The purpose of this study was to identify time-related changes in clinical, MRI, histopathologic, and immunohistochemical findings associated with ischemic stroke in dogs. Additionally, the association of cerebrospinal fluid (CSF) and tissue levels of interleukin (IL)-6 with clinical prognosis was assessed. Ischemic stroke was induced by permanent middle cerebral artery occlusion (MCAO) in nine healthy experimental dogs. The dogs were divided into three groups according to survival time and duration of the experimental period: group A (survived only 1 day), group B (1-week experimental period), and group C (2-week experimental period). Neurologic status was evaluated daily. Magnetic resonance imaging (MRI) was performed according to a predetermined schedule. Concentration of IL-6 in CSF was measured serially after ischemic stroke. Postmortem examination was performed for all experimental dogs. During histopathological examination, variable degrees of cavitation and necrosis due to neuronal cytopathic effects, such as pyknotic nuclei and cytoplasmic shrinkage, were observed on the affected side of the cerebral cortex in all dogs. Immunohistochemistry specific for IL-6 showed increased expression in the ischemic lesions. CSF IL-6 concentrations and ischemic lesion volumes 1 day after ischemic stroke were significantly higher in group A compared to groups B and C.
Animals ; Brain Ischemia/*etiology ; Dogs ; Female ; *Immunohistochemistry ; *Infarction, Middle Cerebral Artery ; *Magnetic Resonance Imaging ; Male ; Stroke/*pathology

This study aimed to investigate inflammatory edema after cerebral ischemia through 7.0T MRI and proton magnetic resonance spectroscopy (MRS). All SD rats were randomly divided into sham operated group and middle cerebral artery occlusion (MCAO)-1 day, -3 day and -7 day groups. MRI scan of the brain was performed on a 7.0 Tesla MRI scanner. The volume of positive signals in the ischemic side was detected by using a T2 weighted spinecho multislice sequence; the changes in the height of water-peak were measured with point resolved spectroscopy (PRESS) sequences; cortical edema was detected by using wet-dry weight method; the degrees of nerve injury were evaluated by Bederson neurological score system; double-labeling immunofluorescence technique was used to explore the molecular mechanisms of post-ischemia cerebral edema. The results showed that high T2WI signals were observed in MCAO-1 day, -3 day and -7 day groups, and the water-peak height and water-peak area of MCAO groups were higher than those of sham operated group (P<0.05). Neurological score results were consistent with the degree of brain edema, and a large number of microglia accumulated in the ischemic cortex. Our results suggested that non-invasive MRI technology with the advantage of high spatial resolution and tissue resolution can comprehensively and dynamically observe inflammatory edema after cerebral ischemia from a three-dimensional space, and contribute to evaluation and treatments in clinic.
Animals ; Brain ; diagnostic imaging ; pathology ; Brain Edema ; diagnostic imaging ; etiology ; Brain Ischemia ; complications ; CD11b Antigen ; metabolism ; Immunohistochemistry ; Infarction, Middle Cerebral Artery ; complications ; Inflammation ; diagnostic imaging ; etiology ; Magnetic Resonance Imaging ; methods ; Magnetic Resonance Spectroscopy ; Male ; Microglia ; metabolism ; Microscopy, Confocal ; Microscopy, Fluorescence ; Radiography ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Time Factors

OBJECTIVETo establish a general method of focal cerebral ischemia model in different varieties of mice.

METHODEach group of healthy adult KM and C57BL/6 mice were randomly divided into control group (n = 10) and MCAO group (n = 10). The mice in MCAO group were applied in the preparation of the MCAO model by intraluminal occlusion using monofilament. Twenty-four hours after operation,the neurologic function was evaluated,middle cerebral artery blood flow was monitored and the infarction volume was calculated by TTC staining, to evaluate the reliability of the model.

RESULTIn the MCAO group, the base value of the cerebral blood flow down of KM and C57BL/6 mice respectively was (81.65 ± 4.59)%, (83.68 ± 6.25)%. The neurological deficit score respectively was (2.30 ± 0.82), (2.50 ± 0.80). TTC staining can clearly show the infarction area, and relatively stable, 24 hours of the survival rate of KM and C57BL/6 mice were 100% and 80% respectively.

CONCLUSIONThe key link is the optimization and improvement of monofilament, temperature, anesthesia and so on. The modified intraluminal occlusion of MCAO using monofilament is a kind of reliable and simple method to establish experimental cerebral ischemia model in mice.

Animals ; Blood Flow Velocity ; Brain ; blood supply ; pathology ; physiopathology ; Brain Ischemia ; complications ; physiopathology ; Cerebrovascular Circulation ; Disease Models, Animal ; Infarction, Middle Cerebral Artery ; complications ; physiopathology ; Male ; Mice, Inbred C57BL ; Middle Cerebral Artery ; pathology ; physiopathology ; surgery ; Nervous System Diseases ; etiology ; physiopathology ; Species Specificity

This study is to observe the effect of ilexonin A (IA) on the expression of basic fibroblast growth factor (bFGF) and growth associated protein-43 (GAP-43), and neurogenesis after cerebral ischemia-reperfusion in rats and explore its possible mechanism of protecting neuronal injury. Models of middle cerebral artery occlusion (MCAO) were established in SD rats. Before and after two hours ischemia-reperfusion, IA (20 and 40 mg x kg(-1)) was injected immediately and on 3, 7, 14, and 28 d once a day. The neurological severity was evaluated by neurological severity scores (NSS); neuronal injury in the boundary zone of the infarction area was evaluated by TUNEL and Niss1 staining. The expressions of bFGF and GAP-43 and neurogenesis were evaluated by Western blotting and 5-bromodeoxyuridine (Brdu) fluorescence staining, respectively. After treatment with IA, the NSS of treatment groups were lower than that of the models (3 and 7 d). The number of TUNEL positive neurons decreased and Nissl positive neurons increased at the same time (3 d). The expressions of bFGF and GAP-43 increased significantly in the boundary zone of the infarction area when compared to model group. Moreover, IA markedly enhanced the neurogenesis in the brain after ischemia-reperfusion, which revealed an increase of Brdu/NeuN positive cells in the boundary zone of the infarction area. The possible mechanism of protecting neuronal injury of IA may be related to inhibition on neuronal apoptosis, upregulation of bFGF and GAP-43, and neurogenesis in boundary zone of infarction after cerebral ischemia-reperfusion.
Animals ; Apoptosis ; drug effects ; Brain Ischemia ; etiology ; Bromodeoxyuridine ; metabolism ; Fibroblast Growth Factor 2 ; metabolism ; GAP-43 Protein ; metabolism ; Infarction, Middle Cerebral Artery ; complications ; Male ; Neurogenesis ; drug effects ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Organic Chemicals ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; metabolism

OBJECTIVETo study the relationship between carotid atherosclerosis and cerebral infarction (CI).

METHODSBetween November 2008 and March 2009, 147 CI patients (CI group) and 48 patients with non-cerebrovascular diseases (control group) were enrolled from inpatients of Neurology Department of our hospital. The diagnostic criterion of thickened carotid intima was set as 1.0 mm

RESULTSIn the CI group, 36 (24.5%) patients had normal carotid intima, 22 (15.0%) had thickened carotid intima, and 89 (60.5%) had carotid plaque. In the control group, 22 (45.8%) patients had normal carotid intima, 4 (8.3%) had thickened carotid intima, and 22 (45.8%) had carotid plaque. The severity of carotid atherosclerosis in the CI group was higher than that in the control group (P = 0.022). There was significant difference in the constitution of carotid plaque between the two groups (P = 0.001); the CI group mainly had the soft plaque (55/89, 61.8%), whereas the control group mainly had the hard plaque (17/22, 77.3%). The first three common locations of carotid plaque in both groups were carotid bifurcation (CI group: 73.7%; control group: 64.1%), common carotid artery (CI group: 20.4%; control group: 25.6%), and internal carotid artery (CI group: 5.9%; control group: 10.3%). The location of carotid plaque between the two groups was not significantly different (P = 0.438). There was no difference in the carotid inner diameter or resistance index between the two groups (P > 0.05).

CONCLUSIONSCarotid atherosclerosis is to some extent able to reveal the atherosclerotic condition of cerebral arteries and act as an important predictor for the risk of CI. The color Doppler ultrasonography of carotid arteries can provide a convenient way for the prevention and treatment of CI.

Adult ; Aged ; Aged, 80 and over ; Carotid Arteries ; diagnostic imaging ; pathology ; Carotid Artery Diseases ; complications ; epidemiology ; pathology ; Cerebral Infarction ; epidemiology ; etiology ; pathology ; Female ; Humans ; Male ; Middle Aged ; Risk Factors ; Ultrasonography, Doppler, Color

The aim of the study is to investigate the effect of salvianolic acid B (SalB) on blood-brain barrier (BBB) in rats after cerebral ischemia-reperfusion, and to illustrate its possible mechanisms. Cerebral ischemia-reperfusion was induced by middle cerebral artery occlusion in rats. The break-down of BBB was indicated by extravasations of immunoglobulin (IgG) monitored with immunohistochemistry. The expression of MMP-9 and NOS2 in the brain was determined by immunohistochemistry, and the expression of p-p38 and p-ERK1/2 was detected by Western blotting. It was shown that on day 2 after ischemia-reperfusion the IgG accumulated around the vascular boundary zone, suggesting the break-down of BBB, and the expression of MMP-9 and NOS2 up-regulated at the same time. The result of Western blotting suggested that the expression of p-p38 and p-ERK1/2 increased. On day 7 after ischemia-reperfusion the. expression of MMP-9 and NOS2 was about the same level as day 2, the expression of p-p38 was higher than that on day 2 and the expression of p-ERK1/2 was slightly lower than that on day 2. SalB (1 and 10 mg x kg(-1)) significantly alleviated the extravasations of immunoglobulin induced by cerebral ischemia-reperfusion (P < 0.05). On day 2 and day 7 SalB attenuated the expression of MMP-9 and NOS2 (P < 0.05). SalB (10 mg x kg(-1)) reduced the expression of p-p38 and p-ERK1/2 apparently on day 2 and 7 after ischemia-reperfusion (P < 0.05). SalB (1 mg x kg(-1)) inhibited the expression of p-p38 on day 7 after ischemia-reperfusion (P < 0.05). The results indicate that SalB protects blood-brain barrier in rats after cerebral ischemia-reperfusion by inhibiting the MAPK pathway.
Animals ; Benzofurans ; isolation & purification ; pharmacology ; Blood-Brain Barrier ; drug effects ; metabolism ; Brain Ischemia ; etiology ; metabolism ; pathology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Infarction, Middle Cerebral Artery ; complications ; MAP Kinase Kinase Kinase 1 ; metabolism ; MAP Kinase Signaling System ; drug effects ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Phosphorylation ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology ; Salvia miltiorrhiza ; chemistry ; p38 Mitogen-Activated Protein Kinases ; metabolism

To detect the function and expression of ventricular M3 receptor (M3R) in cerebral-cardiac syndrome (CCS) model rats and to explore the relationship between the expression of M3R and the arrhythmia resulted from CCS, CCS model rats were induced by occluding right middle cerebral artery. ECG was monitored. Intracellular calcium ([Ca2+]i) changes after agitating M3R were recorded by laser scanning confocal microscope. Changes of M3R expression in the ventricular tissue were detected by Western blotting. QRS and QT intervals in CCS group were remarkably longer than that in sham group. According to the results of Western blotting, the level of M3R expression was remarkably lower in CCS group compared with that in the normal group. KCl induced [Ca2+]i increasing in CCS group could be depressed by choline and the effect of choline could be blocked by 4-DAMP. The lower expression of M3R in CCS group may be one of important reasons of arrhythmia resulted from CCS. M3R that depressed the [Ca2+]i increasing agitated by choline may become a new target to cure arrhythmia resulted from CCS.
Animals ; Arrhythmias, Cardiac ; etiology ; metabolism ; pathology ; physiopathology ; Calcium ; metabolism ; Choline ; pharmacology ; Electrocardiography ; Heart Ventricles ; metabolism ; Infarction, Middle Cerebral Artery ; complications ; Male ; Muscarinic Antagonists ; pharmacology ; Myocardium ; metabolism ; ultrastructure ; Myocytes, Cardiac ; metabolism ; Piperidines ; pharmacology ; Potassium Chloride ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Receptor, Muscarinic M3 ; antagonists & inhibitors ; metabolism

OBJECTIVETo investigate the activation of nuclear factor kappaB (NF-KB) and the influence of puerarin on it after cerebral ischemia-reperfusion in rats.

METHODCerebral ischemia-reperfusion injury was induced by 90 min of middle cerebral artery (MCA) occlusion and followed by 2, 6, 12, 24, 72 h reperfusion. Puerarin or saline was intra-peritoneally injected 1h before MCA occlusion and then the drugs were administered once every six hours. The infarct volume and brain edema were determined by TTC stain. Level of NF-kappaB P65 subunit was determined by immunohistochemistry and western blot.

RESULTImmunohistochemistry revealed the translocation of NF-kappaB. A time course of NF-kappaB induction in brain showed that NF-kappaB P65 subunit obviously increased at 6 h, peaked at 24 h and then decreased by 72 h post-reperfusion. Puerarin decreased the level of NF-kappaB at 24, 72 h after reperfusion. There was a decrease trend in brain infarct volume between puerarin and control.

CONCLUSIONNF-kappaB is translocated and its level is increased after ischemia-reperfusion. Puerarin may attenuate the ischemia-reperfusion injury through inhibition of NF-kappaB activation.

Animals ; Blotting, Western ; Brain ; drug effects ; metabolism ; pathology ; Immunohistochemistry ; Infarction, Middle Cerebral Artery ; complications ; Isoflavones ; isolation & purification ; pharmacology ; Male ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; physiopathology ; Transcription Factor RelA ; metabolism

OBJECTIVETo explore the effects of total saponins of Rubus parviflolius (TSRP) on brain edema and blood brain barrier in rats.

METHODThe model of local cerebral ischemia was established in rats by reversible inserting a nylon thread into the anterior cerebral artery through the internal carotid artery brain hydrated amount and content change of Evan' s blue (EB) in cortex subjected to 2h middle cererbral artery occlusion (MACO) followed by 6 h, 24 h, 48 h, 72 h reperfusion and effect of TSRP. penetrability of blood brain-barrier (BBB) the index includes brain hydrated amount and penetrability of blood brain-barrier BBB.

RESULTCom- pared with I/R group. Both brain hydrated amount and the EB content decreased significantly in TSRP groups on the 6 h, 24 h, 48 h, 72 h of reperfusion after 2 hour of cerebral ischemia induced by MACO model.

CONCLUSIONTSRP could decrease brain hydrated amount and markedly lower permeability of blood-brain barrier subjected to 2 h MACO followed by 24 h reperfusion, and this may be a mechanism of TSRP alleviating brain edema during I/R.

Animals ; Blood-Brain Barrier ; drug effects ; Brain Edema ; drug therapy ; etiology ; pathology ; Brain Ischemia ; complications ; Infarction, Middle Cerebral Artery ; complications ; Male ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; Rosaceae ; chemistry ; Saponins ; isolation & purification ; pharmacology

The aim of the present study was to assess the clinical and histopathological findings in a canine model of ischemic stroke. Cerebral ischemic stroke was induced by middle cerebral artery occlusion in four healthy beagle dogs using silicone plugs. They showed neurological signs of forebrain dysfunction such as reduced responsiveness, head turning, circling, postural reaction deficits, perceptual deficits, and hemianopsia. These signs gradually regressed within 4 weeks without therapy. On magnetic resonance imaging, T2 hyperintensity and T1 hypointensity were found in the cerebral cortex and basal ganglia. These lesions were well-defined and sharply demarcated from adjacent brain parenchyma with a homogenous appearance. No abnormalities of the cerebrospinal fluid were observed. At necropsy, atrophic and necrotic lesions were observed in the cerebral cortex. The cerebral cortex, basal ganglia, and thalamus were partially unstained with triphenyl-tetrazolium chloride. Histopathologically, typical features of infarction were identified in cortical and thalamic lesions. This study demonstrates that our canine model resembles the conditions of real stroke patients.
Animals ; Behavior, Animal/physiology ; Brain/metabolism/pathology ; Cerebral Infarction/*etiology/*pathology ; Cerebrospinal Fluid/chemistry/cytology ; Disease Models, Animal ; *Dogs ; Infarction, Middle Cerebral Artery/*complications/*pathology ; Magnetic Resonance Imaging ; Male