Protective role and mechanism of neural stem cells combined with edaravone in cortical neurons after oxygen-glucose deprivation/reperfusion
10.3760/cma.j.cn115354-20230428-00254
- VernacularTitle:神经干细胞联合依达拉奉对氧糖剥夺-再灌注皮层神经元的保护作用及机制探讨
- Author:
Shina SONG
1
;
Wenping DONG
;
Changxin LI
;
Jiangang DUAN
Author Information
1. 太原钢铁（集团）有限公司总医院老年内科，太原 030009
- Keywords:
Neural stem cell;
Edaravone;
Cortical neuron;
Oxygen-glucose deprivation/reperfusion
- From:
Chinese Journal of Neuromedicine
2023;22(9):918-922
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the protective role and possible mechanism of neural stem cells combined with edaravone in cortical neurons after oxygen-glucose deprivation/reperfusion (OGD-R).Methods:(1) Neural stem cells from brain tissues of SD fetal rats aged 14-16 d were cultured in vitro, and identified with Nestin, glial fibrillary acidic protein (GFAP), and microtubule-associated protein 2 (MAP2) immunofluorescent staining. Expressions of neuron-specific nuclear protein (NeuN) and β-Tubulin were detected by immunofluorescent staining in primary cortical neurons from SD rats born within 24 h. (2) Primary cortical neurons were divided into normal group (normal culture), OGD-R model group (re-oxygenated culture for 24 h after hypoxia for 1.5 h), OGD-R+neural stem cells group (re-oxygenated co-culture with cortical neurons and neural stem cells for 24 h after hypoxia for 1.5 h), OGD-R+edaravone group (re-oxygenated culture for 24 h after hypoxia for 1.5 h; 100 μmol/L edaravone before hypoxia), OGD-R+neural stem cells+edaravone group (re-oxygenated co-culture with cortical neurons and neural stem cells for 24 h after hypoxia for 1.5 h; 100 μmol/L edaravone before hypoxia); 24 h after each treatment, neuron proliferation in each group was detected by cell counting Kit 8 (CCK8), apoptosis rate was detected by flow cytometry, contents of interleukin-1β (IL-1β), and tumor necrosis factor α (TNF-α) in neuronal supernatant were detected by enzyme-linked immunosorbent assay (ELISA), and real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) and Western blotting were used to detect the mRNA and protein expressions of Notch1, Hes1 and Hes5, respectively. Results:(1) Immunofluorescent staining results showed that neural stem cells were positive for Nestin, GFAP and MAP2, and cortical neurons were positive for NeuN and β-Tubulin; all of them were successfully identified. (2) Compared with normal group, OGD-R model group, OGD-R+neural stem cell group and OGD-R+edaravone group had decreased neuron viability, increased apoptosis, increased supernatant IL-1β and TNF-α contents, and increased Notch1 mRNA and protein expressions, with significant differences ( P<0.05). Compared with OGD-R model group, OGD-R+neural stem cells+edaravone group had increased neuron viability, decreased apoptosis, decreased supernatant IL-1β and TNF-α contents, and decreased mRNA and protein expressions of Hes1 and Hes5, with significant differences ( P<0.05). Compared with the OGD-R model group, OGD-R+edaravone group and OGD-R+neural stem cell+edaravone group had significantly decreased Notch1 mRNA and protein expressions ( P<0.05). Conclusion:Combination of edaravone and neural stem cell therapy can reverse the neuronal damage caused by OGD-R, whose mechanism may be by inhibiting the expressions of inflammatory factors and key signaling molecules in Notch signaling pathway, such as Notch1, Hes1, and Hes5.
