Pivotal role of cerebral interleukin-23 during immunologic injury in delayed cerebral ischemia in mice

Y Zheng; D Zhong; H Chen; S Ma; Y Sun; M Wang; Q Liu; G Li

doi:10.1016/j.neuroscience.2015.01.041

Pivotal role of cerebral interleukin-23 during immunologic injury in delayed cerebral ischemia in mice

Neuroscience. 2015 Apr 2:290:321-31. doi: 10.1016/j.neuroscience.2015.01.041. Epub 2015 Jan 28.

Authors

Y Zheng¹, D Zhong¹, H Chen¹, S Ma¹, Y Sun¹, M Wang¹, Q Liu¹, G Li²

Affiliations

¹ Department of Neurology, The First Affiliated Hospital, Harbin Medical University, 23 You Zheng Street, Harbin 150001, Heilong Jiang Province, PR China.
² Department of Neurology, The First Affiliated Hospital, Harbin Medical University, 23 You Zheng Street, Harbin 150001, Heilong Jiang Province, PR China. Electronic address: hydlgz1962@163.com.

PMID: 25637493
DOI: 10.1016/j.neuroscience.2015.01.041

Abstract

Background: Interleukin-23 (IL-23) is required for T helper 17 (Th17) cell responses and IL-17 production in ischemic stroke. We previously showed that the IL-23/IL-17 axis aggravates immune injury after cerebral infarction in mice. However, IL-23 might activate other cytokines and transcription factor forkhead box P3 (Foxp3) production in cerebral ischemia. We aimed to determine whether IL-23p19 knockdown prevents cerebral ischemic injury by reducing ischemic-induced inflammation.

Methods: Ischemic stroke models were established by permanent middle cerebral arterial occlusion (pMCAO) in male C57BL/6 mice. In vivo gene knockdown was achieved by intravenous delivery of lentiviral vectors (LVs) encoding IL-23p19 short hairpin RNA (LV-IL-23p19 shRNA). Enzyme-linked immunoassay (ELISA) and quantitative real-time polymerase chain reaction (qRT-PCR) confirmed inhibitory efficiency. Behavioral deficits were evaluated by adhesive-removal somatic-sensory test. Brain infarct volume was measured at day 5 after pMCAO by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Expression of IL-17, IL-4, interferon (IFN)-γ and Foxp3 in ischemic brain tissues were detected by qRT-PCR and Western blotting, respectively. Additionally, immunohistochemical staining located cytokines in ischemic brain tissues.

Results: RNA interference knockdown of IL-23p19 resulted in improved neurological function and reduced infarct volume. IL-23p19 knockdown suppressed IL-17 gene and protein expression. Moreover, IL-23p19 deficiency enhanced IFN-γ and Foxp3 expressions in delayed cerebral ischemic mice, and did not impact IL-4 expression. Immunohistochemical staining showed that IL-17, IL-4, IFN-γ and Foxp3-positive cells were located around ischemic lesions of the ipsilateral hemisphere.

Conclusions: IL-23p19 knockdown prevents delayed cerebral ischemic injury by dampening the ischemia-induced inflammation, and is a promising approach for clinically managing ischemic stroke.

Keywords: Foxp3; IFN-γ; IL-17; IL-23p19 knockdown; cerebral ischemia; pMCAO.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Brain / metabolism*
Brain / pathology
Brain Ischemia / metabolism*
Brain Ischemia / pathology
Disease Models, Animal
Forkhead Transcription Factors / metabolism
Gene Knockdown Techniques
Infarction, Middle Cerebral Artery
Interferon-gamma / metabolism
Interleukin-17 / metabolism
Interleukin-23 Subunit p19 / genetics
Interleukin-23 Subunit p19 / metabolism*
Male
Mice, Inbred C57BL
RNA, Messenger / metabolism
RNA, Small Interfering
Random Allocation
Stroke / metabolism
Stroke / pathology
Time Factors

Substances

Forkhead Transcription Factors
Foxp3 protein, mouse
Interleukin-17
Interleukin-23 Subunit p19
RNA, Messenger
RNA, Small Interfering
Interferon-gamma