Cellular and secretory mechanisms related to delayed radiation-induced microvessel dysfunction in the spinal cord of rats.
OBJECTIVE
This study aimed to investigate long-term, radiation-induced changes in microvessel permeability, the profile of the vasoactive mediators endothelin and nitric oxide, and the response of specific cell systems in the irradiated spinal cord of rats.


METHODS
The thoracolumbar spinal cords of Fischer rats were irradiated to a dose of 15 Gy, and the rats were sacrificed at various times afterward. Endothelin levels and nitric oxide-synthase (NOS) activity were assayed in extracts of spinal cords. Microvascular permeability and the effect of treatment with recombinant human manganese superoxide dismutase (r-hMnSOD) were assessed quantitatively. Immunohistochemistry evaluated astrocytes, microglia, vascular basal membrane, and neurofilaments.


RESULTS
None of the rats developed neurologic dysfunction. Endothelin levels were significantly reduced at 18 h after irradiation and markedly attenuated after 10 days (p < 0.007). Thereafter, endothelin levels returned to normal values at 56 days after radiation and escalated to markedly high levels after 120 and 180 days (p < 0.002). NOS activity remained very low throughout the period of follow-up and failed to counterbalance the shifts in endothelin levels. Treatment with r-hMnSOD had no effect on normal vascular permeability but it abolished the abnormally increased permeability measured at 18 h after radiation and again after 120 and 180 days. Standard microscopic evaluation failed to reveal abnormalities in the irradiated spinal cord, but immunohistochemical staining showed a progressive increase in the number of microglial cells per field after 120 and 180 days (p < 0003). A similar increase in the number of astrocytic cells per field was noted after more than 180 days, but an earlier short lasting peak was also noted at 14 days after radiation. No abnormalities were found in blood vessel configuration, density, diameter, and basal membrane staining, or in the neurofilaments.


CONCLUSIONS
Marked imbalance in the regulatory function of endothelium-derived mediators of the vascular tone is present after radiation therapy probably inducing chronic vasoconstriction. This imbalance favors localized procoagulation that may enhance the consequent loss of function measured as increased permeability. Microglial proliferation may account for continuous release of superoxide that may enhance disruption of normal permeability. The latter is corrected by SOD treatment. Astrocytic proliferation may present a response to the mitogenic effect of endothelin and to microglial-derived paracrine effect of cytokines.
Siegal
T
T
Department of Neurology, Hadassah Hebrew University Hospital, Jerusalem, Israel. Siegal@hadassah.org.il
Pfeffer
M R
MR
Meltzer
A
A
Shezen
E
E
Nimrod
A
A
Ezov
N
N
Ovadia
H
H
D000818:Animals; D002199:Capillary Permeability / Q000187:drug effects / Q000528:radiation effects*; D016232:Endothelins / Q000378:metabolism*; D005260:Female; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D007150:Immunohistochemistry; D009474:Neurons / Q000737:chemistry; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D051381:Rats; D011916:Rats, Inbred F344; D013116:Spinal Cord / Q000098:blood supply / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects*; D013482:Superoxide Dismutase / Q000494:pharmacology; D013997:Time Factors
D016232:Endothelins; D005904:Glial Fibrillary Acidic Protein; D019001:Nitric Oxide Synthase; D013482:Superoxide Dismutase