Recombinant human brain-derived neurotrophic factor prevents neuronal apoptosis in a novel in vitro model of subarachnoid hemorrhage

Abstract

Subarachnoid hemorrhage (SAH) is a hemorrhagic stroke with high mortality and morbidity. An animal model for SAH was established by directly injecting a hemolysate into the subarachnoid space of rats or mice. However, the in vitro applications of the hemolysate SAH model have not been reported, and the mechanisms remain unclear. In this study, we established an in vitro SAH model by treating cortical pyramidal neurons with hemolysate. Using this model, we assessed the effects of recombinant human brain-derived neurotrophic factor (rhBDNF) on hemolysate-induced cell death and related mechanisms. Cortical neurons were treated with 10 ng/mL or 100 ng/mL rhBDNF prior to application of hemolysate. Hemolysate treatment markedly increased cell loss, triggered apoptosis, and promoted the expression of caspase-8, caspase-9, and cleaved caspase-3. rhBDNF significantly inhibited hemolysate-induced cell loss, neuronal apoptosis, and expression of caspase-8, caspase-9, and cleaved caspase-3. Our data revealed a previously unrecognized protective activity of rhBDNF against hemolysate-induced cell death, potentially via regulation of caspase-9-, caspase-8-, and cleaved caspase-3-related apoptosis. This study implicates that hemolysate-induced cortical neuron death represents an important in vitro model of SAH.

TAT-HSP27 Peptide Improves Neurologic Deficits via Reducing Apoptosis After Experimental Subarachnoid Hemorrhage

“…The N-terminal region of HSP27 proved to be a necessary domain for neuroprotection in vitro ischemia (Stetler et al, 2008 ). So, we synthesized peptides from the N-terminal region of HSP27 and found that the HSP27 65−90 peptide reduces cortical neuronal death in an in vitro hemolysate-damaged cortical neuron model (Li et al, 2017 ), which contains multiple components and mimics the pathophysiological scenario of SAH observed in vivo (Zhou et al, 2007 ). Generally, it is believed that SAH causes neuronal death in the basal cortex exposed to bloody CSF (Park et al, 2004 ).…”

“…Briefly, the dissociated neurons (10 6 cells) were plated in 60 mm dish with neurobasal medium [2% B27 (Gibco), 1% L-glutamine (Gibco), 0.3% D-glucose (Sigma), and 1% fetal bovine serum (Gibco)] and fed with fresh medium every 3 days. Hemolysate-induced cortical neuron death model was produced as described previously (Li et al, 2017 ). Briefly, hemolysate was prepared from mouse arterial whole blood by freezing and stored at −80°C.…”

Biological Effects and Mechanisms of Caspases in Early Brain Injury after Subarachnoid Hemorrhage

“…According to emerging evidence, hemolysate induces the upregulation of caspase-8 expression levels in SAH, suggesting that death receptor-related pathways are involved in hemolytic substance-induced apoptosis in cortical neurons [ 51 ]. Caspase-8 and FADD are associated with the nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain containing 3 (NLRP3) inflammasome, resulting in gasdermin D- (GSDMD-) mediated cell death in the form of pyroptosis [ 1 , 52 ].…”

Exogenous brain‑derived neurotrophic factor attenuates neuronal apoptosis and neurological deficits after subarachnoid hemorrhage in rats

“…Expression of mature BDNF was found to be decreased at 4, 48 and 72 h, and increased at 5 and 7 days post-SAH in a blood injection SAH model (10–12). Furthermore, recombinant human BDNF inhibited hemolysate-induced neuron death in an in vitro model of SAH (13). However, the role of BDNF in SAH-induced neuronal apoptosis and neurological deficits is unclear.…”