Intra-ductal glutamine administration reduces oxidative injury during human pancreatic islet isolation.
Oxidative stress during islet isolation induces a cascade of events injuring islets and hampering islet engraftment. This study evaluated islet isolation and transplantation outcomes after intra-ductal glutamine administration. Human pancreata deemed unsuitable for pancreas or islet transplantation were treated with either a 5 mM solution of l-glutamine (n = 6) or collagenase enzyme alone (n = 6) through the main pancreatic duct. Islet yield, viability, in vitro function; markers of oxidative stress [malondialdehyde (MDA) and Glutathione (GSH)] and apoptosis were assessed. Islet yields were significantly increased in the glutamine group compared to controls (318, 559 +/- 25, 800 vs. 165, 582 +/- 39, 944 mean +/- SEM, p < 0.01). The amount of apoptotic cells per islet was smaller in the glutamine group than the control. The percentage of nude mice rendered normoglycemic with glutamine-treated islets was higher than the controls (83% n = 10/12 vs. 26% n = 6/23; p < 0.01), and the time to reach normoglycemia was decreased in the glutamine group (1.83 +/- 0.4 vs. 7.3 +/- 3 days; p < 0.01). Glutamine administration increased GSH levels (7.6 +/- 1.7 nmol/mg protein vs. 4.03 +/- 0.5 in control, p < 0.05) and reduced lipid-peroxidation (MDA 2.45 +/- 0.7 nmol/mg of protein vs. 6.54 +/- 1.7 in control; p < 0.05). We conclude that intra-ductal administration of glutamine reduces oxidative injury and apoptosis and improves islet yield and islet graft function after transplantation.
Avila
J
J
University of Illinois at Chicago, Division of Transplantation, Chicago, USA.
Barbaro
B
B
Gangemi
A
A
Romagnoli
T
T
Kuechle
J
J
Hansen
M
M
Shapiro
J
J
Testa
G
G
Sankary
H
H
Benedetti
E
E
Lakey
J
J
Oberholzer
J
J
D000328:Adult; D000368:Aged; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism / Q000601:surgery*; D005260:Female; D005973:Glutamine / Q000494:pharmacology*; D005978:Glutathione / Q000378:metabolism; D006085:Graft Survival; D006801:Humans; D020287:In Situ Nick-End Labeling; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D016381:Islets of Langerhans Transplantation; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D018384:Oxidative Stress / Q000187:drug effects*; D010183:Pancreatic Ducts; D010477:Perfusion; D014183:Transplantation, Heterologous
D005973:Glutamine; D008315:Malondialdehyde; D005978:Glutathione