Sex-specific effects of N-acetylcysteine in neonatal rats treated with hypothermia after severe hypoxia-ischemia
Xingju Nie, Danielle W Lowe, Laura Grace Rollins, Jessica Bentzley, Jamie L Fraser, Renee Martin, Inderjit Singh, Dorothea Jenkins, Xingju Nie, Danielle W Lowe, Laura Grace Rollins, Jessica Bentzley, Jamie L Fraser, Renee Martin, Inderjit Singh, Dorothea Jenkins
Abstract
Approximately half of moderate to severely hypoxic-ischemic (HI) newborns do not respond to hypothermia, the only proven neuroprotective treatment. N-acetylcysteine (NAC), an antioxidant and glutathione precursor, shows promise for neuroprotection in combination with hypothermia, mitigating post-HI neuroinflammation due to oxidative stress. As mechanisms of HI injury and cell death differ in males and females, sex differences must be considered in translational research of neuroprotection. We assessed the potential toxicity and efficacy of NAC in combination with hypothermia, in male and female neonatal rats after severe HI injury. NAC 50mg/kg/d administered 1h after initiation of hypothermia significantly decreased iNOS expression and caspase 3 activation in the injured hemisphere versus hypothermia alone. However, only females treated with hypothermia +NAC 50mg/kg showed improvement in short-term infarct volumes compared with saline treated animals. Hypothermia alone had no effect in this severe model. When NAC was continued for 6 weeks, significant improvement in long-term neuromotor outcomes over hypothermia treatment alone was observed, controlling for sex. Antioxidants may provide insufficient neuroprotection after HI for neonatal males in the short term, while long-term therapy may benefit both sexes.
Keywords: N-acetylcysteine; Neonatal hypoxia ischemia; Neuroprotection; Redox regulation; Sex.
Copyright © 2016 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.
Figures
![Figure 1](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/4903952/bin/nihms-762644-f0002.jpg)
![Figure 2. Activated caspase 3+ cells in…](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/4903952/bin/nihms-762644-f0003.jpg)
Figure 3. PCR analysis of inflammatory mediators…
Figure 3. PCR analysis of inflammatory mediators at 24h, normalized to β-actin
Median and 95%…
Figure 4. PCR analysis of inflammatory mediators…
Figure 4. PCR analysis of inflammatory mediators at 24h by sex, normalized to β-actin
Box…
Figure 5. Infarct volumes at 48h
Median…
Figure 5. Infarct volumes at 48h
Median values are marked by bar. For the overall…
Figure 6. Infarct volumes at 48h with…
Figure 6. Infarct volumes at 48h with delayed initiation of therapy
Median values (irrespective of…
Figure 7. Functional outcomes at 7 weeks…
Figure 7. Functional outcomes at 7 weeks in controls (left panels) and in NAC treatment…
- Vitamin D improves functional outcomes in neonatal hypoxic ischemic male rats treated with N-acetylcysteine and hypothermia.Lowe DW, Fraser JL, Rollins LG, Bentzley J, Nie X, Martin R, Singh I, Jenkins D. Lowe DW, et al. Neuropharmacology. 2017 Sep 1;123:186-200. doi: 10.1016/j.neuropharm.2017.06.004. Epub 2017 Jun 6. Neuropharmacology. 2017. PMID: 28599922
- Combination of systemic hypothermia and N-acetylcysteine attenuates hypoxic-ischemic brain injury in neonatal rats.Jatana M, Singh I, Singh AK, Jenkins D. Jatana M, et al. Pediatr Res. 2006 May;59(5):684-9. doi: 10.1203/01.pdr.0000215045.91122.44. Pediatr Res. 2006. PMID: 16627882
- Neuroprotection with hypothermia and allopurinol in an animal model of hypoxic-ischemic injury: Is it a gender question?Rodríguez-Fanjul J, Durán Fernández-Feijóo C, Lopez-Abad M, Lopez Ramos MG, Balada Caballé R, Alcántara-Horillo S, Camprubí Camprubí M. Rodríguez-Fanjul J, et al. PLoS One. 2017 Sep 20;12(9):e0184643. doi: 10.1371/journal.pone.0184643. eCollection 2017. PLoS One. 2017. PMID: 28931035 Free PMC article.
- Intervention strategies for neonatal hypoxic-ischemic cerebral injury.Perlman JM. Perlman JM. Clin Ther. 2006 Sep;28(9):1353-65. doi: 10.1016/j.clinthera.2006.09.005. Clin Ther. 2006. PMID: 17062309 Review.
- Therapeutic hypothermia as a neuroprotective strategy in neonatal hypoxic-ischemic brain injury and traumatic brain injury.Ma H, Sinha B, Pandya RS, Lin N, Popp AJ, Li J, Yao J, Wang X. Ma H, et al. Curr Mol Med. 2012 Dec;12(10):1282-96. doi: 10.2174/156652412803833517. Curr Mol Med. 2012. PMID: 22834830 Free PMC article. Review.
- N-Acetylcysteine Administration Attenuates Sensorimotor Impairments Following Neonatal Hypoxic-Ischemic Brain Injury in Rats.Kesidou E, Bitsina C, Chatzisotiriou A, Theotokis P, Dandi E, Tata DA, Spandou E. Kesidou E, et al. Int J Mol Sci. 2022 Dec 19;23(24):16175. doi: 10.3390/ijms232416175. Int J Mol Sci. 2022. PMID: 36555816 Free PMC article.
- Cerebral Oxygenation and Metabolism After Hypoxia-Ischemia.Dhillon SK, Gunn ER, Lear BA, King VJ, Lear CA, Wassink G, Davidson JO, Bennet L, Gunn AJ. Dhillon SK, et al. Front Pediatr. 2022 Jul 12;10:925951. doi: 10.3389/fped.2022.925951. eCollection 2022. Front Pediatr. 2022. PMID: 35903161 Free PMC article. Review.
- Therapeutic hypothermia for the treatment of neonatal hypoxia-ischemia: sex-dependent modulation of reactive astrogliosis.Fabres RB, Nunes RR, de Medeiros de Mattos M, Andrade MKG, Martini APR, Tassinari ID, Sanches EF, de Fraga LS, Netto CA. Fabres RB, et al. Metab Brain Dis. 2022 Oct;37(7):2315-2329. doi: 10.1007/s11011-022-01030-4. Epub 2022 Jul 2. Metab Brain Dis. 2022. PMID: 35778625
- NAC and Vitamin D Improve CNS and Plasma Oxidative Stress in Neonatal HIE and Are Associated with Favorable Long-Term Outcomes.Jenkins DD, Moss HG, Brown TR, Yazdani M, Thayyil S, Montaldo P, Vento M, Kuligowski J, Wagner C, Hollis BW, Wiest DB. Jenkins DD, et al. Antioxidants (Basel). 2021 Aug 25;10(9):1344. doi: 10.3390/antiox10091344. Antioxidants (Basel). 2021. PMID: 34572976 Free PMC article.
- Mitochondrial dysfunction in perinatal asphyxia: role in pathogenesis and potential therapeutic interventions.Samaiya PK, Krishnamurthy S, Kumar A. Samaiya PK, et al. Mol Cell Biochem. 2021 Dec;476(12):4421-4434. doi: 10.1007/s11010-021-04253-8. Epub 2021 Sep 1. Mol Cell Biochem. 2021. PMID: 34472002 Review.
- Acetylcysteine / administration & dosage
- Acetylcysteine / therapeutic use*
- Animals
- Animals, Newborn
- Antioxidants / administration & dosage
- Antioxidants / therapeutic use*
- Brain / drug effects
- Brain / metabolism
- Brain / pathology
- Brain Infarction / pathology
- Brain Infarction / therapy
- Caspase 3 / metabolism
- Cell Death
- Dose-Response Relationship, Drug
- Enzyme Activation
- Female
- Hypothermia, Induced*
- Hypoxia-Ischemia, Brain / pathology
- Hypoxia-Ischemia, Brain / physiopathology
- Hypoxia-Ischemia, Brain / therapy*
- Inflammation Mediators / metabolism
- Male
- Motor Skills / drug effects
- Nitric Oxide Synthase Type II / metabolism
- Rats, Sprague-Dawley
- Sex Factors
- Time Factors
- Antioxidants
- Inflammation Mediators
- Nitric Oxide Synthase Type II
- Caspase 3
- Acetylcysteine
- Full Text Sources
- Other Literature Sources
- Medical
- Research Materials
NCBI Literature Resources
The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Unauthorized use of these marks is strictly prohibited.
National Library of Medicine
8600 Rockville Pike
Bethesda, MD 20894
![Figure 3. PCR analysis of inflammatory mediators…](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/4903952/bin/nihms-762644-f0004.jpg)
Figure 4. PCR analysis of inflammatory mediators…
Figure 4. PCR analysis of inflammatory mediators at 24h by sex, normalized to β-actin
Box…
Figure 5. Infarct volumes at 48h
Median…
Figure 5. Infarct volumes at 48h
Median values are marked by bar. For the overall…
Figure 6. Infarct volumes at 48h with…
Figure 6. Infarct volumes at 48h with delayed initiation of therapy
Median values (irrespective of…
Figure 7. Functional outcomes at 7 weeks…
Figure 7. Functional outcomes at 7 weeks in controls (left panels) and in NAC treatment…
- Vitamin D improves functional outcomes in neonatal hypoxic ischemic male rats treated with N-acetylcysteine and hypothermia.Lowe DW, Fraser JL, Rollins LG, Bentzley J, Nie X, Martin R, Singh I, Jenkins D. Lowe DW, et al. Neuropharmacology. 2017 Sep 1;123:186-200. doi: 10.1016/j.neuropharm.2017.06.004. Epub 2017 Jun 6. Neuropharmacology. 2017. PMID: 28599922
- Combination of systemic hypothermia and N-acetylcysteine attenuates hypoxic-ischemic brain injury in neonatal rats.Jatana M, Singh I, Singh AK, Jenkins D. Jatana M, et al. Pediatr Res. 2006 May;59(5):684-9. doi: 10.1203/01.pdr.0000215045.91122.44. Pediatr Res. 2006. PMID: 16627882
- Neuroprotection with hypothermia and allopurinol in an animal model of hypoxic-ischemic injury: Is it a gender question?Rodríguez-Fanjul J, Durán Fernández-Feijóo C, Lopez-Abad M, Lopez Ramos MG, Balada Caballé R, Alcántara-Horillo S, Camprubí Camprubí M. Rodríguez-Fanjul J, et al. PLoS One. 2017 Sep 20;12(9):e0184643. doi: 10.1371/journal.pone.0184643. eCollection 2017. PLoS One. 2017. PMID: 28931035 Free PMC article.
- Intervention strategies for neonatal hypoxic-ischemic cerebral injury.Perlman JM. Perlman JM. Clin Ther. 2006 Sep;28(9):1353-65. doi: 10.1016/j.clinthera.2006.09.005. Clin Ther. 2006. PMID: 17062309 Review.
- Therapeutic hypothermia as a neuroprotective strategy in neonatal hypoxic-ischemic brain injury and traumatic brain injury.Ma H, Sinha B, Pandya RS, Lin N, Popp AJ, Li J, Yao J, Wang X. Ma H, et al. Curr Mol Med. 2012 Dec;12(10):1282-96. doi: 10.2174/156652412803833517. Curr Mol Med. 2012. PMID: 22834830 Free PMC article. Review.
- N-Acetylcysteine Administration Attenuates Sensorimotor Impairments Following Neonatal Hypoxic-Ischemic Brain Injury in Rats.Kesidou E, Bitsina C, Chatzisotiriou A, Theotokis P, Dandi E, Tata DA, Spandou E. Kesidou E, et al. Int J Mol Sci. 2022 Dec 19;23(24):16175. doi: 10.3390/ijms232416175. Int J Mol Sci. 2022. PMID: 36555816 Free PMC article.
- Cerebral Oxygenation and Metabolism After Hypoxia-Ischemia.Dhillon SK, Gunn ER, Lear BA, King VJ, Lear CA, Wassink G, Davidson JO, Bennet L, Gunn AJ. Dhillon SK, et al. Front Pediatr. 2022 Jul 12;10:925951. doi: 10.3389/fped.2022.925951. eCollection 2022. Front Pediatr. 2022. PMID: 35903161 Free PMC article. Review.
- Therapeutic hypothermia for the treatment of neonatal hypoxia-ischemia: sex-dependent modulation of reactive astrogliosis.Fabres RB, Nunes RR, de Medeiros de Mattos M, Andrade MKG, Martini APR, Tassinari ID, Sanches EF, de Fraga LS, Netto CA. Fabres RB, et al. Metab Brain Dis. 2022 Oct;37(7):2315-2329. doi: 10.1007/s11011-022-01030-4. Epub 2022 Jul 2. Metab Brain Dis. 2022. PMID: 35778625
- NAC and Vitamin D Improve CNS and Plasma Oxidative Stress in Neonatal HIE and Are Associated with Favorable Long-Term Outcomes.Jenkins DD, Moss HG, Brown TR, Yazdani M, Thayyil S, Montaldo P, Vento M, Kuligowski J, Wagner C, Hollis BW, Wiest DB. Jenkins DD, et al. Antioxidants (Basel). 2021 Aug 25;10(9):1344. doi: 10.3390/antiox10091344. Antioxidants (Basel). 2021. PMID: 34572976 Free PMC article.
- Mitochondrial dysfunction in perinatal asphyxia: role in pathogenesis and potential therapeutic interventions.Samaiya PK, Krishnamurthy S, Kumar A. Samaiya PK, et al. Mol Cell Biochem. 2021 Dec;476(12):4421-4434. doi: 10.1007/s11010-021-04253-8. Epub 2021 Sep 1. Mol Cell Biochem. 2021. PMID: 34472002 Review.
- Acetylcysteine / administration & dosage
- Acetylcysteine / therapeutic use*
- Animals
- Animals, Newborn
- Antioxidants / administration & dosage
- Antioxidants / therapeutic use*
- Brain / drug effects
- Brain / metabolism
- Brain / pathology
- Brain Infarction / pathology
- Brain Infarction / therapy
- Caspase 3 / metabolism
- Cell Death
- Dose-Response Relationship, Drug
- Enzyme Activation
- Female
- Hypothermia, Induced*
- Hypoxia-Ischemia, Brain / pathology
- Hypoxia-Ischemia, Brain / physiopathology
- Hypoxia-Ischemia, Brain / therapy*
- Inflammation Mediators / metabolism
- Male
- Motor Skills / drug effects
- Nitric Oxide Synthase Type II / metabolism
- Rats, Sprague-Dawley
- Sex Factors
- Time Factors
- Antioxidants
- Inflammation Mediators
- Nitric Oxide Synthase Type II
- Caspase 3
- Acetylcysteine
- Full Text Sources
- Other Literature Sources
- Medical
- Research Materials
![Figure 4. PCR analysis of inflammatory mediators…](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/4903952/bin/nihms-762644-f0005.jpg)
![Figure 5. Infarct volumes at 48h](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/4903952/bin/nihms-762644-f0006.jpg)
![Figure 6. Infarct volumes at 48h with…](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/4903952/bin/nihms-762644-f0007.jpg)
![Figure 7. Functional outcomes at 7 weeks…](https://www.ncbi.nlm.nih.gov/pmc/articles/instance/4903952/bin/nihms-762644-f0008.jpg)
Source: PubMed