Glutamine infusion during ischemia is detrimental in a murine gut ischemia/reperfusion model.
BACKGROUND
Gut ischemia/reperfusion (I/R) frequently occurs in clinical settings as a result of disproportionate splanchnic hypoperfusion during shock. Glutamine (GLN) supplementation of total parenteral nutrition (TPN) before gut I/R improves survival after gut I/R compared with standard TPN. However, it is unknown whether GLN treatment after the occurrence of the insult is beneficial or not. The aims of this study were to examine effects of GLN infusion during gut ischemia on survival, myeloid cell (neutrophils + monocytes) activation, and vascular permeability in organs.


METHODS
Male Institute of Cancer Research (ICR) mice were randomized to control and GLN groups. After IV cannulation, mice underwent 90 (experiments 1 and 2) or 60 (experiment 3) minutes of gut I/R. Control mice received normal saline infusion at 1 mL/h for 60 minutes during ischemia, whereas the GLN group was given 3% GLN solution. In experiment 1, survival rates were monitored for 72 hours (n = 25). In experiment 2, peripheral blood was obtained at 2 or 4 hours after reperfusion (n = 17). Reactive oxygen intermediate (ROI) production by myeloid cells was determined by flow cytometry using dihydrorhodamine 123 with or without phorbol myristate acetate stimulation. Expression of CD11a and CD11b on myeloid cells was also measured. Myeloperoxidase (MPO) activity in the lung was evaluated. In experiment 3, vascular permeability in organs was measured using Evans blue at 2 or 4 hours.


RESULTS
In experiment 1, survival time in the GLN group was significantly reduced compared with the control group (p = .02, log-rank test). The survival rates were 92% (12/13) and 42% (5/12) for the control and GLN groups at 12 hours (p = .01) and 38% (5/13) and 0% (0/12) at 48 hours (p = .02), respectively. In experiment 2, ROI production was significantly higher in the GLN group than in the control group after PMA stimulation both at 2 and 4 hours. CD11b expression was significantly higher in the GLN group than in the control group at 4 hours. There was no difference in pulmonary MPO activity at either time point. In experiment 3, GLN infusion significantly increased hepatic vascular permeability compared with saline infusion at 4 hours.


CONCLUSIONS
GLN infusion during ischemia is detrimental for survival after gut I/R. A possible mechanism is excessive priming of myeloid cells caused by GLN infusion. Timing of GLN administration is critical for outcome after gut ischemic insult.
Fukatsu
Kazuhiko
K
Division of Basic Traumatology, National Defense Medical College Research Institute, Tokorozawa-city, Saitama, Japan. kazfukatsu@yahoo.co.jp
Ueno
Chikara
C
Hashiguchi
Yojiro
Y
Hara
Etsuko
E
Kinoshita
Manabu
M
Mochizuki
Hidetaka
H
Hiraide
Hoshio
H
D000818:Animals; D002199:Capillary Permeability / Q000187:drug effects; D005973:Glutamine / Q000008:administration & dosage / Q000633:toxicity*; D007262:Infusions, Intravenous; D007421:Intestine, Small / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D007511:Ischemia / Q000628:therapy*; D008297:Male; D008641:Mesenteric Vascular Occlusion / Q000209:etiology; D051379:Mice; D008813:Mice, Inbred ICR; D010289:Parenteral Nutrition, Total; D009195:Peroxidase / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015427:Reperfusion Injury / Q000209:etiology / Q000378:metabolism*
D017382:Reactive Oxygen Species; D005973:Glutamine; D009195:Peroxidase