Biphasic activation of nuclear factor-κB and expression of p65 and c-Rel following traumatic neuronal injury

Zhang,Huasheng; Zhang,Dingding; Li,Hua; Yan,Huiying; Zhang,Zihuan; Zhou,Chenhui; Chen,Qiang; Ye,Zhennan; Hang,Chunhua

doi:10.3892/ijmm.2018.3567

Biphasic activation of nuclear factor-κB and expression of p65 and c-Rel following traumatic neuronal injury

Authors:
- Huasheng Zhang
- Dingding Zhang
- Hua Li
- Huiying Yan
- Zihuan Zhang
- Chenhui Zhou
- Qiang Chen
- Zhennan Ye
- Chunhua Hang
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Affiliations: Department of Neurosurgery, Jinling Hospital, School of Medicine, Nanjing University, Nanjing, Jiangsu 210002, P.R. China, Department of Neurosurgery, Zhongdu Hospital, Bengbu, Anhui 233004, P.R. China, Department of Neurosurgery, Southern Medical University (Guangzhou), Jinling Hospital, Nanjing, Jiangsu 210002, P.R. China
Published online on: March 14, 2018 https://doi.org/10.3892/ijmm.2018.3567
Pages: 3203-3210
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The transcription factor nuclear factor-κB (NF-κB) has been shown to function as a key regulator of cell death or survival in neuronal cells. Previous studies indicate that the biphasic activation of NF-κB occurs following experimental neonatal hypoxia-ischemia and subarachnoid hemorrhage. However, the comprehensive understanding of NF-κB activity following traumatic brain injury (TBI) is incomplete. In the current study, an in vitro model of TBI was designed to investigate the NF-κB activity and expression of p65 and c-Rel subunits following traumatic neuronal injury. Primary cultured neurons were assigned to control and transected groups. NF-κB activity was detected by electrophoretic mobility shift assay. Western blotting and immunofluorescence were used to investigate the expression and distribution of p65 and c-Rel. Reverse transcription-quantitative polymerase chain reaction was performed to assess the downstream genes of NF-κB. Lactate dehydrogenase (LDH) quantification and trypan blue staining were used to estimate the neuronal injury. Double peaks of elevated NF-κB activity were observed at 1 and 24 h following transection. The expression levels of downstream genes exhibited similar changes. The protein levels of p65 also presented double peaks while c-Rel was elevated significantly in the late stage. The results of the trypan blue staining and LDH leakage assays indicated there was no sustained neuronal injury during the late peak of NF-κB activity. In conclusion, biphasic activation of NF-κB is induced following experimental traumatic neuronal injury. The elevation of p65 and c-Rel levels at different time periods suggests that within a single neuron, NF-κB may participate in different pathophysiological processes.

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