Jian-Xin Liu

Effect of ischemic postconditioning on cerebral edema and the AQP4 expression following hypoxic-eschemic brain damage in neonatal rats

BackgroundA rat model for neonatal hypoxic-ischemic brain damage (HIBD) was established to observe the effect of ischemic postconditioning (IPostC) on cerebral edema and the AQP4 expression following HIBD and to verify the neuroprotection of IPostC and the relationship between changes of AQP4 expression and cerebral edema.MethodsWater content was measured with dry-wet method, and AQP4 transcription and the protein expression of the lesions were detected with real-time PCR and immunohistochemistry staining, respectively.ResultsWithin 6–48 hours, the degree of ipsilateral cerebral edema was significantly lower in IPostC-15 s/15 s group than in HIBD group. Similar to the HIBD group, the AQP4 transcription and expression in the IPostC group showed a downward and then upward trend. But the expression was still more evident in the HIBD group than in the IPostC-15 s/15 s group. From 24 to 48 hours, IPostC-15 s/15 s decreased the slowing down expression of AQP4.ConclusionIPostC has neuroprotective effect on neonatal rats with HIBD and it may relieve cerebral edema by regulating the expression of AQP4.

Newly generated neurons at 2 months post-status epilepticus are functionally integrated into neuronal circuitry in mouse hippocampus

Emerging evidence has linked chronic temporal lobe epilepsy to dramatically reduced neurogenesis in the dentate gyrus. However, the profile of different components of neurogenesis in the chronically epileptic hippocampus is still unclear, especially the incorporation of newly generated cells. To address the issue, newly generated cells in the sub-granular zone of the dentate gyrus were labeled by the proliferation marker bromodeoxyuridine (BrdU) or retroviral vector expressing green fluorescent protein 2 months after pilocarpine-induced status epilepticus. The newly generated neurons that extended axons to CA3 area or integrated into memory circuits were visualized by cholera toxin B subunit retrograde tracing, and detecting activation of BrdU(+) cells following a recall of spatial memory test at the chronic stage of TLE. We found that the microenvironment was still able to sustain significant neuronal differentiation of newly generated cells at 2 months post-status epilepticus time-point, and newly added neurons into granular cell layer were still able to integrate into neuronal circuitry, both anatomically and functionally. Quantified analyses of BrdU(+) or Ki-67(+) cells demonstrated that there was a reduced proliferation of progenitor cells and diminished survival of newly generated cells in the epileptic hippocampus. Both decreased levels of neurotrophic factors in the surrounding milieu and cell loss in the CA3 area might contribute the decreased production of new cells and their survival following chronic epilepsy. These results suggest that decreased neurogenesis in the chronically epileptic hippocampus 2 months post status epilepticus is not associated with altered integration of newly generated neurons, and that developing strategies to augment hippocampal neurogenesis in chronic epilepsy might be protective.

Reduced abnormal integration of adult-generated granule cells does not attenuate spontaneous recurrent seizures in mice

Epileptic seizures lead to aberrant hippocampal neurogenesis, including increased proliferation of neural progenitors and abnormal integrations of newly generated granule cells - hilar ectopic granule cells (EGCs), mossy fiber sprouting (MFS), and hilar basal dendrites (HBDs). Previous results from ablating hippocampal neurogenesis after acute seizures have been controversial with regards to the development of spontaneous recurrent seizures (SRSs). While ablation of hippocampal newborn cells was effective, a sufficient decrease of subsequent abnormal integrations in chronically epileptic hippocampus was not well-established in these studies. Evaluations of the role of aberrant neurogenesis in epileptogenesis were therefore inconclusive. In this study, we ablated the hippocampal neurogenesis by methylazoxymethanol acetate (MAM) treatment both before and after pilocarpine induced status epilepticus (SE). We found that an overall ablation of newborn granule cells and a protracted delay after the cell ablation are required to eliminate subsequent abnormal integrations, including EGCs, MSF and HBDs. However, there were no alterations in frequency, duration and severity of chronic seizures were demonstrated following this regime. The current findings provide novel evidences that an overall decrease of abnormal integrations via cell ablation cannot exert significant effects on the development of SRSs at least in the model used in this study.

Reduced expression of Phospholipase C beta in hippocampal interneuron during pilocarpine induced status epilepticus in mice

We investigated localization of Phospholipase C beta (PLCβ1 and PLCβ4) in laminaes of dorsal hippocampus and different subtypes of hippocampal interneurons in normal Kunming mouse, and their progressive changes during pilocarpine induced status epilepticus (SE) by quantitative immunohistochemistry and real time PCR. PLCβ1 was observed in the pyramidal layer of CA1-3 area, hilus of the dentate gyrus, whereas PLCβ4 was mainly expressed in calcium binding protein positive interneurons, i.e. calbindin, calretinin, parvalbumin positive interneurons in the strata oriens, radiatum of the CA area and hilus of the dentate gyrus. During pilocarpine induced SE, a temporary down-regulation of PLCβ4 in the interneurons of CA area at SE 30min, and a progressive reduction of PLCβ1/PLCβ4 in dentate hilar cells were demonstrated. These findings confirm and extend the regional specific distribution of PLCβ1 and PLCβ4 immunoreactivity in mouse hippocampus, and suggest that PLCβ1 and PLCβ4 may play an important role in maintenance of the status epilepticus.

Ablation of aberrant neurogenesis fails to attenuate cognitive deficit of chronically epileptic mice

Pilocarpine-induced acute seizures strongly induce aberrant hippocampal neurogenesis, characterized by increased proliferation of neural progenitors and abnormal integrations of newly generated granule cells - hilar ectopic granule cells (EGCs), mossy fibre sprouting (MFS), and hilar basal dendrites (HBDs), which may disturb hippocampal neuronal circuits and thus contribute to cognitive impairment in temporal lobe epilepsy (TLE) patients and animal models. Previous studies via ablating hippocampal neurogenesis after acute seizures produced inconsistent results regarding the development of long-term cognitive impairment. Furthermore, a sufficient decrease of subsequent abnormal integrations in chronically epileptic hippocampus was not well-established in these studies. Therefore, the link between seizure-induced aberrant hippocampal neurogenesis and cognitive decline associated with epilepsy is still in need to be clarified. In this study, the mice were injected with methylazoxymethanol acetate (MAM) both before and after pilocarpine-induced status epilepticus (SE) to achieve an overall ablation of newborn cells contributing to the pathological recruitment. In addition, a protracted time point was chosen for behavioral testing considering it takes a fairly long time for newborn granule cells to adequately develop abnormal integrations, especially MFS. Although an overall reduction of abnormal integrations, including EGCs, MFS and HBDs was confirmed following the ablation regime, the performance of ablated and non-ablated mice in the Morris Water Maze (MWM) task did not differ. The current findings therefore provide novel evidences that ablation of neurogenesis with an overall decrease of abnormal integrations cannot attenuate subsequent cognitive impairment at least in the model used in this study.

Aspirin attenuates spontaneous recurrent seizures in the chronically epileptic mice

Abstract Objective: Neuroinflammatory processes are pathologic hallmarks of both experimental and human epilepsy, and could be implicated in the neuronal hyperexcitability. Aspirin represents one of the non-selective nonsteroidal anti-inflammatory drugs with fewer side effects in long-term application. This study was carried out to assess the anti-epileptic effects of aspirin when administered during the chronic stage of temporal lobe epilepsy [TLE] in mice. The alteration of hippocampal neurogenesis was also examined for raising a possible mechanism underlying the protective effect of anti-inflammatory treatment in the TLE. Methods: Two months after pilocarpine-induced status epilepticus, the chronically epileptic mice were treated with aspirin (20 mg, 60 mg or 80 mg/kg) once a day for 10 weeks. Spontaneous recurrent seizures were monitored by video camera for 2 weeks. To evaluate the profile of hippocampal neurogenesis, the newly generated cells in the dentate gyrus were labeled by the proliferation marker BrdU. The newborn neurons that extended axons to CA3 area were visualized by cholera toxin B subunit retrograde tracing. Results: Administration of aspirin with a dosage of 60 mg or 80 mg/kg initiated at 2 months after pilocarpine-induced status epilepticus significantly reduced the frequency and duration of spontaneous recurrent seizures. Aspirin treatment also increased the number of newborn neurons with anatomic integration through improving the survival of the newly generated cells. Conclusion: Aspirin treatment during the chronic stage of TLE could attenuate the spontaneous recurrent seizures in mice. Promotion of hippocampal neurogenesis and inhibition of COX-PGE2 pathway might partly contribute to this anti-epileptic effect. Highlights• Aspirin attenuates spontaneous recurrent seizures of chronically epileptic mice• Aspirin increases neurogenesis of chronically epileptic hippocampus by improving the survival of newly generated cells• Promotion of hippocampal neurogenesis and inhibition of COX-PGE2 pathway might partly contribute to anti-epileptic effects of aspirin.

MTEP impedes the neuronal polarization and the activity of the Akt–NF‐κB pathway in rat hippocampal neurons

Metabotropic glutamate receptor 5 (mGluR5) is extensively involved in neural survival, differentiation, dendritic morphology, synaptic plasticity, and neural circuit formation. However, little is known about its role in neuronal polarization and axon outgrowth. In this study, we applied the selective agonist (RS)‐2‐chloro‐5‐hydroxyphenylglycine sodium salt and antagonist 3‐[(2‐methyl‐4‐thaizolyl) ethynyl] pyridine (MTEP) of mGluR5 to the cultured hippocampal neurons to observe the neuronal polarization and axon outgrowth, and further explored the possible intracellular signal transduction pathway. The results demonstrated that MTEP administration significantly attenuates the proportion of polarized neurons and the length of the axon, indicated by SMI312 (an axon marker) and Tuj‐1 (a marker of all the neurites) double‐labeling immunofluorescence. Western blot analysis showed that MTEP administration also inhibited the activation of AKT and nuclear translocation of nuclear factor‐κB (NF‐κB) p65, and decreased the phosphorylation of p65 as well. Furthermore, Akt inhibitor LY294002 treatment resulted in neuronal polarization delay and axon outgrowth retardation, while suppressing the phosphorylation and nuclear translocation of p65. We concluded that mGluR5 could regulate neuronal polarity and axon outgrowth during the morphological differentiation of rat developing neurons, and the intracellular signaling pathway of Akt–NF‐κB might be involved in the action of mGluR5.

Systematic review and meta-analysis of the efficacy of different exercise programs in pilocarpine induced status epilepticus models

PURPOSE To conduct a systematic review and meta-analysis of studies testing exercise in animal models of pilocarpine induced status epilepticus (SE) and to compare the efficacy of different training strategies used in those studies. METHODS We searched 2 online databases (Pubmed and Web of Science) for studies analyzing the efficacy of different trainings in pilocarpine-induced SE models. Training was categorized into forced physical training (PT), voluntary PT and resistance PT. Two reviewers independently extracted data on study quality, behavioral seizures, and histological, chemical and cognitive outcomes. Data were pooled by means of a meta-analysis. RESULTS Among 17 selected studies; 174 animals from 8 studies with 10 comparison groups showed that exercise intervention after induction of SE significantly decreased spontaneous recurrent seizures with [mean difference (MD)=-1.80, 95% confidence interval (CI): -3.22, -0.37, p=0.02] and 60 animals showed statistically significant decrease in latency in Morris water maze (standardized mean difference (SMD)=-2.57, 95% CI: -4.06, -1.08, p=0.0007). Although not statistically significant, still a remarkable increase in number of CA1 neurons and hippocampal BDNF level (MD=2.27, [95% CI: -1.20, 5.73], p=0.19, SMD=1.07, [95% CI: -0.36, 2.51], p=0.14 respectively) and a decrease in mossy fibers sprouting (SMD=-1.03, [95% CI: -3.06, 1.00], p=0.32) were observed. PT interventions in 72 animals before induction of SE showed favorable increase in latency to develop SE (MD=8.34, [95% CI: -3.10, 19.78], p=0.15) but no remarkable improvements in latency for the first motor sign and motor signs intensity. CONCLUSIONS PT after SE reduces the recurrent seizures and improves the morphological, biochemical and cognitive profiles of pilocarpine epileptic models. Resistance PT was identified as particularly effective in reducing behavioral seizures. The efficacy of training was also dependent upon duration.