Abstract
Background
Magnesium sulfate (MgSO4) is utilized for fetal neuroprotection in preterm birth but its mechanism of action is still poorly understood. P2X7 receptor (P2X7R) is required for secretion of IL-1β, and can be blocked by divalent cations such as magnesium (Mg) and its own antagonist, Brilliant Blue G (BBG). We sought to determine whether during inflammation MgSO4 can block endothelial IL-1β secretion, using an in-vitro model.
Methods
Human umbilical vein endothelial cell (HUVEC) cultures were treated with varying doses of LPS, 2′(3)-Ο-(4-Benzoylbenzoyl) adenosine-5′-triphosphate (BzATP), BBG and MgSO4 for 3- or 24 h. We determined cell cytotoxicity, apoptosis, IL-1β mRNA expression, IL-1β production and secretion and P2X7R expression on HUVECs.
Results
We demonstrated that MgSO4 is efficacious in blocking IL-1β-mediated-inflammation in HUVECs, at both the initiation and propagation phases of inflammation. MgSO4 exerts these anti-inflammatory effects via downregulation of P2X7Rs on HUVECs.
Conclusion
LPS-exposure increases IL-1β production and secretion in HUVECs, which is further intensified by P2X7R agonist, BzATP while MgSO4 inhibits IL-1β in both presence and absence of BzATP. This effect is similar to the results of P2X7R antagonist, BBG, suggesting that the anti-inflammatory effects of MgSO4 is through P2X7R.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Burd, I., Balakrishnan, B. & Kannan, S. Models of fetal brain injury, intrauterine inflammation, and preterm birth. Am. J. Reprod. Immunol. 67, 287–294 (2012).
Zarrei, M. et al. De novo and rare inherited copy-number variations in the hemiplegic form of cerebral palsy. Genet Med. 20, 172–180 (2018)
Kuban, K. C. et al. systemic inflammation and cerebral palsy risk in extremely preterm infants. J. Child. Neurol. 29, 1692–1698 (2014).
Cordeiro, C. N., Tsimis, M. & Burd, I. Infections and brain development. Obstet. Gynecol. Surv. 70, 644–655 (2015).
Leitner, K. et al. IL-1 receptor blockade prevents fetal cortical brain injury but not preterm birth in a mouse model of inflammation-induced preterm birth and perinatal brain injury. Am. J. Reprod. Immunol. 71, 418–426 (2014).
Rouse, D. et al. A randomized, controlled trial of magnesium sulfate for the prevention of cerebral palsy. N. Engl. J. Med. 359, 895–905 (2008).
Stigson, L. & Kjellmer, I. Serum levels of magnesium at birth related to complications of immaturity. Acta Paediatr. 86, 991–994 (1997).
Hirtz, D. G. & Nelson, K. Magnesium sulfate and cerebral palsy in premature infants. Curr. Opin. Pediatr. 10, 131–137 (1998).
Almousa, L. A., Salter, A. M. & Langley-Evans, S. C. Varying magnesium concentration elicits changes in inflammatory response in human umbilical vein endothelial cells (HUVECs). Magnes. Res. 31, 99–109 (2018).
Pappas, A. et al. Chorioamnionitis and early childhood outcomes among extremely low-gestational-age neonates. JAMA Pediatr. 168, 137–147 (2014).
Edwards, J. M., Edwards, L. E., Swamy, G. K. & Grotegut, C. A. Magnesium sulfate for neuroprotection in the setting of chorioamnionitis. J. Matern. Fetal. Neonatal Med. 31, 1156–1160 (2018).
Kim, C. J. et al. Acute chorioamnionitis and funisitis: definition, pathologic features, and clinical significance. Am. J. Obstet. Gynecol. 213, S29–S52 (2015).
Di Virgilio, F., Dal Ben, D., Sarti, A. C., Giuliani, A. L. & Falzoni, S. The P2X7 receptor in infection and inflammation. Immunity 47, 15–31 (2017).
Bartlett, R., Stokes, L. & Sluyter, R. The P2X7 receptor channel: recent developments and the use of P2X7 antagonists in models of disease. Pharm. Rev. 66, 638–675 (2014).
Wilson, H. L. et al. P2X receptor characterization and IL-1/IL-1Ra release from human endothelial cells. Br. J. Pharm. 151, 115–127 (2007).
Tsimis, M. E. et al. P2X7 receptor blockade prevents preterm birth and perinatal brain injury in a mouse model of intrauterine inflammation. Biol. Reprod. 97, 230–239 (2017).
Younge, N. et al. Survival and neurodevelopmental outcomes among periviable infants. N. Engl. J. Med. 376, 617–628 (2017).
Durkin, M. S. et al. Prevalence of cerebral palsy among 8-year-old children in 2010 and preliminary evidence of trends in its relationship to low birthweight. Paediatr. Perinat. Epidemiol. 30, 496–510 (2016).
Brookfield, K. F., Elkomy, M., Su, F., Drover, D. R. & Carvalho, B. Optimization of maternal magnesium sulfate administration for fetal neuroprotection: application of a prospectively constructed pharmacokinetic model to the BEAM cohort. J. Clin. Pharm. 57, 1419–1424 (2017).
Edwards, J. M., Edwards, L. E., Swamy, G. K. & Grotegut, C. A. Effect of cord blood magnesium level at birth on non-neurologic neonatal outcomes. Am. J. Perinatol. 36, 3–7 (2018).
Park, J. S., Park, C. W., Lockwood, C. J. & Norwitz, E. R. Role of cytokines in preterm labor and birth. Minerva Ginecol. 57, 349–366 (2005).
D’Alquen, D. et al. Activation of umbilical cord endothelial cells and fetal inflammatory response in preterm infants with chorioamnionitis and funisitis. Pediatr. Res. 57, 263–269 (2005).
Xing, Y. L. et al. Protocatechuic aldehyde inhibits lipopolysaccharide-induced human umbilical vein endothelial cell apoptosis via regulation of caspase-3. Phytother. Res. 26, 1334–1341 (2012).
Lee, T. H., Chang, J. & Kim, B. M. Saikosaponin C inhibits lipopolysaccharide-induced apoptosis by suppressing caspase-3 activation and subsequent degradation of focal adhesion kinase in human umbilical vein endothelial cells. Biochem. Biophys. Res. Commun. 445, 615–621 (2014).
The American College of Obstetricians and Gynecologists Committee on Obstetric Practice Society for Maternal-Fetal Medicine. 2016 Committee Opinion No. 652: magnesium sulfate use in obstetrics. Obstet. Gynecol. 127, e52–e53 (2016).
Guzman-Aranguez, A., Perez de Lara, M. J. & Pintor, J. Hyperosmotic stress induces ATP release and changes in P2X7 receptor levels in human corneal and conjunctival epithelial cells. Purinergic Signal. 13, 249–258 (2017).
Maier, J. A., Malpuech-Brugere, C., Zimowska, W., Rayssiguier, Y. & Mazur, A. Low magnesium promotes endothelial cell dysfunction: implications for atherosclerosis, inflammation and thrombosis. Biochim. Biophys. Acta 1689, 13–21 (2004).
Nakagawa, M., Oono, H. & Nishio, A. Enhanced production of IL-1beta and IL-6 following endotoxin challenge in rats with dietary magnesium deficiency. J. Vet. Med. Sci. 63, 467–469 (2001).
Rochelson, B., Dowling, O., Schwartz, N. & Metz, C. N. Magnesium sulfate suppresses inflammatory responses by human umbilical vein endothelial cells (HuVECs) through the NFkappaB pathway. J. Reprod. Immunol. 73, 101–107 (2007).
Burd, I., Breen, K., Friedman, A., Chai, J. & Elovitz, M. A. Magnesium sulfate reduces inflammation-associated brain injury in fetal mice. Am. J. Obstet. Gynecol. 202, 292 e291–292 e299 (2010).
Fujiwara, M. et al. Species difference in sensitivity of human and mouse P2X7 receptors to inhibitory effects of divalent metal cations. Biol. Pharm. Bull. 40, 375–380 (2017).
Virginio, C., Church, D., North, R. A. & Surprenant, A. Effects of divalent cations, protons and calmidazolium at the rat P2X7 receptor. Neuropharmacology 36, 1285–1294 (1997).
Torigoe, G. et al. PYK2 mediates BzATP-induced extracellular matrix proteins synthesis. Biochem. Biophys. Res. Commun. 494, 663–667 (2017).
Jiang, L. H., Mackenzie, A. B., North, R. A. & Surprenant, A. Brilliant blue G selectively blocks ATP-gated rat P2X(7) receptors. Mol. Pharm. 58, 82–88 (2000).
Beigi, R. D., Kertesy, S. B., Aquilina, G. & Dubyak, G. R. Oxidized ATP (oATP) attenuates proinflammatory signaling via P2 receptor-independent mechanisms. Br. J. Pharm. 140, 507–519 (2003).
Valdecantos, P., Briones, R., Moya, P., Germain, A. & Huidobro-Toro, J. P. Pharmacological identification of P2X1, P2X4 and P2X7 nucleotide receptors in the smooth muscles of human umbilical cord and chorionic blood vessels. Placenta 24, 17–26 (2003).
Green, J. P. et al. Atheroprone flow activates inflammation via endothelial ATP-dependent P2X7-p38 signalling. Cardiovasc. Res. 114, 324–335 (2018).
Albalawi, F. et al. The P2X7 receptor primes IL-1beta and the NLRP3 inflammasome in astrocytes exposed to mechanical strain. Front. Cell Neurosci. 11, 227 (2017).
Jiang, L. H. Inhibition of P2X(7) receptors by divalent cations: old action and new insight. Eur. Biophys. J. 38, 339–346 (2009).
Acknowledgements
This study was supported by the Integrated Research Center for Fetal Medicine Fund.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Presented orally at the 65th Annual Scientific Meeting of the Society of Reproductive Immunology, San Diego, CA, March 6−10, 2018.
Supplementary information
Rights and permissions
About this article
Cite this article
Ozen, M., Xie, H., Shin, N. et al. Magnesium sulfate inhibits inflammation through P2X7 receptors in human umbilical vein endothelial cells. Pediatr Res 87, 463–471 (2020). https://doi.org/10.1038/s41390-019-0557-7
Received:
Revised:
Accepted:
Published:
Version of record:
Issue date:
DOI: https://doi.org/10.1038/s41390-019-0557-7
This article is cited by
-
Magnesium sulfate in oxidative stress-associated pathologies: clinical, cellular, and molecular perspectives
Biophysical Reviews (2025)
-
Assessment of drugs administered in the Middle East as part of the COVID-19 management protocols
Inflammopharmacology (2022)
-
Effect of infusion irrigation with different irrigating solutions on transient receptor potential vanilloid 5 and intra-articular inflammation in a post-traumatic osteoarthritis rabbit model
European Journal of Medical Research (2021)
