Elsevier

Experimental Neurology

Volume 323, January 2020, 113066
Experimental Neurology

Research paper
Acrolein is involved in ischemic stroke-induced neurotoxicity through spermidine/spermine-N1-acetyltransferase activation

https://doi.org/10.1016/j.expneurol.2019.113066Get rights and content

Highlights

  • A deregulated acrolein metabolism is found in stroke patients.

  • Acrolein is produced resulting in brain damage in MCAO animal model.

  • Acrolein occurs through SSAT-induced polyamine oxidation by NF-kB pathway.

  • Acrolein elicits a vicious cycling of oxidative stress resulting in neurotoxicity.

  • N-acetylcysteine prevents OGD-induced neurotoxicity by scavenging acrolein.

Abstract

Background and purpose

Ischemic stroke is the most common type of cerebrovascular event and is responsible for approximately 85% of all strokes in Taiwan. Neurons contain high concentrations of polyamines, which are prone to various pathological states in the brain and are perturbed after cerebral ischemia. Acrolein, an α,β-unsaturated aldehyde, has been suggested as the primary culprit of neuronal damage in stroke patients. However, the mechanism by which acrolein induces neuronal damage during ischemic stroke is not clear.

Methods

Urinary 3-hydroxypropyl mercapturic acid (3-HPMA), an acrolein-glutathione (GSH) metabolite, plasma acrolein-protein conjugates (Acr-PC) and plasma GSH levels were analyzed to correlate disease severity and prognosis of stroke patients compared with control subjects. In vivo middle cerebral artery occlusion (MCAO) animal models and an in vitro oxygen glucose deprivation (OGD) stroke model were used to investigate the mechanisms of acrolein-induced neuronal damage.

Results

A deregulated acrolein metabolism, including significantly increased plasma Acr-PC levels, decreased urinary 3-HPMA levels and decreased plasma GSH levels, was found in stroke patients compared to control subjects. We further observed that acrolein was produced during ischemia resulting in brain damage in in vivo MCAO animal model. The induction of acrolein in neuronal cells during OGD occurred due to the increased expression of spermidine/spermine N1-acetyltransferase (SSAT) by NF-kB pathway activation. In addition, acrolein elicited a vicious cycling of oxidative stress resulting in neurotoxicity. Finally, N-acetylcysteine effectively prevented OGD-induced neurotoxicity by scavenging acrolein.

Conclusion

Overall, our current results demonstrate that acrolein is a culprit of neuronal damage through GSH depletion in stroke patients. The mechanism underlying the role of acrolein in stroke-related neuronal damage occurs through SSAT-induced polyamine oxidation by NF-kB pathway activation. These results provide a novel mechanism of neurotoxicity in stroke patients, aid in the development of neutralizing or preventive measures, and further our understanding of neural protection.

Introduction

Ischemic stroke, characterized by the disturbance of the blood supply to the brain, is the most common type of cerebrovascular event and is responsible for approximately 85% of all strokes (Flynn et al., 2008; Mathers et al., 2009). Identifying reliable biomarkers present during the early phases of a stroke remains a challenge (Makris et al., 2018). Polyamines (putrescine, spermidine and spermine) are ubiquitous cellular polycations essential for optimal rates of cell growth and differentiation (Pegg, 2009). The polyamine system is prone to various pathological states in the brain and is perturbed after cerebral ischemia (Baskaya et al., 1997; Baskaya et al., 1996; Seiler, 2000). Spermidine/ Spermine N1-acetyltransferase (SSAT) is the rate-limiting enzyme in polyamine catabolism that acts through the acetylation of spermidine and spermine (Pegg, 2008). Previous studies have shown that the transcription and enzymatic activity of SSAT increase in response to cerebral ischemia (Nagesh Babu et al., 2001; Zoli et al., 1996).

Acrolein, an α,β-unsaturated aldehyde, can be formed endogenously by lipid peroxidation or amino acid metabolism, including polyamine oxidation (Stevens and Maier, 2008). Recent studies have shown that acrolein is predominantly formed from 3-aminopropanal (NH2[CH2]2CHO) produced effectively from spermine by spermine oxidase (SMO) and less effectively from 3-acetamidopropanal (CH3CONH[CH2]2CHO), which is produced from spermine and spermidine by SSAT and acetylpolyamine oxidase (AcPAO) (Igarashi and Kashiwagi, 2011; Kimes and Morris, 1971; Saiki et al., 2011; Wang and Casero Jr., 2006). Additionally, acrolein has been reported to be more toxic than H2O2 (Sharmin et al., 2001) and slightly more toxic than ·OH (Yoshida et al., 2009) in cell culture systems due to its high reactivity with various cellular components such as proteins, which results in protein dysfunction. Acrolein reacts with lysine residues within proteins to form Nε-(3-formyl-3,4-dehydropiperidino)lysine (FDP-lysine) and (Nε-(3-methylpyridium)lysine] (MP-lysine) (Furuhata et al., 2003; Stevens and Maier, 2008; Uchida et al., 1998), which has been used to detect acrolein levels in human tissues and serum (Calingasan et al., 1999; Igarashi and Kashiwagi, 2011). Previous studies have shown that levels of acrolein-protein conjugates (Acr-PC) in plasma are associated with disease severity in stroke patients (Igarashi and Kashiwagi, 2011; Saiki et al., 2009; Tomitori et al., 2005; Yoshida et al., 2010; Yoshida et al., 2011). Reduction of acrolein levels has been shown to reduce infarction size and protect against neuronal damage in animal experiments (Saiki et al., 2009; Suabjakyong et al., 2015). Therefore, acrolein has been suggested as a primary culprit of neuronal damage in stroke patients. However, the mechanism by which acrolein induces neuronal damage is not clear.

The governing pathobiology of ischemic brain stroke initially unfolds due to a focal deficit in metabolism driven primarily by oxygen and glucose deprivation (OGD); most ischemic strokes (~80%) occur in the middle cerebral artery (MCA). MCA occlusion (MCAO) in animals is commonly used to model focal brain ischemia in humans (Carmichael, 2005; Durukan and Tatlisumak, 2007; Taniguchi and Andreasson, 2008) and OGD is widely used as an in vitro model for stroke, due to similarities with in vivo models of brain ischemia (Tasca et al., 2015). To clarify the role of acrolein in neuronal damage during stroke, we collected blood and urine samples from 49 stroke patients within 24 h (day 1) after stroke and from 99 healthy subjects. Urinary 3-HPMA, an acrolein-glutathione (GSH) metabolite, plasma acrolein-protein conjugates (Acr-PC) and plasma GSH levels were analyzed to correlate disease severity and prognosis of stroke patients. Furthermore, the mechanisms by which acrolein induces neuronal damage were investigated using MCAO animal models and an in vitro OGD stroke model.

Section snippets

Subjects and collection of blood and urine

Baseline sociodemographic variables are shown in Table 1. From the 148 patients recruited, 49 patients were gender- and smoking history-matched stroke patients, and 99 patients were gender- and smoking history-matched healthy control subjects without any history of dementia. All stroke subjects were clinically followed for 1–2 years. (Table 1). The demographic data for all patients are shown in Table 1. Of the stroke patients, 81% (39/49) were diagnosed with a minor stroke (NIHSS <5), and the

Subjects and collection of blood and urine

Volunteers recruited in this study included 99 control subjects without any history of dementia and 49 stroke subjects as described in Supplementary Methods. Informed consent was given by each participant or his (or her) relative, and our study protocol was approved by the Institutional Review Board of Taipei Veterans General Hospital (IRB# 2017–11-003 BCE). The experiments were conducted in accordance with the Declaration of Helsinki principles.

Procedures for middle cerebral artery occlusion (MCAO)

All animal experiments were approved by the

Discussion

Acrolein, an α,β-unsaturated aldehyde, has been suggested as a primary culprit of neuronal damage in stroke patients (Igarashi and Kashiwagi, 2011; Saiki et al., 2009; Tomitori et al., 2005; Yoshida et al., 2010; Yoshida et al., 2011). However, the mechanism by which acrolein induced neuronal damages during ischemic stroke is not clear. In this study, a deregulated acrolein metabolism, including significantly increased plasma Acr-PC levels, decreased urinary 3-HPMA levels and decreased plasma

Acknowledgements

We thank Ms. Ching-Wei Lin, Ms. Shao-Yu Lin, Ms. Shiu-Hao Tai, Ms. Hui-Chi Huang, and Ms. Jui-Yao Tsai from Department of Nursing, Taipei Veterans General Hospital, Taipei, Taiwan, ROC for their great help with collection of clinical samples.

Sources of funding

This work was supported by Ministry of Science and Technology, Taiwan under [107-2321-B-010-006 (H-T Wang); 107-2320-B-010-018 (H-T Wang)].

Declaration of Competing Interest

The authors have no actual or potential conflicts of interest.

References (53)

  • G. Nagesh Babu et al.

    Spermidine/spermine N1-acetyl transferase activity in rat brain following transient focal cerebral ischemia and reperfusion

    Neurosci. Lett.

    (2001)
  • R. Saiki et al.

    Brain infarction correlates more closely with acrolein than with reactive oxygen species

    Biochem. Biophys. Res. Commun.

    (2011)
  • S. Sharmin et al.

    Polyamine cytotoxicity in the presence of bovine serum amine oxidase

    Biochem. Biophys. Res. Commun.

    (2001)
  • J. Tong et al.

    Do glutathione levels decline in aging human brain?

    Free Radic. Biol. Med.

    (2016)
  • K. Uchida

    Current status of acrolein as a lipid peroxidation product

    Trends Cardiovasc. Med.

    (1999)
  • K. Uchida et al.

    Acrolein is a product of lipid peroxidation reaction. Formation of free acrolein and its conjugate with lysine residues in oxidized low density lipoproteins

    J. Biol. Chem.

    (1998)
  • E.S. Weiss et al.

    Alpha II-spectrin breakdown products serve as novel markers of brain injury severity in a canine model of hypothermic circulatory arrest

    Ann. Thorac. Surg.

    (2009)
  • M. Yoshida et al.

    Acrolein toxicity: comparison with reactive oxygen species

    Biochem. Biophys. Res. Commun.

    (2009)
  • M. Yoshida et al.

    Identification of acrolein-conjugated protein in plasma of patients with brain infarction

    Biochem. Biophys. Res. Commun.

    (2010)
  • M. Yoshida et al.

    Relationship between metabolic disorders and relative risk values of brain infarction estimated by protein-conjugated acrolein, IL-6 and CRP together with age

    Clin. Chim. Acta

    (2011)
  • M. Zoli et al.

    Spermidine/spermine N1-acetyltransferase mRNA levels show marked and region-specific changes in the early phase after transient forebrain ischemia

    Brain Res. Mol. Brain Res.

    (1996)
  • J. Astrup et al.

    Thresholds in cerebral ischemia - the ischemic penumbra

    Stroke

    (1981)
  • N. Babbar et al.

    Induction of spermidine/spermine N1-acetyltransferase (SSAT) by aspirin in Caco-2 colon cancer cells

    Biochem. J.

    (2006)
  • M.K. Baskaya et al.

    Regional activity of ornithine decarboxylase and edema formation after traumatic brain injury

    Neurosurgery

    (1996)
  • N.Y. Calingasan et al.

    Protein-bound acrolein: a novel marker of oxidative stress in Alzheimer’s disease

    J. Neurochem.

    (1999)
  • S.G. Carmella et al.

    Quantitation of acrolein-derived (3-hydroxypropyl)mercapturic acid in human urine by liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry: effects of cigarette smoking

    Chem. Res. Toxicol.

    (2007)
  • Cited by (40)

    • Association between acrolein exposure and respiratory hazards: A systematic review and meta-analysis

      2023, Atmospheric Pollution Research
      Citation Excerpt :

      Acrolein is a major component of cigarette smoke and kitchen fumes (Crowley et al., 2022). It is a highly active unsaturated aldehyde, which is common in the environment, food, and water pollutants (Zhang and Schwab, 2022), and can also be formed when wood, oil, and plastic are burned (Liu et al., 2020a); (Shafie et al., 2021). Acrolein is also a powerful and highly reactive electronphile, which can maintain its activity for several days in vivo (Pradipta and Tanaka, 2021).

    • Food additive octyl gallate eliminates acrolein and inhibits bacterial growth in oil-rich food

      2022, Food Chemistry
      Citation Excerpt :

      ACR can cause oxidative, DNA, and mitochondrial damages and exacerbate cell apoptosis (Jiang et al., 2022). Accumulation of ACR in vivo has toxic effects on the respiratory (Ong, Henry, & Burcham, 2012), cardiovascular (Conklin et al., 2006), nervous (Liu et al., 2020), and digestive (Liu, et al., 2020) systems, which are associated with various diseases such as diabetes, atherosclerosis, cardiovascular disease, Alzheimer’s disease, and Parkinson’s disease (Ambaw, Zheng, Tambe, Strathearn, & Shi, 2018; Tsou et al., 2017). The United States Environmental Protection Agency (EPA) classified ACR as a high-priority air toxicant due to its high toxicity (Robert, Giffe, Jayme, & Raymond, 2017).

    • Electrophile versus oxidant modification of cysteine residues: Kinetics as a key driver of protein modification

      2022, Archives of Biochemistry and Biophysics
      Citation Excerpt :

      Acrolein is the smallest and most reactive alpha,beta-unsaturated carbonyl compound. Exposure to this toxic chemical is widespread since acrolein is endogenously produced during lipid peroxidation [4] and spermine metabolism [5]. Acrolein is also a combustion product of organic materials and therefore present in oils heated to high temperatures, cigarette smoke and automobile exhausts [6].

    View all citing articles on Scopus
    1

    Equal contribution

    View full text