title: Neuroprotection from secondary injury by polyethylene glycol requires its internalization.
abstract: Polyethylene glycol (PEG) is well known to both fuse and repair cell membranes. This capability has been exploited for such diverse usages as the construction of hybridomas and as a reparative agent following neurotrauma. The latter development has proceeded through preclinical testing in cases of naturally induced paraplegia in dogs. The mechanisms of action of polymer-mediated neurorepair/neuroprotection are still under investigation. It is likely that the unique interaction of hydrophilic polymers with the mechanical properties of cell membranes in concert with an ability to interfere with mechanisms of secondary injury such as the production of highly reactive oxygen species (ROS or ;free radicals') is the basis for neuroprotection by polymers. Here we provide further evidence that the ability of PEG to reduce or limit secondary injury and/or lipid peroxidation (LPO) of membranes requires entry of PEG into the cytosol, further suggesting a physical interaction with the membranes of organelles such as mitochondria as the initial event leading to neurorepair/neuroprotection. We have evaluated this relationship in vitro using acrolein, a potent endogenous toxin that is a product of LPO. Acrolein can pass through cell membranes with ease, inducing progressive LPO in ;bystander' cells, and the production of even more acrolein by inducing its own production. Immediate application of PEG (10 mmol l(-1), 2000 Da) to poisoned neurons in vitro was unable to rescue them from necrosis and death. Furthermore, three-dimensional confocal microscopy of fluorescently decorated PEG shows that it does not enter these cells for up to 2 h after application. By this time the mechanisms of necrosis are likely irreversible. Additionally, severe oxygen and or glucose deprivation of spinal cord white matter in vitro also initiates LPO. Addition of potent free radical scavengers such as ascorbic acid or superoxide dismutase (SOD) is able to interfere with this process, but PEG is not. Taken together, these data are consistent with the hypothesis that PEG is able to rescue mechanically damaged cells, based on a restructuring of the damaged plasmalemma. Furthermore, in compromised cells with an intact cell membrane, PEG must first gain access to the cytosol where this same capability may be useful in restoring the integrity of cellular organelles such as mitochondria, though the intracellular concentration of the polymer must be significant relative to the concentration of toxins produced by LPO in order to rescue the cell.
authors.affiliation: Center for Paralysis Research, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: Peishan
authors.initials: P
authors.lastname: Liu-Snyder
authors.forename: Melissa Peasley
authors.initials: MP
authors.lastname: Logan
authors.forename: Riyi
authors.initials: R
authors.lastname: Shi
authors.forename: Daniel T
authors.initials: DT
authors.lastname: Smith
authors.forename: Richard Ben
authors.initials: RB
authors.lastname: Borgens
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000166:cytology*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D016716:PC12 Cells; D011092:Polyethylene Glycols / Q000494:pharmacology*; D051381:Rats; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D018696:Neuroprotective Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; D011092:Polyethylene Glycols; C022616:thiazolyl blue