Jinsheng Zeng

Beclin 1 knockdown inhibits autophagic activation and prevents the secondary neurodegenerative damage in the ipsilateral thalamus following focal cerebral infarction.

Cerebral infarction can cause secondary degeneration of thalamus and delay functional recovery. However, the mechanisms underlying secondary degeneration are unclear. The present study aimed to determine the occurrence and contribution of autophagy to the thalamic degeneration after cerebral infarction. Focal cerebral infarction was induced by distal middle cerebral artery occlusion (MCAO). Autophagic activation, Beclin 1 expression and amyloid β (Aβ) deposits were determined by immunofluorescence, immunoblot and electron microscopy. Secondary damage to thalamus was assessed with Nissl staining and immunofluorescence analysis. Apoptosis was determined using TUNEL staining. The contribution of autophagy to the secondary damage was evaluated by shRNA-mediated downregulation of Beclin 1 and the autophagic inhibitor, 3-methyladenine (3-MA). The potential role of Aβ in autophagic activation was determined with N-[N-(3, 5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT). The results showed that the conversion of LC3-II, the formation of autophagosomes, and the levels of activated cathepsin B and Beclin 1 were significantly increased in the ipsilateral thalamus at 7 and 14 days after MCAO (p < 0.05 or 0.01). Both Beclin 1 knockdown and 3-MA treatment significantly reduced LC3-II conversion and autophagosome formation, which were accompanied by obvious decreases in neuronal loss, gliosis and apoptosis in the ipsilateral thalamus (p < 0.05 or 0.01). Additionally, DAPT treatment markedly reduced Aβ deposits, which coincided with decreases in LC3-II conversion and autophagosome formation (p < 0.01). These results suggest that inhibition of autophagy by Beclin 1 knockdown can attenuate the secondary thalamic damage after focal cerebral infarction. Furthermore, Aβ deposits may be involved in the activation of autophagy.

A prospective study of secondary degeneration following subcortical infarction using diffusion tensor imaging

Background: Secondary degeneration of the pyramidal tract distal to the primary lesion after a stroke has been detected by some studies using diffusion tensor imaging (DTI) but its potential clinical significance and the degeneration of the fibre tract proximal to the primary lesion have received little attention. Methods: Twelve patients underwent DTI on the 1st, 4th and 12th week following a subcortical infarct involving the posterior limb of the internal capsule, and 12 age and sex matched controls underwent DTI once. The DTI parameters mean diffusivity and fractional anisotropy (FA), and the clinical scores before DTI examination, including the National Institutes of Health Stroke Scale (NIHSS), the Fugl–Meyer (FM) scale and the Barthel index (BI), were assessed. The relations between the per cent changes in DTI parameters and clinical scores were analysed. Results: From the 1st to the 12th week after stroke onset, FA values decreased (p<0.01, respectively) in the fibre tract above and below the internal capsule, and the NIHSS decreased (p<0.01) but the FM scale and BI increased (p<0.01, respectively) progressively. The per cent reductions in FA value in the fibre tract above and below the internal capsule were negatively correlated with the per cent changes in NIHSS and FM scale (p<0.05, respectively). Conclusions: Secondary degeneration of the fibre tract proximal and distal to a primary lesion can be detected by DTI clearly and quantitatively and deteriorates with time progressively, which may hamper functional recovery after a subcortical cerebral infarct.

Secondary Neurodegeneration in Remote Regions After Focal Cerebral Infarction: A New Target for Stroke Management?

Cerebral infarction-induced cessation of function in areas of the brain remote from, but connected to, the primary site of damage was termed “diaschisis” by von Monakow.1 Initially, the concept of diaschisis did not include morphological changes. However, accumulating evidence has shown that histopathologic changes also occur in nonischemic remote brain regions that have synaptic connections with the primary lesion site. For example, after cerebral infarction in the middle cerebral artery (MCA) territory, neuronal death, gliosis, and axonal degeneration have been found in the ipsilateral thalamus, substantia nigra (SN), and distal pyramidal tract, all of which lie outside the MCA territory.2–7 This kind of secondary neurodegeneration occurs selectively in such areas several days or weeks after stroke onset, and this can be detected by neuroimaging techniques.8 For quite a long time, the role of secondary degeneration in stroke recovery has not been well understood. Recently, emerging studies suggest that secondary degeneration is associated with neurological deficits and can predict motor outcome after stroke.9–15 In this review, we aimed to summarize the pathological and neuroimaging evidence of secondary neurodegeneration in the ipsilateral thalamus, SN, and pyramidal tract after MCA infarction and described its potential significance for stroke management. We searched PubMed from 1980 to September 2011, using the terms “cerebral infarction,” “middle cerebral artery,” “Wallerian degeneration,” “anterograde degeneration,” “retrograde degeneration,” and “transneuronal degeneration.” Further articles were identified from the references cited in those articles and through searches of our personal files. The final list of references was selected on the basis of originality and relevance to the topics covered in this review. ### Postmortem Studies Data from postmortem studies provide direct evidence of secondary neurodegeneration after focal cerebral infarction (Figure 1). Histopathologic examination at 4 months after MCA infarction revealed a delayed and selective decrease in the …

New-Onset Constipation at Acute Stage After First Stroke: Incidence, Risk Factors, and Impact on the Stroke Outcome

Background and Purpose— The prevalence of constipation after stroke varies from 30% to 60%. The incidence of new-onset constipation during the early stage of stroke remains uncertain. The present study was designed to investigate the prevalence of new-onset constipation, its risk factors, and its impact on stroke outcome in patients with their first stroke at acute stage. Methods— This is a prospective cohort study of 154 patients admitted with their first stroke. New-onset constipation during the first 4 weeks of stroke was recorded, using the Rome II criteria for constipation. Demographics, characteristics of the stroke, laboratory parameters, and use of medications were evaluated as risk factors for constipation. Death, recurrent stroke, and handicap at 12 weeks were regarded as poor outcome. The impact of constipation on poor outcome was also studied. Results— The cumulative incidence of new-onset constipation was 55.2% at 4 weeks poststroke. The occurrence of constipation was associated with dependence (P<0.01) and use of bedpan for defecation (P<0.05). Among patients with moderate stroke severity (NIHSS 4 to11) at baseline, constipation at 4 weeks was associated with a poor outcome at 12 weeks. Conclusions— New-onset constipation is a common complication of acute stroke. Its occurrence is associated with dependence and use of bedpan for defecation. Its development may predict a poor outcome at 12 weeks in patients with moderately severe stroke.

Nogo-A is involved in secondary axonal degeneration of thalamus in hypertensive rats with focal cortical infarction

We investigate whether Nogo-A is involved in the secondary axonal degeneration in the thalamus after distal middle cerebral artery occlusion (MCAO) in stroke-prone renovascular hypertensive rats (RHRSP). The expression of Nogo-A in ipsilateral ventroposterior nucleus (VPN) of the thalamus in RHRSP was observed at 1, 2 and 4 weeks after distal MCAO. In addition, intracerebroventricular infusion of NEP1-40, a Nogo-66 receptor (NgR) antagonist peptide, was administered starting 24 h after MCAO and continued for 1, 2 and 4 weeks, respectively. Axonal damage and regeneration were evaluated by analysis of the immunoreactivity (IR) of amyloid betaA4 precursor protein (APP), growth associated protein 43 (GAP-43) and microtubule associated protein 2 (MAP-2) in ipsilateral VPN of the thalamus at 1, 2 and 4 weeks after distal MCAO. Following ischemia, the expression of Nogo-A in oligodendrocytes increased persistently and its localization became redistributed around damaged axons and dendrites. Administration of NEP1-40 downregulated the expression of Nogo-A, reduced axonal injury and enhanced axonal regeneration. Our data suggest that Nogo-A is involved in secondary axonal degeneration and that inhibition of Nogo-A can reduce neuronal damage in the thalamus after distal MCAO.

Longitudinal Investigations on the Anterograde and Retrograde Degeneration in the Pyramidal Tract following Pontine Infarction with Diffusion Tensor Imaging

Background: Secondary degeneration following supratentorial stroke has been detected by some studies using diffusion tensor imaging (DTI), but the anterograde and retrograde degeneration in pyramidal tract after pontine infarction and its potential clinical significance are not well understood. Methods: Fourteen patients with a recent pontine infarct underwent three DTIs at week 1, week 4, and week 12 after onset. Fourteen age- and gender-matched controls underwent DTI once. Mean diffusivity and fractional anisotropy (FA) were measured. Neurological deficit, motor deficit and life independence were assessed with the NIH Stroke Scale, Fugl-Meyer scale and Barthel index, respectively, 2 h before each DTI examination. Results: FA values at the ipsilateral medulla and the proximal portion of the pyramidal tract, including centrum semiovale, internal capsule and cerebral peduncle, significantly decreased progressively from week 1 to week 12 (p < 0.01). The NIH Stroke Scale decreased; Fugl-Meyer scale and Barthel index increased significantly over the time points (p < 0.01). The absolute values of percent reduction of FA value at the ipsilateral medulla and the proximal portion of pyramidal tract correlated negatively with the absolute values of percent reduction of the NIH Stroke Scale and percent increase of the Fugl-Meyer scale. Conclusions: Progressive anterograde and retrograde degeneration in pyramidal tract revealed by DTI may hinder the process of neurological recovery after a pontine infarct. To confirm the clinical significance, future studies with a longer observation period and a larger sample size of patients with more homogeneous pontine infarcts are still needed.

Reduction of β-Amyloid Deposits by γ-Secretase Inhibitor is Associated with the Attenuation of Secondary Damage in the Ipsilateral Thalamus and Sensory Functional Improvement after Focal Cortical Infarction in Hypertensive Rats

Abnormal β-amyloid (Aβ) deposits in the thalamus have been reported after cerebral cortical infarction. In this study, we investigated the association of Aβ deposits, with the secondary thalamic damage after focal cortical infarction in rats. Thirty-six stroke-prone renovascular hypertensive rats were subjected to distal middle cerebral artery occlusion (MCAO) and then randomly divided into MCAO, vehicle, and N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester (DAPT) groups and 12 sham-operated rats as control. The DAPT was administered orally at 72 hours after MCAO. Seven days after MCAO, sensory function, neuron loss, and glial activation and proliferation were evaluated using adhesive removal test, Nissl staining, and immunostaining, respectively. Thalamic Aβ accumulation was evaluated using immunostaining and enzyme-linked immunosorbent assay (ELISA). Compared with vehicle group, the ipsilateral thalamic Aβ, neuronal loss, glial activation and proliferation, and the mean time to remove the stimulus from right forepaw significantly decreased in DAPT group. The mean time to remove the stimulus from the right forepaw and thalamic Aβ burden were both negatively correlated with the number of thalamic neurons. These findings suggest that Aβ deposits are associated with the secondary thalamic damage. Reduction of thalamic Aβ by γ-secretase inhibitor may attenuate the secondary damage and improve sensory function after cerebral cortical infarction.

Autophagosomes accumulation is associated with β-amyloid deposits and secondary damage in the thalamus after focal cortical infarction in hypertensive rats

J. Neurochem. (2012) 120, 564–573.

Neurogenesis and angiogenesis within the ipsilateral thalamus with secondary damage after focal cortical infarction in hypertensive rats

Neurogenesis and angiogenesis in the subventricular zone and peri-infarct region have been confirmed. However, newly formed neuronal cells and blood vessels that appear in the nonischemic ipsilateral ventroposterior nucleus (VPN) of the thalamus with secondary damage after stroke has not been previously studied. Twenty-four stroke-prone renovascular hypertensive rats were subjected to distal right middle cerebral artery occlusion (MCAO) or sham operation. 5′-Bromo-2′-deoxyuridine (BrdU) was used to label cell proliferation. Rats were killed at 7 or 14 days after the operation. Neuronal nuclei (NeuN), OX-42, BrdU, nestin, laminin+, BrdU+/nestin+, BrdU+/NeuN+, nestin+/GFAP+(glial fibrillary acidic protein), and BrdU+/laminin+ immunoreactive cells were detected within the ipsilateral VPN. The primary infarction was confined to the right somatosensory cortex. Within the ipsilateral VPN of the ischemic rats, the number of NeuN+ neurons decreased, the OX-42+ microglia cells were activated, and BrdU+ and nestin+ cells were detected at day 7 after MCAO and increased in number at day 14. Moreover, BrdU+/nestin+ cells and BrdU+/NeuN+ cells were detected at day 14 after MCAO. In addition, the ischemic rats showed a significant increase in vascular density in the ipsilateral VPN compared with the sham-operated rats. These results suggest that secondary damage with neurogenesis and angiogenesis of the ipsilateral VPN of the thalamus occurs after focal cortical infarction.

Marine compound Xyloketal B protects PC12 cells against OGD-induced cell damage.

Xyloketal B is a novel marine compound with unique chemical structure isolated from mangrove fungus Xylaria sp. (no. 2508). Recently, we have demonstrated that Xyloketal B is an antioxidant and can protect against oxidized low density lipoprotein (LDL)-induced cell injury. In the present study, we investigated whether Xyloketal B can protect against ischemia-induced cell injury in an in vitro oxygen glucose deprivation (OGD) model of ischemic stroke in PC12 cells. We found that Xyloketal B could directly scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical and protect PC12 cells against OGD insult. Furthermore, Xyloketal B alleviated OGD-induced mitochondria superoxide, mitochondria fragmentation and GTPase dynamin-related protein 1 (Drp1) overexpression as well as reduction of mitochondrial membrane potential. All together, the present study demonstrates that Xyloketal B protects PC12 cells against OGD-induced cell injury and that the anti-oxidative property and protective action on mitochondria may account for its neuroprotective actions.