Glutathione supplementation to University of Wisconsin solution causes endothelial dysfunction.
BACKGROUND
Glutathione (GSH) is added to University of Wisconsin (UW) organ preservation solution to protect against oxidative stress. This study assesses the effect of GSH-supplementation on endothelial function in tissues subjected to cold ischaemia and compares its effects to a mono-ethyl ester equivalent (GSH-MEE) and S-nitrosated GSH (GSNO).


METHODS
Rat aortic rings were stored for 1 h or 48 h in cold, hypoxic UW solution with or without GSH (3 mM), GSH-MEE (3 mM) or GSNO (100 mciroM) supplementation. Aortic rings were reoxygenated in warm Krebs solution; smooth muscle function was assessed by responses to phenylephrine (PE), and endothelial function by vasodilatation to the endothelium-dependent dilator, acetylcholine (ACh). The protective effects against oxidant-induced endothelial cell death were assessed in cultured human umbilical vein endothelial cells (HUVEC).


RESULTS
Supplementation of UW with either GSH or GSH-MEE had no effect on vascular responses to PE, but smooth muscle contraction was significantly attenuated in rings incubated for 48 h with GSNO. Endothelium-dependent relaxation was significantly impaired in tissues stored under hypoxic conditions in GSH, GSH-MEE and GSNO supplemented UW solution for 1 h. However, impairment at 48 h was significantly more pronounced in GSH-treated vessels. Cultured HUVEC death was exacerbated by GSH and GSH-MEE in unstressed cells and in those stressed with a superoxide anion generator.


CONCLUSIONS
GSH supplementation of UW solution exacerbates cold-ischaemia induced endothelial dysfunction. GSNO did not share the detrimental effects of GSH and promoted NO-mediated vasodilatation.
Tambyraja
A L
AL
Clinical & Surgical Sciences (Surgery), Royal Infirmary of Edinburgh, 51 Little France Crescent, University of Edinburgh, Edinburgh, EH16 4SA, UK. andrew.tambyraja@ed.ac.uk
Mitchell
R
R
Driscoll
P J
PJ
Deans
C
C
Parks
R W
RW
Rahman
I
I
Megson
I L
IL
D000241:Adenosine / Q000494:pharmacology; D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D050377:Cold Ischemia / Q000379:methods*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D005978:Glutathione / Q000494:pharmacology*; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D008297:Male; D019188:Organ Preservation Solutions / Q000494:pharmacology*; D018384:Oxidative Stress; D011887:Raffinose / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D026422:S-Nitrosoglutathione / Q000097:blood / Q000494:pharmacology
D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D026422:S-Nitrosoglutathione; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose