Effect of thrombin on blood brain barrier permeability and its mechanism.
BACKGROUND
Previous studies have indicated that thrombin (TM) may play a major role in brain edema after intracerebral hemorrhages (ICHs). However, the mechanism of TM-induced brain edema is poorly understood. In this study, we explored the effect of TM on the permeability of the blood brain barrier (BBB) and investigated its possible mechanism, aiming at providing a potential target for brain edema therapy after ICHs.


METHODS
TM or TM + cathepsin G (CATG) was stereotaxically injected into the right caudate nucleus of Sprague-Dawley rats in vivo. BBB permeability was measured by Evans-Blue extravasation. Brain water content was determined by the dry-wet weight method. Brain microvascular endothelial cells were then cultured in vitro. After TM or TM + CATG was added to the endothelial cell medium, changes in the morphology of cells were dynamically observed by phase-contrast light microscopy, and the expression of matrix metalloproteinase-2 (MMP-2) protein was measured by immunohistochemical method.


RESULTS
BBB permeability increased at 6 hours after a TM injection into the ipsilateral caudate nucleus (P < 0.05), peaked between 24 hours (P < 0.01) and 48 hours (P < 0.05) after the injection, and then declined. Brain water content changed in parallel with the changes in BBB permeability. However, at all time points, BBB permeability and brain water content after a TM + CATG injection were not significantly different from the respective parameters in the control group (P > 0.05). TM induced endothelial cell contraction in vitro in a time-dependent manner and enhanced the expression of MMP-2 protein. After incubation with TM + CATG, cell morphology and MMP-2 expression did not change significantly as compared to the control group (P > 0.05).


CONCLUSIONS
Increased BBB permeability may be one of the mechanisms behind TM-induced cerebral edema. TM induces endothelial cell contraction and promotes MMP-2 expression by activating protease activated receptor-1 (PAR-1), possibly leading to the opening of the BBB.
Guan
Jing-Xia
JX
Department of Neurology, Union Medical College Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China. tsinhust@sohu.com
Sun
Sheng-Gang
SG
Cao
Xue-Bing
XB
Chen
Zhi-Bin
ZB
Tong
E-Tang
ET
D000818:Animals; D001812:Blood-Brain Barrier / Q000187:drug effects*; D001834:Body Water / Q000378:metabolism; D001929:Brain Edema / Q000209:etiology; D056649:Cathepsin G; D002403:Cathepsins / Q000494:pharmacology; D002543:Cerebral Hemorrhage / Q000150:complications*; D020778:Matrix Metalloproteinase 2 / Q000032:analysis; D010539:Permeability; D051381:Rats; D017207:Rats, Sprague-Dawley; D044463:Receptor, PAR-1 / Q000502:physiology; D012697:Serine Endopeptidases; D013917:Thrombin / Q000633:toxicity*
D044463:Receptor, PAR-1; D002403:Cathepsins; D012697:Serine Endopeptidases; D056649:Cathepsin G; C534097:Ctsg protein, rat; D013917:Thrombin; D020778:Matrix Metalloproteinase 2