Neuroprotective effects of recombinant thrombomodulin in controlled contusion spinal cord injury implicates thrombin signaling.
Although the central nervous system (CNS) of mammals has had poor prospects for regeneration, recent studies suggest this might improve from blocking "secondary cell loss" or apoptosis. In this regard, intravenous activated protein C (aPC) improved neurologic outcomes in a rat compression spinal cord injury (SCI) model. Protein C activation occurs when the serine protease thrombin binds to the cell surface proteoglycan thrombomodulin (TM) forming a complex that halts coagulation. In culture, rTM blocks thrombin's activation of protease-activated receptors (PARs), that mediate thrombin killing of neurons and glial reactivity. Both PAR1 and prothrombin are rapidly upregulated after contusion SCI in rats, prior to peak apoptosis. We now report neuroprotective effects of intraperitoneal soluble recombinant human rTM on open-field locomotor rating scale (BBB) and spinal cord lesion volume when given 1 h after SCI. BBB scores from four separate experiments showed a 7.6 +/- 1.4 absolute score increase (p < 0.05) at 3 days, that lasted throughout the time course. Histological sections at 14 days were even more dramatic where a twofold reduction in lesion volume was quantified in rTM-treated rats. Thionin staining revealed significant preservation of motor neuronal profiles both at, and two segments below, the lesion epicenter. Activated caspase-3 immunocytochemistry indicated apoptosis was quite prominent in motor neurons in vehicle (saline) controls, but was dramatically reduced by rTM. Microglia, increased and activated after injury, were reduced with rTM treatment. Taken together, these and previous results support a prominent role for coagulation-inflammation signaling cascades in the subacute changes following SCI. They identify a neuroprotective role for rTM by its inhibition of thrombin generation and blockade of PAR activation.
Neurobiology Research Laboratory, Heartland Network, Department of Veterans Affairs Medical Center, Kansas City, Missouri 64128, USA. bfestoff@kumc.edu
Barry W
BW
Festoff
Syed
S
Ameenuddin
Karen
K
Santacruz
John
J
Morser
Zhiming
Z
Suo
Paul M
PM
Arnold
Kristie E
KE
Stricker
Bruce A
BA
Citron
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D005260:Female; D007150:Immunohistochemistry; D009043:Motor Activity / Q000187:drug effects; D018696:Neuroprotective Agents / Q000627:therapeutic use*; D010314:Partial Thromboplastin Time; D051381:Rats; D017207:Rats, Sprague-Dawley; D044463:Receptor, PAR-1 / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins / Q000627:therapeutic use; D020127:Recovery of Function; D015398:Signal Transduction / Q000502:physiology*; D013119:Spinal Cord Injuries / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D013917:Thrombin / Q000187:drug effects / Q000378:metabolism*; D018180:Thrombomodulin / Q000627:therapeutic use*
D018696:Neuroprotective Agents; D044463:Receptor, PAR-1; D011994:Recombinant Proteins; D018180:Thrombomodulin; D013917:Thrombin