Dexmedetomidine preconditioning ameliorates lung injury induced by pulmonary ischemia/reperfusion by upregulating promoter histone H3K4me3 modification of KGF-2

doi:10.1016/j.yexcr.2021.112762

Experimental Cell Research

Volume 406, Issue 2, 15 September 2021, 112762

https://doi.org/10.1016/j.yexcr.2021.112762 Get rights and content

Highlights

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DexP alleviates I/R-induced lung injury and endothelial barrier dysfunction in mice.
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DexP reduces the inflammatory response and increases KGF-2 expression in I/R.
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DexP regulates the H3K4me3 modification of KGF-2 promoter histone.
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DexP promotes KGF-2 expression by downregulating the expression of JMJD3.
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DexP has the potential to be used as a means of treating I/R-induced lung injury.

Abstract

Keratinocyte growth factor (KGF)-2 has been highlighted to play a significant role in maintaining the endothelial barrier integrity in lung injury induced by ischemia-reperfusion (I/R). However, the underlying mechanism remains largely unknown. The aims of this study were to determine whether dexmedetomidine preconditioning (DexP) modulates pulmonary I/R-induced lung injury through the alteration in KGF-2 expression. In our I/R-modeled mice, DexP significantly inhibited pathological injury, inflammatory response, and inflammatory cell infiltration, while promoted endothelial barrier integrity and KGF-2 promoter activity in lung tissues. Bioinformatics prediction and ChIP-seq revealed that I/R significantly diminished the level of H3K4me3 modification in the KGF-2 promoter, which was significantly reversed by DexP. Moreover, DexP inhibited the expression of histone demethylase JMJD3, which in turn promoted the expression of KGF-2. In addition, overexpression of JMJD3 weakened the protective effect of DexP on lung injury in mice with I/R. Collectively, the present results demonstrated that DexP ameliorates endothelial barrier dysfunction via the JMJD3/KGF-2 axis.

Graphical abstract

Introduction

Lung transplantation, acute lung injury, and acute respiratory distress syndrome can cause ischemia/reperfusion (I/R)-related lung injury [1]. After reperfusion, the endothelial layer of the pulmonary vasculature yields reactive oxygen species and other harming agents, which contributes to increased microvascular permeability, the major step in modulating I/R injury [2]. There are no therapeutic agents clinically employed to prevent I/R-induced lung injury, and treatment options are limited to supportive care [3]. Consequently, it is of great importance to illuminate the underlying mechanisms of endothelial barrier dysfunction in lung injury induced by I/R.

Dexmedetomidine (Dex), a highly specific α2-adrenergic agonist, possesses anesthetic, analgesia and sympatholytic properties and could mediate gene expression, inflammatory processes and apoptotic cell death [4]. Dex has been suggested to protect against hepatic and cardiac I/R injury in mice and rats, respectively [5,6]. More importantly, Dex has been indicated to attenuate the production of proinflammatory cytokines to protect the lung from I/R injury [7]. However, the relationship between Dex and endothelial barrier dysfunction in lung I/R injury remains to be established. Previously, pretreatment with keratinocyte growth factor (KGF)-2 has been demonstrated to appreciably prevent I/R-induced lung edema, inflammatory cell infiltration, as well as endothelial cell apoptosis and barrier dysfunction [8]. Therefore, we wondered whether the protective effects of Dex in the lung was in a KGF-2-dependent manner. KGF-2, also termed as fibroblast growth factor-10 (FGF-10), has been implicated in lung development and in preventing lung injury from multiple stresses in rats [9]. Interestingly, the participation of epigenetic modifiers in the events of lung inflammation and protection of microvascular permeability has been increasingly underscored [10]. Moreover, a global suppression of histone methylation, specifically H3K4me3, has been identified and validated in neonatal hyperoxia exposure [11]. Among them, jumonji domain-containing protein 3 demethylase (JMJD3), namely lysine-specific demethylase 6B (KDM6B), is favorably recruited to transcription start sites featured by high modification levels of H3K4me3, which is typically associated with gene activation [[12], [13], [14]]. Interestingly, Dex has been previously exhibited to alleviate astrocyte apoptosis via the inhibition of JMJD3 expression [15]. Thus, we hypothesized that Dex preconditioning (DexP) alleviated I/R-induced lung injury by activating the KGF-2 promoter activity via JMJD3. In this study, we established a mouse model of lung injury induced by I/R and a human pulmonary microvascular endothelial cell (HPMEC) model exposed to oxygen-glucose deprivation/reoxygenation (OGD/R) with an aim to investigate the effects of DexP on I/R-induced lung injury and its potential mechanisms.

Section snippets

Antibodies and reagents

Antibodies targeting VCAM-1 (#GTX64226, GeneTex, Inc., Alton Pkwy Irvine, CA, USA), MMP9 (#MA5-15886, Invitrogen Inc., Carlsbad, CA, USA), Cleaved Caspase-3 (#PA5-114687, Invitrogen), Bcl-2 (#GTX100064, GeneTex), SP-C (#PA5-71680, Invitrogen), VEGF (#MA5-13182, Invitrogen), H3K4me2 (ab32356, Abcam, Cambridge, UK), H3K4me3 (ab8580, Abcam), JMJD3 (#GTX124222, GeneTex), CD31 (ab28364, Abcam), and vWF (ab6994) were used. ELISA kits for TNF-ɑ (#MTA00B), IL-1β (#MLB00C), and IL-6 (#ML600B) were

DexP alleviates I/R-induced lung injury and endothelial barrier dysfunction in mice

First, we developed a mouse model with lung injury by I/R and examined the pathological changes in mouse lung tissues by HE staining. In the lung tissues of surgically treated mice, the gaps between cells became blurred and there was obvious inflammatory cell infiltration (Fig. 1A). Moreover, immunohistochemical detection of apoptosis-related proteins revealed a significant elevation in the staining intensity of Cleaved caspase-3 and a significant decrease in the staining intensity of Bcl-2 in

Discussion

There is emerging evidence suggesting that Dex exerts protective effects against I/R-induced injuries on various organs, including the spinal cord, brain, retina, as well as lung through mediating inflammatory responses and/or cell apoptosis/autophagy [[19], [20], [21], [22]]. OGD/R-evoked inflammation and the relevant pulmonary endothelial injury are the core processes in I/R-related lung injury and can result in destruction of the pulmonary barrier, which promotes its permeability [23].

Conclusion

Our data showed that DexP treatment provided a protective effect on lung injury induced by I/R, with the protective effects attributed principally to the downregulation of JMJD3 and the subsequent activation of KGF-2 expression (Fig. 8). The current findings provide a basis for the development of new strategies for the treatment of lung injury. Nevertheless, further studies are needed to confirm how DexP causes the reduced expression of JMJD3. A potential limitation of our study is that we only

Credit author statement

Huisuo Hong and Qingqing Huang: Conceptualization, Methodology, Experimental studies, Software; Yaoyao Cai and Zhousheng Jin: Data curation, Statistical analysis; Tingting Lin: Visualization, Experimental studies; Fangfang Xia: Manuscript preparation, Reviewing and Editing, Validation; All authors read and approved the final manuscript.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

This work was supported by the Zhejiang Provincial Natural Science Foundation of China under (Grant No. LWQ20H310001).

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Chronic obstructive pulmonary disease (COPD), a heterogeneity of chronic respiratory disease pattern, presents considerable prevalence and mortality. We aimed to explore the role and mechanisms of Jumonji domain-containing protein-3 (JMJD3) in COPD. The viability and JMJD3 expression in human bronchial epithelial cell line BEAS-2B were respectively assayed by CCK-8 assay and Western blot following stepwise exposure to increasing concentrations of cigarette smoke extract (CSE). After JMJD3 was silenced and acyl-CoA synthetase long-chain family member 4 (ACSL4) was overexpressed in CSE-treated BEAS-2B cells, cell viability, cytotoxicity, oxidative stress and total iron level were estimated using kits. ELISA estimated inflammatory levels. DCFH-DA probe and BODIPY 581/591 C11 probe were exposed to assess ROS production and lipid peroxidation. Western blot tested the expressions of ferroptosis-associated proteins. Besides, H3K27me3 and ACSL4 expressions were tested by Western blot and immunofluorescence staining. In CSE-induced BEAS-2B cells, JMJD3 expression was increased and deletion of JMJD3 improved cell viability, reduced LDH release, mitigated inflammation, oxidative stress and inhibited ferroptosis. Moreover, JMJD3 interference raised H3K27me3 expression whereas lessened ACSL4 expression in CSE-treated BEAS-2B cells. CSE exposure reduced the abundance of ACSL4 in H3K27me3 antibody. Further ACSL4 elevation reversed the impacts of JMJD3 silencing on the damage of CSE-induced BEAS-2B cells. Collectively, JMJD3 depletion might suppress ferroptosis mediated by ACSL4 to alleviate CSE-triggered inflammation and oxidative stress in BEAS-2B cells.
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¹: Huisuo Hong and Qingqing Huang contributed equally to this work.

View full text

Dexmedetomidine preconditioning ameliorates lung injury induced by pulmonary ischemia/reperfusion by upregulating promoter histone H3K4me3 modification of KGF-2

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Antibodies and reagents

DexP alleviates I/R-induced lung injury and endothelial barrier dysfunction in mice

Discussion

Conclusion

Credit author statement

Declaration of competing interest

Acknowledgments

Biochem. Biophys. Res. Commun.

Int. J. Biochem. Cell Biol.

Redox Biol

Brain Res.

J. Surg. Res.

Biomed. Pharmacother.

Biomed. Pharmacother.

J. Surg. Res.

Inhibition of p38 MAPK mitigates lung ischemia reperfusion injury by reducing blood-air barrier hyperpermeability

Front. Pharmacol.

Novel strategies for endothelial preservation in lung transplant ischemia-reperfusion injury

Front. Physiol.

Mechanisms of lung ischemia-reperfusion injury

Curr. Opin. Organ Transplant.

Molecular targets and mechanism of action of dexmedetomidine in treatment of ischemia/reperfusion injury

Mol. Med. Rep.

Dexmedetomidine maintains its direct cardioprotective effect against ischemia/reperfusion injury in hypertensive hypertrophied myocardium

Anesth. Analg.

Effect of dexmedetomidine on lung is chemiareperfusion injury

Mol. Med. Rep.

Protective effects of keratinocyte growth factor-2 on ischemia-reperfusion-induced lung injury in rats

Am. J. Respir. Cell Mol. Biol.

Keratinocyte growth factor-2 intratracheal instillation significantly attenuates ventilator-induced lung injury in rats

J. Cell Mol. Med.

Jmjd3 contributes to the control of gene expression in LPS-activated macrophages

EMBO J.

Critical role of histone demethylase Jmjd3 in the regulation of CD4+ T-cell differentiation

Nat. Commun.