Biomedical subjects

Nobuo Nagai

Publications and source records attributed to Nobuo Nagai.

10 recordsLinked to original sources

PubMed2006

Delayed perfusion phenomenon in a rat stroke model at 1.5 T MR: an imaging sign parallel to spontaneous reperfusion and ischemic penumbra?

INTRODUCTION: Delayed perfusion (DP) sign at MR imaging was reported in stroke patients. We sought to experimentally elucidate its relation to spontaneous reperfusion and ischemic penumbra. METHODS: Stroke was induced by photothrombotic occlusion of middle cerebral artery in eight rats and studied up to 72 h using a 1.5 T MR scanner with T2 weighted imaging (T2WI), diffusion weighted imaging (DWI), and dynamic susceptibility contrast-enhanced perfusion weighted imaging (DSC-PWI). Relative signal intensity (rSI), relative lesion volume (rLV), relative cerebral blood flow (rCBF), PWI(rLV)-DWI(rLV) mismatch (penumbra) and DP(rLV) were quantified and correlated with neurological deficit score (NDS), triphenyl tetrazolium chloride (TTC) staining, microangiography (MA) and histopathology. RESULTS: The rSI and rLV characterized this stroke model on different MRI sequences and time points. DSC-PWI reproduced cortical DP in all rats, where rCBF evolved from 88.9% at 1 h through 64.9% at 6 h to 136.3% at 72 h. The PWI(rLV)-DWI(rLV) mismatch reached 10+/-5.4% at 1 h, remained positive through 12 h and decreased to -3.3+/-4.5% at 72 h. The incidence and rLV of the DP were well correlated with those of the penumbra (p<0.01, r(2)=0.85 and p<0.0001, r(2)=0.96, respectively). Shorter DP durations and more collateral arterioles occurred in rats without (n=4) than with (n=4) cortex involvement (p<0.05). Rats without cortex involvement tended to earlier reperfusion and a lower NDS. Microscopy confirmed MRI, MA and TTC findings. CONCLUSIONS: In this rat stroke model, we reproduced clinically observed DP on DSC-PWI, confirmed spontaneous reperfusion, and identified the penumbra extending to 12h post-ischemia, which appeared interrelated.

Animals↗

PubMed2005

Role of tissue plasminogen activator/plasmin cascade in delayed neuronal death after transient forebrain ischemia.

We studied the possible involvement of the tissue plasminogen activator (t-PA)/plasmin system on both delayed neuronal death in the hippocampus and the associated enhancement of locomotor activity in rats, after transient forebrain ischemia induced by a four-vessel occlusion (FVO). Seven days after FVO, locomotor activity was abnormally increased and, after 10 days, pyramidal cells were degraded in the CA1 region of the hippocampus. FVO increased the t-PA antigen level and its activity in the hippocampus, which peaked at 4 h. Both the enhanced locomotor activity and the degradation of pyramidal cells were significantly suppressed by intracerebroventricular injection of aprotinin, a plasmin inhibitor, at 4 h but not during FVO. These results suggest the importance of the t-PA/plasmin cascade during the early pathological stages of delayed neuronal death in the hippocampus following transient forebrain ischemia.

Animals↗

PubMed2005

Dynamic susceptibility contrast-enhanced perfusion MR imaging at 1.5 T predicts final infarct size in a rat stroke model.

The purpose of the present animal experiment was to determine whether source images from dynamic susceptibility contrast-enhanced perfusion weighted imaging (DSC-PWI) at a 1.5T MR scanner, performed early after photochemically induced thrombosis (PIT) of cerebral middle artery (MCA), is feasible to predict final cerebral infarct size in a rat stroke model. Fifteen rats were subjected to PIT of proximal MCA. T2 weighted imaging (T2WI), diffusion-weighted imaging (DWI), and contrast-enhanced PWI were obtained at 1 h and 24 h after MCA occlusion. The relative lesion size (RLS) was defined as lesion volume/brain volume x 100% and measured for MR images, and compared with the final RLS on the gold standard triphenyl tetrazolium chloride (TTC) staining at 24 h. One hour after MCA occlusion, the RLS with DSC-PWI was 24.9 +/- 6.3%, which was significantly larger than 17.6 +/- 4.8% with DWI (P < 0.01). At 24 h, the final RLS on TTC was 24.3 +/- 4.8%, which was comparable to 25.1 +/- 3.5%, 24.6 +/- 3.6% and 27.9 +/- 6.8% with T2WI, DWI and DSC-PWI respectively (P > 0.05). The fact that at 1 h after MCA occlusion only the displayed perfusion deficit was similar to the final infarct size on TTC (P > 0.05) suggests that early source images from DSC-PWI at 1.5T MR scanner is feasible to noninvasively predict the final infarct size in rat models of stroke.

Animals↗

PubMed2004

Rat cerebral ischemia induced with photochemical occlusion of proximal middle cerebral artery: a stroke model for MR imaging research.

A stroke model in rats with photochemically induced thrombosis (PIT) of proximal cerebral middle artery (MCA) is introduced for magnetic resonance imaging (MRI) study. Thirty-seven rats subjected to surgical and optical procedures for inducing the PIT models were scanned using a 1.5-T scanner with T1-weighted imaging (T1WI), T2-weighted imaging (T2WI), diffusion-weighted imaging (DWI), and contrast-enhanced perfusion-weighted imaging (PWI) at 1 h and 24 h after MCA occlusion. The penumbra evolution and PWI-derived parameters including relative cerebral blood volume (rCBV) and relative cerebral blood flow (rCBF) were monitored; and the relative lesion size (RLS) was compared with the final RLS on the gold standard triphenyl tetrazolium chloride (TTC) staining at 24 h. The results showed that the focal cerebral ischemic lesions were detectable in all rats with different MR approaches. The lesion on PWI at 1 h and on all MR images at 24 h was matched well with that seen on TTC staining; the peri-infarct area decreased from 6.2 +/- 7.2% of the brain volume at 1 h to 0.3 +/- 5.6% at 24 h. Compared to that in the contralateral hemisphere, rCBV in ischemic region was 52.6 +/- 21.4 and 40.0 +/- 15.8% (p > 0.05), and rCBF was 64.6 +/- 11.2 and 47.3 +/- 11.1% (p < 0.05) at 1 h and 24 h respectively. The present PIT model in rats has been successfully adopted for MRI research, which might be feasible for certain stroke studies and should be beneficial for the evaluation on effects of potential diagnostic and therapeutic approaches.

Animals↗

PubMed2004

Visualization of stroke with clinical MR imagers in rats: a feasibility study.

This experiment was conducted in compliance with the guidelines of the International Committee on Thrombosis and Hemostasis and the current institutional regulations for use and care of laboratory animals. The purpose of the present study was to report the feasibility of using clinical magnetic resonance (MR) imaging devices for depiction of stroke in a rat model. Twenty-four rats with photochemically induced thrombosis of the middle cerebral artery were examined at superacute (1 hour, n = 24), acute (12 hours, n = 12), and subacute (24 hours, n = 12) phases with 1.5-T MR imaging weighted for T1, T2, diffusion, and gadopentetate dimeglumine-enhanced perfusion. With reasonable signal-to-noise ratio and imaging times, ischemic lesions were well distinguished on MR images as validated qualitatively and quantitatively with postmortem standard-of-reference techniques, including volume-rendered computed tomography, microangiography, and histochemistry. In the superacute phase, the perfusion defect at perfusion-weighted MR imaging was well matched with microangiographic and pathologic findings (P > .05). There was no difference in lesion size at perfusion-weighted MR imaging between superacute and subacute phases (P > .05). Performance of certain stroke-related research in rats is feasible with clinical MR imagers.

Animals↗

PubMed2004

Microplasmin reduces ischemic brain damage and improves neurological function in a rat stroke model monitored with MRI.

BACKGROUND AND PURPOSE: Microplasmin (microPli), a derivative of plasmin lacking the 5 "kringle" domains, was studied in a rat thrombotic stroke model with MRI monitoring. METHODS: Brain ischemia was induced by middle cerebral artery (MCA) occlusion with photochemically induced thrombosis. Brain tissue damage was assessed at 1 hour and 24 hours after MCA occlusion by MRI and 2,3,5-triphenyl tetrazolium chloride (TTC) staining. Neurological symptoms were scored at 24 hours. Animals with insufficient evidence of significant jeopardized brain tissue on perfusion-weighted imaging (PWI) at 1 hour were excluded before randomization. Included animals were randomized (blinded) to controls (solvent), 7.5 or 10 mg/kg microPli, administered as an intravenous bolus 90 minutes after MCA occlusion (n=8 per dose group). RESULTS: microPli treatment reduced cerebral damage measured by TTC staining at 24 hours, from 250+/-69 mm3 (mean+/-SD) in controls to 150+/-30 and 170+/-62 mm3 with 7.5 and 10 mg/kg microPli, respectively; it reduced the expansion of the PWI positive area between 1 and 24 hours, and it reduced neurological deficits from a Bederson score of 7 (6 to 9) in controls to 4.5 (3 to 8) and 4 (3 to 6), with 7.5 and 10 mg/kg microPli, respectively (median and rangeP<0.05 for each dose versus controls for all parameters). CONCLUSIONS: Bolus intravenous microPli given 90 minutes after thrombotic MCA occlusion in rats reduces cerebral ischemic damage and improves neurological dysfunction, suggesting that microPli could be beneficial in ischemic stroke patients.

Animals↗

PubMed2003

Essential role of endogenous tissue plasminogen activator through matrix metalloproteinase 9 induction and expression on heparin-produced cerebral hemorrhage after cerebral ischemia in mice.

Cerebral hemorrhage associated with antithrombotic and thrombolytic therapy in acute stroke continues to present a major clinical problem. Rupture of the cerebral microvasculature involves the degradation and remodeling of extracellular matrix. Here we demonstrated that the delayed administration of heparin 3 hours after photothrombotic middle cerebral artery occlusion (MCAO) caused cerebral hemorrhage in wild-type (WT) mice but not in tissue plasminogen activator (tPA)-deficient knockout (KO) mice. Heparin administration increased tPA activity and its mRNA expression at 6 and 12 hours after MCAO in the ischemic hemispheres of WT mice. The expression of tPA was enhanced in microglial cells in the ischemic border zone. We also observed an exacerbation of matrix metalloproteinase (MMP) 9 expression at the mRNA level and its conversion to an active form after heparin administration in the ischemic hemisphere in WT mice but not in tPA KO mice. The increased MMP 9 expression was localized in microglial cells and endothelial cells. These findings suggest that endogenous tPA, through the enhancement of MMP 9 expression and proteolytic activation, plays an essential role in the pathogenesis of heparin-produced cerebral hemorrhage. Targeting tPA, MMP 9, or both may provide a new approach for preventing cerebral hemorrhage associated with antithrombotic therapy for stroke in humans.

Animals↗

PubMed2003

Survivin-dependent angiogenesis in ischemic brain: molecular mechanisms of hypoxia-induced up-regulation.

Approaches to regulating angiogenesis in the brain, which may diminish parenchymal damage after stroke, are lacking. Survivin, the inhibitor of apoptosis protein, is up-regulated in vitro in vascular endothelial cells by angiogenic factors, including vascular endothelial cell growth factor (VEGF). To evaluate the in vivo role of survivin in the brain in response to hypoxia/ischemia, we used a mouse model of stroke and show that 2 days after permanent middle cerebral artery occlusion, survivin is uniquely expressed by microvessels that form in the peri-infarct and infarct regions. The extent of vascularization of the infarct is dependent on expression of survivin, since vessel density is significantly reduced in mice with heterozygous deficiency of the survivin gene (survivin+/- mice), even though infarct sizes were not different. Hypoxia alone induces survivin expression in the brain, by cultured endothelial cells and by embryonic stem cells, but this response is at least partially independent of VEGF, hypoxia inducible factor 1alpha, or placental growth factor. Delineating the spatiotemporal pattern of expression of survivin after stroke, and the molecular mechanisms by which this is regulated, may provide novel approaches to therapeutically optimize angiogenesis in a variety of ischemic disorders.

Animals↗

PubMed2002

Permissive proteolytic activity for visual cortical plasticity.

The serine protease, tissue-type plasminogen activator (tPA) is a key regulator of extracellular proteolytic cascades. We demonstrate a requirement for tPA signaling in the experience-dependent plasticity of mouse visual cortex during the developmental critical period. Proteolytic activity by tPA in the binocular zone was typically increased within 2 days of monocular deprivation (MD). This regulation failed to occur in glutamic acid decarboxylase (GAD) 65 knockout mice, an animal model of impaired ocular dominance plasticity because of reduced gamma-aminobutyric acid (GABA)-mediated transmission described previously. Loss of responsiveness to the deprived eye consequent to MD was conversely suppressed in mice lacking tPA despite normal levels of neuronal activity. Plasticity was restored in a gene dose-dependent manner, or by direct tPA infusion. Permissive amounts of tPA may, thus, couple functional to structural changes downstream of the excitatory-inhibitory balance that triggers visual cortical plasticity. Our results not only support a molecular cascade leading to neurite outgrowth after sensory deprivation, but also identify a valuable tool for further proteomic and genomic dissection of experience-dependent plasticity downstream of electrical activity.

Animals↗

PubMed2002

Tissue-type plasminogen activator has paradoxical roles in focal cerebral ischemic injury by thrombotic middle cerebral artery occlusion with mild or severe photochemical damage in mice.

The role of endogenous tissue-type plasminogen activator (tPA) in focal cerebral ischemic injury (FCII) after middle cerebral artery occlusion was studied using tPA gene-deficient (KO) mice and their wild-type (WT) littermates. The middle cerebral artery was occluded by thrombi induced by three different intensities of photochemical damage, a method that was newly introduced in mice. In both WT and KO mice, the intensity-dependent increase of FCII size was observed. The FCII size in tPA WT mice was smaller than in KO mice in cases of mild damage, whereas the FCII size was larger in WT mice than in KO mice in cases of severe damage. There was no difference in FCII size between WT and KO mice in cases of moderate damage. The number of microthrombi also increased with damage intensity in both WT and KO mice, but was less in WT mice at all intensities of damage. The results support the validity of the model of thrombotic occlusion by photochemical damage in mice, and suggest that endogenous tPA protects FCII through thrombolytic action on transient occlusion of middle cerebral artery with mild damage, but deteriorates on persistent occlusion with severe damage.

Animals↗