Permeability of the Blood-Brain Barrier within Rat Intrastriatal Transplants Assessed by Simultaneous Systemic Injection of Horseradish Peroxidase and Evans Blue Dye

Background and Purpose: Fear of symptomatic intracerebral hemorrhage (ICH) has been the primary reason for withholding tPA thrombolysis from acute ischemic stroke patients. Early blood brain barrier (BBB) damage is appreciated to be closely associated with post-thrombolysis ICH, while it remains a technical challenge for rapid assessment of BBB damage before tPA administration. Our recent data showed that cerebral ischemia induced rapid degradation of tight junction protein occludin in ischemic cerebromicrovessels. This study further investigates whether the cleaved occludin is released into the blood stream and how blood occludin levels correlate to the extent of ischemic BBB damage. Methods: Male Sprague Dawley rats were subjected to 1.5, 3, 4.5, 12 and 24 hours of middle cerebral artery occlusion (MCAO), followed by 5-min reperfusion. Blood samples were taken before and after MCAO. Blood occludin was assessed by ELISA. BBB permeability was measured by Evans blue dye leakage. Occludin cleavage was identified on immunoblots. Results: MCAO induced Evans blue dye leakage and blood occludin increase in a duration-dependent manner. Blood occludin increase concurrently occurred with the loss of occludin from ischemic cerebral microvessels. Western blot analysis identified two cleaved occludin fragments (31- and 55- kDa) in the blood. Lastly, blood occludin levels remained significantly higher than its basal level within the first 24 hours after MCAO onset. Conclusions: Our results indicate that blood occludin levels correlate well with the extents of BBB damage and thus may serve as a potential biomarker for evaluating the risk of hemorrhagic transformation before tPA administration.

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The influence of nimodipine on cerebral blood flow autoregulation and blood-brain barrier

Journal of Neurosurgery ◽

10.3171/jns.1988.69.6.0919 ◽

1988 ◽

Vol 69 (6) ◽

pp. 919-922 ◽

Cited By ~ 34

Author(s):

Hans-Georg Höllerhage ◽

Michael R. Gaab ◽

Matthias Zumkeller ◽

Gerhard F. Walter

Keyword(s):

Blood Flow ◽

Cerebral Blood Flow ◽

Blood Brain Barrier ◽

Evans Blue ◽

Brain Barrier ◽

Control Group ◽

Blue Dye ◽

Evans Blue Dye ◽

Blood Brain ◽

Anesthetized Rats

✓ Twenty anesthetized rats were randomly assigned to a nimodipine-treated group or a control group of 10 rats each. Local cerebral blood flow (lCBF) was measured by means of a surface electrode using the hydrogen clearance technique. Systemic arterial pressure (SAP) was varied with administration of norfenefrine or by hemorrhage in order to obtain SAP/cerebral blood flow (CBF) curves under different conditions. In the control group, a typical autoregulation curve was obtained with an lCBF plateau between 70 and 120 mm Hg SAP. The nimodipine-treated animals, however, showed only a slight diminution in the slope of the curve but no real plateau, indicating impairment of CBF autoregulation. In another series, 20 anesthetized rats were randomly assigned to a treatment group or a control group of 10 animals each. Intravenous Evans blue dye was used as a tracer for blood-brain barrier (BBB) function. In both groups, SAP was raised to a level of 180 mm Hg with administration of norfenefrine for 6 minutes. Extravasation of significantly more Evans blue dye was observed in the nimodipine group than in the control group, indicating impairment of the BBB. It is concluded that nimodipine may impair CBF autoregulation, allowing damage to the BBB under hypertensive conditions.

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Evaluation of Blood-Brain Barrier Integrity Using Vascular Permeability Markers: Evans Blue, Sodium Fluorescein, Albumin-Alexa Fluor Conjugates, and Horseradish Peroxidase

10.1007/7651_2020_316 ◽

2020 ◽

Author(s):

Bulent Ahishali ◽

Mehmet Kaya

Keyword(s):

Horseradish Peroxidase ◽

Vascular Permeability ◽

Blood Brain Barrier ◽

Evans Blue ◽

Brain Barrier ◽

Sodium Fluorescein ◽

Barrier Integrity ◽

Alexa Fluor ◽

Blood Brain ◽

Blood Brain Barrier Integrity

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A Study of the Pathogenesis and Prevention of Central Pontine Myelinolysis in a Rat Model

Journal of International Medical Research ◽

10.1177/147323000603400305 ◽

2006 ◽

Vol 34 (3) ◽

pp. 264-271 ◽

Cited By ~ 6

Author(s):

Q-H Ke ◽

T-B Liang ◽

J Yu ◽

S-S Zheng

Keyword(s):

Hypertonic Saline ◽

Blood Brain Barrier ◽

Evans Blue ◽

Central Pontine Myelinolysis ◽

Brain Barrier ◽

Blue Dye ◽

Evans Blue Dye ◽

Pontine Myelinolysis ◽

Demyelinating Lesions ◽

Central Pontine

The development of central pontine myelinolysis was studied in rats. Severe hyponatraemia was induced using vasopressin tannate and 2.5% dextrose in water and then rapidly corrected with hypertonic saline alone, hypertonic saline and dexamethasone simultaneously, or hypertonic saline plus dexamethasone 24 h later. The permeability of the blood-brain barrier was evaluated using the extravasation of Evans blue dye and the expression of inducible nitric oxide synthase (iNOS) in the brain was examined using Western blot analysis. Histological sections were examined for demyelinating lesions. In rats receiving hypertonic saline alone, Evans blue dye content and expression of iNOS began to increase 6 and 3 h, respectively, after rapid correction of hyponatraemia and demyelinating lesions were seen. When dexamethasone was given simultaneously with hypertonic saline, these increases were inhibited and demyelinating lesions were absent. These effects were lost if dexamethasone injection was delayed. Disruption of the blood-brain barrier and increased iNOS expression may be involved in the pathogenesis of central pontine myelinolysis, and early treatment with dexamethasone may help prevent the development of central pontine myelinolysis.

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The development of the blood-brain barrier in the chick. Studies with Evans blue and horseradish peroxidase

Annals of Anatomy - Anatomischer Anzeiger ◽

10.1016/s0940-9602(11)80247-5 ◽

1993 ◽

Vol 175 (1) ◽

pp. 85-88 ◽

Cited By ~ 10

Author(s):

D. Ribatti ◽

B. Nico ◽

M. Bertossi

Keyword(s):

Horseradish Peroxidase ◽

Blood Brain Barrier ◽

Evans Blue ◽

Brain Barrier ◽

Blood Brain

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The influence of systemic arterial pressure and intracranial pressure on the development of cerebral vasogenic edema

Journal of Neurosurgery ◽

10.3171/jns.1983.59.5.0803 ◽

1983 ◽

Vol 59 (5) ◽

pp. 803-809 ◽

Cited By ~ 82

Author(s):

Quentin J. Durward ◽

Rolando F. Del Maestro ◽

A. Loren Amacher ◽

J. Keith Farrar

Keyword(s):

Blood Pressure ◽

Arterial Pressure ◽

Blood Brain Barrier ◽

Evans Blue ◽

Vasogenic Edema ◽

Brain Barrier ◽

Blue Dye ◽

Content Type ◽

Evans Blue Dye ◽

Fine Print

✓ The influence of intracranial pressure (ICP), systemic arterial pressure (SAP), and cerebral perfusion pressure (CPP) upon the development of vasogenic cerebral edema is largely unknown. To study their relationship, the authors have produced an osmotic disruption of the blood-brain barrier unilaterally in rabbits by injecting 1 cc/kg of 2M NaCl into the left internal carotid artery. The amount of vasogenic edema produced was assessed by quantitation of the extravasation of Evans blue dye into the area of maximum blood-brain barrier breakdown by means of optical densitometry following formamide extraction. The ICP was measured using a cisterna magna catheter into which mock cerebrospinal fluid could be infused at a predetermined pressure. The SAP was controlled by exsanguination from a femoral artery catheter. In 18 animals in which blood pressure was not controlled, no significant relationship between the ICP and the degree of Evans blue dye extravasation was noted. In these animals, however, a direct relationship between CPP (defined as mean arterial pressure minus mean ICP) and extravasation of Evans blue dye was found (correlation coefficient 0.630; p < 0.001). When ICP was held constant at 0 to 5 mm Hg in another group of 16 animals and different levels of blood pressure were produced by exsanguination, a significant direct relationship between extravasation of Evans blue dye and the SAP was found (correlation coefficient 0.786; p < 0.001). In a third group of 20 animals, the blood pressure was held constant at 90 to 100 mm Hg and the ICP was varied between 0 and 75 mm Hg. There was a highly significant result indicating increasing Evans blue dye extravasation with lower levels of ICP (p < 0.001). Cerebral blood flow determinations by the hydrogen clearance method indicated loss of autoregulation in all animals in the areas of brain injured by intracarotid hypertonic saline. These results indicate that high SAP and low ICP (that is, a large CPP) promote Evans blue dye extravasation in this model of blood-brain barrier disruption. This finding has implications for the management of patients with vasogenic edema.

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