Effect of neuroserpin in a neonatal hypoxic-ischemic injury model ex vivo

Abstract

Hypoxia-ischemia (HI) occurring in immature brains stimulates the expression of tissue-type plasminogen activator (tPA). Neuroserpin is a selected inhibitor of tPA in the central nerves system. However, the role that neuroserpin plays and the possible mechanisms involved during neonatal HI are poorly defi ned. In this study, an oxygen-glucose deprivation and reoxygenation (OGD/R) model was generated with cultured rat cortical neurons mimicking neonatal HI injury ex vivo, and an acute neuronal excitatory injury was induced by exposure to a high concentration of N-methyl-D-aspartic acid (NMDA). Cells received either neuroserpin or MK-801, an antagonist of the NMDA receptor, during OGD/R, and were incubated with or without neuroserpin after NMDA exposure. Cell viability and morphology were detected by a Cell Counting Kit-8 and immunohistochemical staining, respectively. TPA expression and activity were also assessed. We found that MK-801 alleviated injuries induced by OGD/R, suggesting an excitatory damage involvement. Neuroserpin provided a dosedependent neuroprotective eff ect in both OGD/R and acute excitatory injuries by inhibiting the activity of tPA, without aff ecting neuronal tPA expression. Neuroserpin protected neurons against OGD/R even after a delayed administration of 3h. Collectively, our data indicate that neuroserpin protects neurons against OGD/R. mainly by inhibiting tPA-mediated acute neuronal excitotoxicity.

Neonatal Hypoxia Ischaemia: Mechanisms, Models, and Therapeutic Challenges

“…Further, neuroserpin is protective in an in vitro model of HI, OGD (Wu et al, 2010; Rodríguez-González et al, 2011a; Ma et al, 2012b). Both neurons and astrocytes cultured in OGD undergo less apoptosis and do not exhibit damaged neural processes when exogenous neuroserpin is administered following OGD (Rodríguez-González et al, 2011a; Ma et al, 2012b). Hippocampal neurons from both wild-type and tPA knockout mice were protected from OGD, plasmin-mediated cell death and kainic acid by neuroserpin administration (Wu et al, 2010), demonstrating that neuroserpin’s neuroprotective role is not exclusively dependent on tPA inhibition.…”

“…Exogenous neuroserpin administration in vitro decreases apoptosis caused by tPA, NMDA, kainic acid, and OGD (Lebeurrier et al, 2005a,b; Mohsenifar et al, 2007; Lee et al, 2008; Wu et al, 2010; Rodríguez-González et al, 2011a; Ma et al, 2012b). In addition, aberrant initiation of neuroserpin expression in cancer cells preserves the tumor and has been linked with increased treatment-resistance in prostate cancer (Chang et al, 2000; Hasumi et al, 2005; Valiente et al, 2014).…”

“…Administration of neuroserpin protects mouse cortical neurons in vitro against NMDA-induced excitotoxicity, but not AMPA-induced excitotoxicity (Lebeurrier et al, 2005a,b; Ma et al, 2012b). Further, neuroserpin is protective in an in vitro model of HI, OGD (Wu et al, 2010; Rodríguez-González et al, 2011a; Ma et al, 2012b).…”

“…Administration of neuroserpin protects mouse cortical neurons in vitro against NMDA-induced excitotoxicity, but not AMPA-induced excitotoxicity (Lebeurrier et al, 2005a,b; Ma et al, 2012b). Further, neuroserpin is protective in an in vitro model of HI, OGD (Wu et al, 2010; Rodríguez-González et al, 2011a; Ma et al, 2012b). Both neurons and astrocytes cultured in OGD undergo less apoptosis and do not exhibit damaged neural processes when exogenous neuroserpin is administered following OGD (Rodríguez-González et al, 2011a; Ma et al, 2012b).…”

Neuroserpin Protects Rat Neurons and Microglia-Mediated Inflammatory Response Against Oxygen-Glucose Deprivation- and Reoxygenation Treatments in an In Vitro Study

“…The neuroprotective role of NSP has received widespread attention, although these mechanisms have been poorly understood. NSP demonstrates widespread neuroprotective effects not only on stroke but also on neurodegenerative disorders [2][3][4][5] in both patients [6] and animal models [1,7,8]. Recently, the neuroprotective role of NSP on ischemic/hypoxia cerebral cells has been increasingly investigated.…”