Research reportTAT-PEP, a novel blocker of PirB, enhances the recovery of cognitive function in mice after transient global cerebral ischemia
Introduction
Global cerebral ischemia, which is usually seen in clinical conditions like cardiac arrest, drowning or systemic hypotension during surgery [1], is associated with high mortality rates [2]. Global cerebral ischemia mainly attack the vulnerable brain region like the CA1 region in hippocampus [3], leading to neurological deficit in memory [4] and executive function [5]. The fibrinolytic therapy and anti-platelet treatment are only effective for acute brain ischemia, but not able to avoid the cognitive dysfunctions and long-term neurological deficit resulting from ischemia.
Loss of neurons and axonal regeneration difficulty after ischemia in the hippocampus are the main reasons for cognitive dysfunctions [6]. Nogo-A, myelin-associated glycoprotein (MAG), and oligodendrocyte-myelin glycoprotein (OMgp) are known as three myelin proteins that limit axonal regeneration in the central nerve system (CNS) [7]. Paired immunoglobulin like receptor B (PirB) is also a receptor of the three myelin proteins. The knockout of PirB or NgR1 could promote neurite outgrowth in CNS [8], [9]. In an Alzheimer’s Model, it was reported that PirB acted as receptors for oligomeric forms of Aβ42, which may participate in synaptic loss and cognitive impairment in AD progression [10]. Another study showed that the numbers of midline crossing fibers from the undamaged corticospinal tract of PirB knockout mouse were increased after focal cerebral ischemia model. Function recovery of PirB knockout mouse was enhanced after injury [8]. In addition, Anti-PirB treatment of NgR−/− neurons completely restored the neurite outgrowth on myelin [11]. It appears that PirB is more important than NgR in neurite outgrowth. Our previous study also showed that PirB was significantly increased in neurons after brain ischemia injury [12]. With all these findings, inhibiting the activities of PirB until Nogo-A is expressed to peak seems to be an effective means to promote the long-term outcome of brain ischemia injury. This evidence suggests that PirB function crucially contributes to the exacerbation of brain damage and the restriction of functional recovery by limiting plasticity during cerebral ischemia-reperfusion injury. It is plausible to hypothesize that PirB is a novel potential therapeutic target for stroke and that inhibiting PirB function exerts a more powerful effect than deleting only a subset of its ligands.
Protein transduction domains (protein transducer domain, PTDs), a novel structure for carrying various macromolecules through the blood-brain barrier, have been receiving great attention in the recent years. In this study, we fused human immunodeficiency virus (HIV) transactivator of transcription (TAT), a widely used PTD, with PirB extracellular region (PEP) to generate the TAT-PEP fusion protein. The generation of TAT-PEP was shown in the supplementary material.
Thus, the main purpose of this study is to explore the effect of TAT-PEP on cognitive function as well as motor functional recovery after global cerebral ischemia. This work investigated whether TAT-PEP more effectively promoted neuritis outgrowth and enhanced functional recovery in mice that underwent bilateral common carotid artery occlusion (BCCAO).
Section snippets
Oxygen-glucose deprivation (OGD) model
To test the neuroprotective function of TAT-PEP against ischemic injury in vitro, the OGD model was used in our study as previously described [13]. In brief, we designed several groups: Control (Normal cells), OGD, OGD + TAT-PEP (150 μg/L). HT22 cells were cultured in neurobasal medium without glucose (Invitrogen, Carlsbad, CA, USA) in a humidified incubator filled with an anoxic gas mixture (5% CO2 and 95% N2) at 37 °C for 1 h and were then cultured in normal medium in the presence or absence of
TAT-PEP increased cell viability and promoted neurite outgrowth in HT22 cells exposed to OGD
OGD significantly decreased the viability of HT22 cells. Treatment of cells with TAT-PEP at a concentration of 150 μg/L markedly increased the viability of HT22 cells subjected to OGD (Fig. 1A). We also found that TAT-PEP significantly promoted the growth of the neurite of HT22 cells subjected to OGD. It seems that TAT-PEP completely recover the growth of neurite of HT22 cells since there was no significant difference between the BCCAO group and the Con group (Fig. 1B, C).
TAT-PEP effectively transduced into the brain and alleviated hippocampal injury after transient global cerebral ischemia
The expression of PirB
Discussion
Global cerebral ischemia is commonly seen in clinical conditions such as cardiac arrest and leads to neuronal damage including selective neuronal loss in the hippocampus [20], which is involved in memory processing [21]. Therefore, memory impairment is the most common neurological dysfunction seen after ischemic injury. In addition to delayed neurodegeneration (ahead of apoptosis) and apoptosis all participated at the time of injury [22]. There is currently no effective therapy.
Nogo-A, MAG, and
Conclusion
The present results showed that an intraperitoneal injection of TAT-PEP could effectively deliver TAT-PEP to the brain through BBB. In addition, TAT-PEP reduced neuronal apoptosis and degeneration, and increased neuronal survival in the hippocampus. The promotion of neurite regeneration and neuron survival after TAT-PEP treatment could account for the long-term functional recovery post BCCAO. This method could potentially provide a viable therapy for improving the prognosis of cerebrovascular
Formatting of funding sources
This work was supported by the National Natural Science Foundation of China (Grant NO. 81473488, 81471373, 81501207), the Overseas, Hong Kong & Macao Scholars Collaborated Researching Fund (Grant 81529004), the National Key Technology Research and Development Program of the Ministry of Science and Technology of China (2012BAI20B02).
Acknowledgments
We thank the State Key Laboratory of Military Stomatology (Department of Anesthesiology, School of Stomatology, The Fourth Military Medical University, Xi’an, China.) for providing the TAT-PEP.
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These authors contributed equally to this work.