The metabotropic glutamate receptor system protects against ischemic free radical programmed cell death in rat brain endothelial cells.
As one of the key determinants of ischemic injury, cerebrovascular endothelial cell (EC) degeneration may be dependent upon the generation of the free radical nitric oxide (NO) and the subsequent induction of programmed cell death (PCD). Although the mechanisms that can prevent EC injury are most likely multifactorial in origin, the metabotropic glutamate receptor (mGluR) system may represent a novel therapeutic approach for ECs given the ability of the mGluR system to reverse neuronal cell injury. This study examined the modulation of individual subtypes of mGluRs during anoxia and NO toxicity in primary rat cerebrovascular ECs. Cell injury was determined through trypan blue dye exclusion, intracellular lactate dehydrogenase release, DNA fragmentation, membrane phosphatidylserine (PS) exposure, and cysteine protease activity. Anoxia, through the generation of NO, and exposure to exogenous NO were directly toxic to ECs. Exposure to NO rapidly decreased EC viability from 98% +/- 2% to 40% +/- 9%, increased DNA fragmentation from 2% +/- 2% to 61% +/- 9%, and increased membrane PS exposure from 3% +/- 3% to 66% +/- 6% over a 24-hour period. Activation of the mGluR system significantly increased EC survival through the prevention of NO-induced DNA fragmentation and cellular membrane PS residue exposure. In contrast, antagonism of the mGluR system failed to prevent PCD. Cytoprotection by the mGluR system was dependent, at least in part, upon the direct inhibition of NO-generated caspase 1- and caspase 3-like activities. Further investigation into the ability of the mGluR system to prevent PCD in ECs may open new therapeutic avenues for the treatment of cerebrovascular injury.
Lin
S H
SH
Laboratory of Cellular and Molecular Cerebral Ischemia, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
Maiese
K
K
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001921:Brain / Q000098:blood supply / Q000378:metabolism / Q000473:pathology; D020170:Caspase 1 / Q000378:metabolism; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000378:metabolism; D053938:DNA Fragmentation; D004730:Endothelium, Vascular / Q000201:enzymology / Q000473:pathology; D004789:Enzyme Activation / Q000502:physiology; D005609:Free Radicals / Q000378:metabolism; D020925:Hypoxia-Ischemia, Brain / Q000378:metabolism* / Q000473:pathology*; D020287:In Situ Nick-End Labeling; D009569:Nitric Oxide / Q000096:biosynthesis; D010718:Phosphatidylserines / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D018094:Receptors, Metabotropic Glutamate / Q000378:metabolism*
D005609:Free Radicals; D010718:Phosphatidylserines; D018094:Receptors, Metabotropic Glutamate; D009569:Nitric Oxide; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases; D020170:Caspase 1