patients at hyperacute phase and have looked for a miRNA-based therapeutic target.

EXPERIMENTAL APPROACH:

MiR-494 levels in blood from AIS patients and controls were measured by real-time PCR. C57BL/6J mice were subjected to transient middle cerebral artery occlusion, and cortical neurons were subjected to oxygen-glucose deprivation. Luciferase reporter system, chromatin immunoprecipitation sequencing (ChIP-Seq), and ChIP-PCR were used to uncover possible mechanisms.

KEY RESULTS:

In lymphocytes from AIS patients, there was a Th1/Th2 shift and histone deacetylase 2 (HDAC2) was markedly down-regulated. ChIP-seq showed that HDAC2 binding sites were enriched in regulation of Th1 cytokine production, and ChIP-PCR confirmed that HDAC2 binding was changed at the intron of STAT4 and the promoter of T-box transcription factor 21 (T-bet) in lymphocytes from AIS patients. MiR-494 was the most significantly increased miRNA in lymphocytes from AIS patients, and miR-494-3p directly targeted HDAC2. A strong association existed between miR-494 and Th1 cytokines, and neurological deficit as measured by the National Institute of Health Stroke Scale (NIHSS) in AIS patients. In vitro and in vivo experiments showed that antagomir-494 reduced Th1 shift-mediated neuronal and sensorimotor functional damage in the mouse model of ischaemic stroke, via the HDAC2-STAT4 pathway.

CONCLUSION AND IMPLICATIONS:

We demonstrated that miR-494 inhibition prevented Th1-skewed neurotoxicity through regulation of the HDAC2-STAT4 cascade.

• Authors: Haiping Zhao, Guangwen Li, Rongliang Wang, Zhen Tao, Qingfeng Ma, Sijia Zhang, Ziping Han, Feng Yan, Fangfang Li, Ping Liu, Shubei Ma, Xunming Ji, Yumin Luo
• Journal: British journal of pharmacology (Br J Pharmacol) Vol. 177 Issue 1 Pg. 128-144 (01 2020) ISSN: 1476-5381 [Electronic] England
• PMID: 31465536 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2019 The British Pharmacological Society.

Chemical References

• Antagomirs
• MicroRNAs
• Mirn494 microRNA, mouse
• STAT4 Transcription Factor
• Stat4 protein, mouse
• Hdac2 protein, mouse
• Histone Deacetylase 2

Topics

• Aged
• Animals
• Antagomirs (pharmacology)
• Female
• Histone Deacetylase 2 (antagonists & inhibitors, metabolism)
• Humans
• Ischemic Stroke (metabolism, pathology)
• Jurkat Cells
• Male
• Mice
• Mice, Inbred C57BL
• MicroRNAs (antagonists & inhibitors, metabolism)
• Middle Aged
• STAT4 Transcription Factor (antagonists & inhibitors, metabolism)
• Th1 Cells (drug effects, metabolism, pathology)