In gratitude of the loyal support of our members, the CNS is offering complimentary 2021 Annual Meeting registration to all members! Learn more.

  • Biphasic Expression of NF-kappaB in Experimental Subarachnoid Hemorrhage in vivo and vitro

    Final Number:
    349

    Authors:
    Wanchun You MD; Mengliang Zhou; Wei Li M.D.; Zong Zhuang; Xiangjun JI MD; Yong Tang; Jixin Shi MD, PhD

    Study Design:
    Laboratory Investigation

    Subject Category:

    Meeting: Congress of Neurological Surgeons 2012 Annual Meeting

    Introduction: It has been demonstrated that NF-kappaB (NF-kappaB) as a well-known transcription factor is activated after subarachnoid hemorrhage (SAH) onset. However, the existent researches only sketched the role of NF-kappaB in early brain injury after SAH, and the panoramic view of NF-kappaB activation remains obscure. In the current study, we aim to investigate the phases of NF-kappaB activation in vitro and vivo experiments and the downstream gene expression for further studies.

    Methods: Cultured neurons were treated with hemoglobin to produce the neuron SAH injury model in vitro. In the present study, the cultured neurons were divided into 7 groups: control groups, without any special treatment; 1h, 3h, 6h, 12h, 24h, 48h groups of hemoglobin injury model. One-hemorrhage SAH model in New Zealand rabbits was induced with the cisterna magna arterial blood injection methods. Forty New Zealand rabbits were signed into 6 groups (4 or 8 rabbits per group): control groups, just experience the operation procedures without blood injection; day 1, 3, 6, 10, 14 groups of SAH model. Electrophoretic mobility shift assay (EMSA) was performed to access the NF-kappaB activation both in vivo and vitro. Immunohistochemistry was used to detect the location of activated NF-kappaB, Real-time PCR were performed to assess the downstream gene expression of NF-kappaB respectively. Nissl staining and LDH quantification were used to distinguish the neuron injury in vivo and vitro respectively.

    Results: NF-kappaB DNA binding activity showed dual elevated phases after SAH in rabbits(day 1 to day 3 and day 10 groups). Meanwhile, in cultured hemoglobin-induced neuron injury model, NF-kappaB DNA binding activity showed significantly increase and biphasic peaks (1h and 24h groups) after exposed to hemoglobin. The down-stream gene expression showed an accordant peak phases detected by real-time PCR. The Nissl staining and LDH quantification results suggested neurons damage role for the early peak and no deteriorated effect for the late peak.

    Conclusions: In vitro and in vivo studies, we showed that the biphasic NF-kappaB activity is induced both in hemoglobin-induced neuron injury and rabbit SAH model, and the early peak indicated injury role on neurons survive, and the late peak denied deteriorated role on neuron fate.

    Patient Care: If we can explore the exactly inflammatory mechanism after SAH, and get a thorough description of NF-kappaB activation, we will get a more efficient anti-inflammation way to treat brain injury induced by SAH.

    Learning Objectives: We aim to discover the molecular mechanism of brain injury after SAH, especially the NF-kappaB activation in vivo and vitro experiment.

    References:

We use cookies to improve the performance of our site, to analyze the traffic to our site, and to personalize your experience of the site. You can control cookies through your browser settings. Please find more information on the cookies used on our site. Privacy Policy