A new model of cerebral microthrombosis in rats and the neuroprotective effect of a Rho-kinase inhibitor.
OBJECTIVE
The aim of this study was to develop a new model of stroke based on endothelial damage and thrombotic occlusion in a perforating artery, leading to small cerebral infarcts and neurological deficits in rats. Moreover, the neuroprotective efficacy of fasudil, a rho-kinase inhibitor, was investigated in this model.


METHODS
Fifty-six male Sprague-Dawley rats were used in the present study. Rats were anesthetized with sodium pentobarbital, and 100 microg of sodium laurate was injected into the left internal carotid artery on days 1 and 3. The thrombus induction and consequent of ischemic brain damage were examined by histopathological analyses and neurological deficit scoring in a posture reflex test. To investigate the neuroprotective effects of fasudil, 1 or 10 mg/kg was administered intraperitoneally 5 minutes after the first injection of sodium laurate and once daily thereafter on the following 2 days.


RESULTS
One hour after the injection of sodium laurate, microscopic examination of phosphotungstic acid hematoxylin-stained sections (n=5) revealed that microthrombi containing fibrin strands obstructed the perforating arteries in the ipsilateral hemisphere. Under a transmission electron microscope (n=6), endothelial cells appeared exfoliated and the vascular lumen was obstructed by a thrombus composed of degranulated platelets, fibrin, leukocytes, and erythrocytes. No evidence of endothelial cell damage or thrombus could be found in the ipsilateral side of the pial artery (middle cerebral artery). Twenty-four hours after the second injection of sodium laurate (day 4), 13 of 15 rats (86.6%) showed mild to severe neurological deficits. Multiple small cerebral infarcts were observed in the hippocampus, cortex, and thalamus. Treatment with fasudil (1 and 10 mg/kg, n=15 each) resulted in a significant improvement in neurological deficits. Fasudil also significantly reduced the area of cerebral infarction.


CONCLUSIONS
We present a new model of stroke in rats, in which the perforating arteries are selectively occluded by microthrombi. This model is useful to investigate the pathophysiology and treatment of small cerebral infarction, which is caused by perforating arterial occlusive diseases such as lacunar infarcts. Fasudil may be beneficial in the treatment of acute ischemic stroke.
Toshima
Y
Y
Laboratory for Pharmacology, Asahi Chemical Industry, Shizuoka, Japan. a9187022@ut.asahi-kasei.co.jp
Satoh
S
S
Ikegaki
I
I
Asano
T
T
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives / Q000627:therapeutic use*; D000818:Animals; D020520:Brain Infarction / Q000188:drug therapy / Q000503:physiopathology; D002343:Carotid Artery, Internal / Q000473:pathology; D002540:Cerebral Cortex / Q000473:pathology; D004195:Disease Models, Animal; D004730:Endothelium, Vascular / Q000473:pathology / Q000648:ultrastructure; D004791:Enzyme Inhibitors / Q000627:therapeutic use; D006624:Hippocampus / Q000473:pathology; D047908:Intracellular Signaling Peptides and Proteins; D007850:Lauric Acids; D008297:Male; D008854:Microscopy, Electron; D009460:Neurologic Examination; D018696:Neuroprotective Agents / Q000627:therapeutic use*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012018:Reflex / Q000187:drug effects; D013194:Staining and Labeling; D020521:Stroke / Q000139:chemically induced / Q000188:drug therapy* / Q000503:physiopathology; D013788:Thalamus / Q000473:pathology; D054460:rho-Associated Kinases
D004791:Enzyme Inhibitors; D047908:Intracellular Signaling Peptides and Proteins; D007850:Lauric Acids; D018696:Neuroprotective Agents; C030358:lauric acid; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases; C049347:fasudil