Brain phospholipase A2 is activated after experimental closed head injury in the rat.
Head injury was induced in rats by a weight drop device, falling over the left hemisphere. The rats were killed at 15 min, 4 h, and 24 h after injury. Cortical slices were taken from the injured zone, from the corresponding region of the contralateral hemisphere, and from the frontal lobe of both hemispheres. These cortical slices were incubated in the presence of a fluorescent phospholipid analogue, 1-acyl-2-(N-4-nitrobenzo-2-oxa-1,3-diazole)aminocaproylphosphatidylch oli ne (C6-NBD-PC) which is a substrate for phospholipase A2 (PLA2) in intact cells. The interaction of this substrate with cells produces only one fluorescent product, the fatty acid C6-NBD-FA, released from the 2-position of C6-NBD-PC. Thus, the level of C6-NBD-FA produced is a direct measure of PLA2 activity. Fifteen minutes after trauma, a 75% increase of PLA2 activity was found in the injured zone. At 4 h, the frontal lobe of the contused, left hemisphere had elevated PLA2 activity, as well as the injured zone (92 and 81%, respectively). At 24 h, PLA2 activity at the site of injury was 245% of sham. In the right, noninjured zone, no significant changes in PLA2 activity were noticed during the entire time course of the experiment. Prostaglandin E2 (PGE2) was extracted from the same cortical slices as those used for PLA2 activity measurement.(ABSTRACT TRUNCATED AT 250 WORDS)
Shohami
E
E
Department of Pharmacology, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Shapira
Y
Y
Yadid
G
G
Reisfeld
N
N
Yedgar
S
S
D000818:Animals; D001809:Blood Viscosity / Q000187:drug effects; D001921:Brain / Q000201:enzymology*; D006259:Craniocerebral Trauma / Q000201:enzymology*; D003911:Dextrans / Q000494:pharmacology; D015232:Dinoprostone / Q000378:metabolism; D004789:Enzyme Activation; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D008297:Male; D010713:Phosphatidylcholines; D010740:Phospholipases / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D051381:Rats; D011919:Rats, Inbred Strains; D013929:Thromboxane B2 / Q000378:metabolism; D013997:Time Factors
D003911:Dextrans; D005456:Fluorescent Dyes; D010713:Phosphatidylcholines; D013929:Thromboxane B2; C044745:1-palmitoyl-2-(6-(N-(7-nitrobenz)-2-oxa-1,3-diazol-4-yl)aminocaproyl)phosphatidylcholine; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D015232:Dinoprostone