title: Neuroprotective effect of neuroserpin in oxygen-glucose deprivation- and reoxygenation-treated rat astrocytes in vitro.
abstract: Neuroserpin (NSP) reportedly exerts neuroprotective effects in cerebral ischemic animal models and patients; however, the mechanism of protection is poorly understood. We thus attempted to confirm neuroprotective effects of NSP on astrocytes in the ischemic state and then explored the relative mechanisms. Astrocytes from neonatal rats were treated with oxygen-glucose deprivation (OGD) followed by reoxygenation (OGD/R). To confirm the neuroprotective effects of NSP, we measured the cell survival rate, relative lactate dehydrogenase (LDH) release; we also performed morphological methods, namely Hoechst 33342 staining and Annexin V assay. To explore the potential mechanisms of NSP, the release of nitric oxide (NO) and TNF-α related to NSP administration were measured by enzyme-linked immunosorbent assay. The proteins related to the NF-κB, ERK1/2, and PI3K/Akt pathways were investigated by Western blotting. To verify the cause-and-effect relationship between neuroprotection and the NF-κB pathway, a NF-κB pathway inhibitor sc3060 was employed to observe the effects of NSP-induced neuroprotection. We found that NSP significantly increased the cell survival rate and reduced LDH release in OGD/R-treated astrocytes. It also reduced NO/TNF-α release. Western blotting showed that the protein levels of p-IKKBα/β and P65 were upregulated by the OGD/R treatment and such effects were significantly inhibited by NSP administration. The NSP-induced inhibition could be significantly reversed by administration of the NF-κB pathway inhibitor sc3060, whereas, expressions of p-ERK1, p-ERK2, and p-AKT were upregulated by the OGD/R treatment; however, their levels were unchanged by NSP administration. Our results thus verified the neuroprotective effects of NSP in ischemic astrocytes. The potential mechanisms include inhibition of the release of NO/TNF-α and repression of the NF-κB signaling pathways. Our data also indicated that NSP has little influence on the MAPK and PI3K/Akt pathways.
authors: Department of Neurology, Huashan Hospital, Fudan University, Shanghai, China; Institute of Neurology, Huashan Hospital, Fudan University, Shanghai, China.
authors: Liang
authors: L
authors: Wang
authors: Department of Neurology, Huashan Hospital, Fudan University, Shanghai, China; Department of Neurology, Drum Tower Hospital, Nanjing University School of Medicine, Nanjing, China.
authors: Yang
authors: Y
authors: Zhang
authors: Department of Neurosurgery, Hamamatsu University School of Medicine, Hamamatsu-city, Japan; Department of Psychiatry, Hamamatsu University School of Medicine, Hamamatsu-city, Japan.
authors: Tetsuya
authors: T
authors: Asakawa
authors: Department of Neurology, Zhongshan Hospital, Fudan University, Shanghai, China.
authors: Wei
authors: W
authors: Li
authors: Department of Neurology, Huashan Hospital, Fudan University, Shanghai, China.
authors: Sha
authors: S
authors: Han
authors: Institute of Neurology, Huashan Hospital, Fudan University, Shanghai, China.
authors: Qinying
authors: Q
authors: Li
authors: Institute of Neurology, Huashan Hospital, Fudan University, Shanghai, China.
authors: Baoguo
authors: B
authors: Xiao
authors: Department of Neurosurgery, Hamamatsu University School of Medicine, Hamamatsu-city, Japan.
authors: Hiroki
authors: H
authors: Namba
authors: Department of Neurology, Huashan Hospital, Fudan University, Shanghai, China; Institute of Neurology, Huashan Hospital, Fudan University, Shanghai, China.
authors: Chuanzhen
authors: C
authors: Lu
authors: Department of Neurology, Huashan Hospital, Fudan University, Shanghai, China; Institute of Neurology, Huashan Hospital, Fudan University, Shanghai, China.
authors: Qiang
authors: Q
authors: Dong
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D017304:Annexin A5 / Q000737:chemistry; D017209:Apoptosis; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D001562:Benzimidazoles / Q000737:chemistry; D001786:Blood Glucose / Q000737:chemistry; D002470:Cell Survival; D002478:Cells, Cultured; D005947:Glucose / Q000737:chemistry*; D007511:Ischemia / Q000473:pathology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009479:Neuropeptides / Q000502:physiology*; D018696:Neuroprotective Agents / Q000737:chemistry*; D009569:Nitric Oxide / Q000737:chemistry; D010100:Oxygen / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015843:Serpins / Q000502:physiology*; D015398:Signal Transduction; D000097522:Neuroserpin
chemical_list: D017304:Annexin A5; D001562:Benzimidazoles; D001786:Blood Glucose; D005947:Glucose; D007770:L-Lactate Dehydrogenase; D016328:NF-kappa B; D009479:Neuropeptides; D018696:Neuroprotective Agents; D009569:Nitric Oxide; D010100:Oxygen; D010455:Peptides; D019869:Phosphatidylinositol 3-Kinases; D015843:Serpins; D000097522:Neuroserpin; C118258:SN50 peptide; C017807:bisbenzimide ethoxide trihydrochloride