Journal of Surgical Research 140, 77– 83 (2007)

doi:10.1016/j.jss.2006.10.021

Glutamine Donor Pretreatment in Rat Kidney Transplants with Severe

Preservation Reperfusion Injury 1
T. Florian Fuller, M.D.,*,2 Florian Rose, M.D.,† Kristen D. Singleton, M.S.,‡ Yvonne Linde, B.S.,†
Uwe Hoff, M.D.,† Chris E. Freise, M.D.,§ Duska Dragun, Ph.D.,† and Claus U. Niemann, M.D.¶
*Department of Urology and †Department of Nephrology, Charité University Hospital CCM, Berlin, Germany; ‡Department of
Anesthesiology, University of Colorado Health Sciences Center, Denver, Colorado; §Department of Surgery, Division of Transplantation
and ¶Department of Anesthesia and Perioperative Care, University of California San Francisco, San Francisco, California

Submitted for publication August 8, 2006

Conclusions. In rat renal grafts suffering severe PRI

Background. Glutamine (GLN) has been shown to
confer cytoprotection by enhancing endogenous heat
shock protein (HSP) expression. We hypothesized
that GLN donor pretreatment protects rat renal
grafts against severe preservation reperfusion in-
jury (PRI).
Materials and methods. GLN (0.75 g/kg) or saline was
administered i.p. to male donor rats 24 h and 6 h before
donor nephrectomy. Kidneys (n ⴝ 6/group) were cold-
stored in UW solution for 40 h and transplanted
into bilaterally nephrectomized syngeneic recipients.
Grafts were removed after 24 h. Renal HSP 70 expres-
sion was determined by Western blotting. Graft func-
tion was assessed by serum creatinine. Renal cross
sections were microscopically examined for acute tu-
bular necrosis, apoptosis, tubular proliferation, and
macrophage infiltration.
Results. GLN donor pretreatment significantly in-
creased intragraft HSP 70 expression. Serum creati-
nine was not different between groups: 2.6 ⴞ 0.2 mg/dL
(saline) versus 2.7 ⴞ 0.5 mg/dL (GLN). Both treatment
groups showed severe tubular damage with signifi-
cantly less papillary necrosis in the GLN group (P <
0.05). GLN significantly reduced the number of apopto-
tic tubular cells in the cortex, medulla, and papilla
(P < 0.001 versus saline). Postinjury tubular prolifera-
tion, measured by PCNA antigen expression, and in-
tragraft macrophage infiltration was not influenced
by GLN.
1
This work was presented at the World Transplant Congress, July
2006, Boston, Massachusetts.
2
To whom correspondence and reprint requests should be ad-
dressed at Klinik fuer Urologie, Charité Universitaetsmedizin Ber-
lin, Campus Charité Mitte, Schumannstrasse 20/21, 10117 Berlin,
Germany. E-mail: Florian.Fuller@charite.de.
pharmacological preconditioning with GLN attenu-
ates early structural damage, especially tubular cell
apoptosis. Stimulation of renal HSP 70 expression
could be an important mechanism of GLN-induced cy-
toprotection. Our findings may have implications for
the treatment of delayed graft function in recipients of
marginal donor kidneys. © 2007 Elsevier Inc. All rights reserved.
Key Words: kidney transplantation; rat; ischemic in-
jury; glutamine; heat shock protein; apoptosis.
INTRODUCTION
Kidneys from marginal donors have an increased
incidence of delayed graft function (DGF). Clinically
applicable donor pretreatment protocols to treat DGF
in recipients of marginal renal grafts are not well es-
tablished. We used a syngeneic rat kidney transplan-
tation model with severe preservation reperfusion in-
jury (PRI), induced by 40 h of cold storage, to test
potential benefits of glutamine donor pretreatment.
Glutamine (GLN), an amino acid conditionally es-
sential during critical illness and injury, has been
shown to reduce cell and organ damage induced by
endotoxemia or ischemia [1, 2]. As previously shown,
glutamine induces endogenous heat shock protein 70
(HSP 70) expression in animals and humans, thus
conferring cytoprotection against various stressors
[3, 4]. Glutamine has been reported to improve out-
comes in experimental models of intestinal and cardiac
ischemia reperfusion injury (IRI) [2, 5]. In animal mod-
els of renal IRI, elevated HSP 70 levels have been
associated with improved functional outcomes [6, 7].
In our present study, we hypothesized that glu-
tamine donor pretreatment, initiated 24 h before organ

77 0022-4804/07 $32.00
© 2007 Elsevier Inc. All rights reserved.
78 JOURNAL OF SURGICAL RESEARCH: VOL. 140, NO. 1, JUNE 1, 2007
procurement, induces renal HSP 70 expression and
attenuates early structural damage and functional im-
pairment in 40 h cold-preserved syngeneic rat kidney
grafts.
MATERIALS AND METHODS
GLN Administration and Experimental Design
GLN was administered as an alanyl-glutamine dipeptide
(Fresenius-Kabi, Homburg, Germany), which was dissolved in sa-
line. GLN solutions were filtered with a 0.45-␮m filter before intra-
peritoneal administration. Donor rats received either 0.75 g/kg GLN
or saline (n ⫽ 6 per group) 24 h and 6 h before left donor nephrec-
tomy. Grafts were cold-stored in UW solution for 40 h and trans-
planted into bilaterally nephrectomized syngeneic recipients. In the
sham operation group, animals (n ⫽ 3) underwent right nephrec-
tomy and the contralateral kidney was used for additional experi-
ments. Animals were sacrificed 24 h after surgery for blood sampling
and organ harvesting. Kidneys were cross-sectioned and stored in
preparation for Western blot analysis, conventional histology, and
immunohistochemistry.
Animal Surgery
All animal protocols were reviewed and approved by the Univer-
sity of California San Francisco Committee on Animal Research, and
animal care was in agreement with the National Institutes of Health
(NIH) guidelines for ethical animal research (NIH publication num-
ber 80-123, revised in 1985). Kidney donors and recipients were
inbred male Lewis rats (200 to 250 g; Charles River Laboratories,
Wilmington, MA) housed under standard conditions with free access
to water and chow. All procedures were performed under inhalation
anesthesia with isoflurane. Left kidneys were procured, flushed with
ViaSpan, University of Wisconsin (UW) solution and stored at 4°C
for 40 h.
Recipients underwent bilateral native nephrectomies followed
by heterotopic kidney transplantation using an established micro-
surgical technique [8]. Briefly, end-to-side anastomoses between
the renal vessels and the recipient’s abdominal aorta and inferior
vena cava were created using continuous 8-0 nylon sutures; mean
warm ischemia time was 17 ⫾ 1.2 min. An end-to-end uretero-
ureterostomy was performed using interrupted 11-0 nylon suture.
After recovery from anesthesia, animals were transferred to the
housing facility and monitored until sacrifice 24 h post-trans-
plantation.
Heat Shock Protein Detection
Renal cross sections including cortex, medulla, and papilla were
snap frozen in liquid nitrogen and stored at ⫺80°C until analysis.
Western blotting was performed as previously described [1]. Blots
were blocked with a 5% milk PBS Blotto solution. For HSP 70
detection, blots were incubated with a primary mouse-anti-HSP 70
antibody (Stressgen, Victoria, Canada). Blots were washed and in-
cubated with a horseradish peroxidase-conjugated secondary goat-
anti-mouse antibody (Santa Cruz Biotechnology, Santa Cruz, CA).
Densitometry was determined using the UVP Chemiluminescent
Darkroom System (UVP Inc., Upland, CA).
Graft Function
Graft function was assessed by serum creatinine measurement
24 h post-transplantation. Recipient blood samples (0.5 mL) were
processed by IDDEX Veterinary Services (Sacramento, CA).
Tubular Necrosis Score
For morphological studies, formalin-fixed renal grafts (n ⫽ 6 per
group) were used. Paraffin cross sections (4 ␮m) were stained with
hematoxylin and eosin and periodic acid-Schiff (PAS) using standard
procedures. A renal pathologist, blinded for the experimental condi-
tions, examined and scored kidney sections for acute tubular necrosis
(ATN) as previously described [9]. ATN was graded as follows: 0
represented no abnormalities, and 1, 2, 3, and 4 represented slight
(⬍20%), moderate (20 to 40%), severe (40 to 60%), and near-total
(⬎60%) necrosis of the renal parenchyma, respectively.
Apoptosis Detection
Renal apoptosis was examined using the in situ Cell Death De-
tection Kit (Roche Diagnostics, Indianapolis, IN) based on the TdT-
mediated dUTP nick end labeling (TUNEL) method. Paraffin-
embedded renal cross sections including the cortex, medulla, and
papilla were treated according to the manufacturer’s instructions
and stained with Fluorescein. TUNEL-positive apoptotic cells were
visualized under a fluorescence microscope (Zeiss Axio Imager A1,
Jena, Germany) and quantitated using a digital imaging system
(Zeiss Axiocam HR with Axiovision 4.4 software). In each microcom-
partment, 10 to 15 randomly chosen fields of view (FOV) per section
were evaluated. The means of six samples were grouped together to
obtain a final mean ⫾ SD. Positive cell staining was recorded digi-
tally and expressed as the percentage of TUNEL-positive area per
FOV at 400⫻ magnification.
Immunohistochemistry
For assessment of intragraft monocyte/macrophage infiltration a
monoclonal mouse anti-ED1 antibody (endothelial 1 antigen on
monocytes/macrophages; Serotec, Oxford, United Kingdom) was
used. For assessment of tubular cell proliferation a monoclonal
mouse anti-PCNA (proliferating cell nuclear antigen, clone PC10;
Zymed Laboratories Inc., San Francisco, CA) was used. Immunohis-
tochemistry was carried out on 2 ␮m paraffin sections. ED1⫹ stain-
ing was performed using the alkaline-phosphatase-anti-alkaline-
phosphatase (APAAP) method on paraffin sections after depar-
affinization and rehydration. Sections were incubated for 60 min at
room temperature with primary Ab at 1:1000 dilution in RPMI
(Seromed, Heidelberg, Germany) with 10% fetal calf serum (FCS)
and 3% bovine serum albumin (BSA). Fast Red Kit (DakoCytoma-
tion, Hamburg, Germany) was used for detection and visualization.
PCNA staining was performed using the standard avidin-biotin-
complex method (Dako). Sections were deparaffinized in xylol, rehy-
drated through graded alcohols and cooked in citrate buffer (pH 6.0)
for 5 min. Slides were then incubated with primary Ab (diluted at
1:100) for 60 min at room temperature in Ab-diluent (Dako). AEC-
chromogen (Dako) was used for visualization. Sections incubated
with corresponding isotype controls instead of primary Ab were used
as negative controls.
Intragraft ED1 and PCNA expression were evaluated in a blinded
fashion. In the cortex, medulla, and papilla, 10 to 15 randomly
chosen FOVs per section were evaluated using a light microscope
(Zeiss Axio Imager A1, Jena, Germany). Positive cell staining was
expressed as mean ⫾ SD of cells per field of view (FOV).
Statistical Analysis
Values are expressed as means ⫾ SD. If not otherwise indicated,
groups were compared with the unpaired Student’s t-test. For com-
 FULLER ET AL.: GLUTAMINE IN RAT KIDNEY TRANSPLANTS
parison of graded variables (i.e., tubular necrosis score) the nonpara-
metric Mann-Whitney U test was used. All tests were two-tailed. A
P value of ⬍0.05 was considered to be significant. All statistical
analyses were performed with the statistical package SPSS for Win-
dows (Version 10.07; SPSS Inc., Chicago).
RESULTS
Renal HSP 70 Expression
Compared with saline, GLN pretreatment signifi-
cantly increased HSP 70 expression in renal grafts
suffering severe preservation reperfusion injury (P ⬍
0.05; Fig. 1). In sham-operated controls (n ⫽ 3) renal
HSP 70 expression was low. In this group, GLN treat-
ment had no impact on renal HSP 70 expression 24 h
after surgery.
Graft Function
In the sham group (right nephrectomy) mean serum
creatinine on postoperative day 1 was 0.5 ⫾ 0.1 mg/dL.
Graft function was not different between saline and GLN
pretreatment groups. The following serum-creatinine
levels were measured 1 d post transplantation in recipi-
ents of 40 h cold-stored grafts: 2.6 ⫾ 0.2 mg/dL (saline)
versus 2.7 ⫾ 0.5 mg/dL (GLN; P ⫽ not significant).
Tubular Necrosis Score
In sham-operated animals, no tubular necrosis was
seen. Grafts transplanted after 40 h of cold storage
showed severe tubular damage in the cortex and me-
dulla, whereas the papilla showed moderate to severe
damage (Fig. 2). Compared with saline, GLN donor
pretreatment significantly reduced papillary necrosis
FIG. 1. Renal HSP 70 expression 24 h after transplantation and
70 h after the last GLN dose (0.75 g/kg). Tissue homogenates for
Western blot analysis were obtained from isograft cross-sections
containing cortex, medulla, and papilla. Blot represents one of five
experiments with similar results. GLN pretreatment significantly
increased renal HSP 70 expression relative to saline donor pretreat-
ment (*P ⬍ 0.05; Student’s U test).
79
FIG. 2. Acute tubular necrosis (ATN) score 1 d post kidney
transplantation (KTX). Tubular damage was graded on a scale from
0 to 4 (0 ⫽ no abnormalities; 4 ⫽ near total necrosis). Significant
differences between saline and GLN donor pretreatment were seen
in the papilla of 40 h cold-stored rat kidney grafts (*P ⬍ 0.05;
Mann-Whitney U test).
(P ⬍ 0.05; Figs. 2 and 4). In the cortex and medulla of
GLN pretreated kidneys, a trend toward less tubular
necrosis was seen. However, differences between treat-
ment groups were not statistically significant (Fig. 2).
Tubular Cell Apoptosis
Apoptotic tubular epithelial cells were detected us-
ing the TUNEL-method and visualized under a fluo-
rescence microscope (Fig. 5). Sham-operated animals
showed only a small number of TUNEL-positive apo-
ptotic cells. Forty h of cold storage and transplantation
induced marked tubular cell apoptosis in the cortex,
medulla, and papilla (Figs. 3 and 5). Compared with
saline, GLN pretreatment significantly reduced the
number of apoptotic cells in the cortex, medulla, and
papilla (P ⬍ 0.001; Fig. 3).
FIG. 3. TUNEL-positive apoptotic cells in 40 h cold-stored rat
kidney grafts 1 d post kidney transplantation (KTX). Using a fluo-
rescence microscope, positive cell staining was recorded digitally and
expressed as the percentage of TUNEL-positive area per field of
view. GLN pretreatment 24 h before donor nephrectomy and trans-
plantation significantly reduced the number of apoptotic cells in the
cortex, medulla and papilla (**P ⬍ 0.001; saline versus GLN; Stu-
dent’s t-test).
80 JOURNAL OF SURGICAL RESEARCH: VOL. 140, NO. 1, JUNE 1, 2007

FIG. 4. Histology of the renal papilla 1 d post transplantation. (A) and (B) show representative cross-sections of papillary collecting ducts
(ducts of Bellini) in 40 h cold-stored rat kidney grafts. Saline pretreated grafts show large areas of complete papillary necrosis (A), while GLN
pretreated grafts show dilated ducts with predominantly vital epithelium (B). Magnification ⫻ 400.

Tubular Cell Proliferation
Tubular cell proliferation was quantified by counting
PCNA-positive cells. In sham-operated animals, only a
few scattered PCNA-positive cells were detected. Forty
h of cold storage and transplantation induced marked
proliferative activity in the cortex and medulla, while
no PCNA-positive cells could be detected in the papilla
of either group (Table 1). GLN donor pretreatment had
no influence on proliferation of tubular epithelial cells
recovering from severe PRI (Table 1).
Macrophage Infiltration
One day after severe PRI, intragraft macrophage
infiltration was evaluated by counting the number of
ED1-positive cells per field of view (Table 2). No ED-1
positive cells were seen in native kidneys after sham
operation. In the cortex and medulla of 40 h cold-stored
grafts, ED1-positive cell counts were higher than in the
papilla. Overall, GLN pretreatment had no effect on
intragraft macrophage accumulation (Table 2).
DISCUSSION
To our knowledge, this is the first report on GLN-
induced renoprotection in rat kidney transplants with
severe ischemic injury caused by prolonged cold preser-
vation. We found a strong antiapoptotic effect associated
with GLN donor pretreatment. This effect was equally
 FULLER ET AL.: GLUTAMINE IN RAT KIDNEY TRANSPLANTS 81

FIG. 5. Fluorescein-stained apoptotic cells in representative cross-sections of 40 h cold-stored rat kidneys 1 d post-transplantation. Areas
of confluent fluorescence in saline pretreated grafts (A), (C), (E) indicate the presence of large numbers of TUNEL-positive apoptotic cells,
while GLN pretreated grafts (B), (D), (F) show little fluorescence. Quantification of TUNEL-positive cells, using a digital imaging system
revealed a strong antiapoptotic effect of GLN donor pretreatment in all three microcompartments (see Fig. 3). Magnification ⫻ 200.

seen in the cortex, medulla, and papilla of kidney grafts
subjected to severe PRI. Ischemically damaged kidney
grafts pretreated with GLN showed significantly ele-
vated HSP 70 levels almost 3 d after the last GLN dose.
TABLE 1
Average Number of Intragraft PCNAⴙ Cells 1 Day
Post-Transplantation*
Most studies investigating renoprotective effects of
HSPs use warm ischemia models with clamping of the
renal pedicle [10, 11]. In solid organ transplantation,
however, the amount of structural damage and func-
TABLE 2
Average Number of Intragraft ED1ⴙ Infiltrates 1 Day
Post-Transplantation*

Saline (N ⫽ 6) GLN (N ⫽ 6) Saline (N ⫽ 6) GLN (N ⫽ 6)

Cortex 15.7 ⫾ 3.8 15.9 ⫾ 1.9 Cortex 3.7 ⫾ 0.8 3.8 ⫾ 0.5
Medulla 13.3 ⫾ 3.9 15.8 ⫾ 2.3 Medulla 3.6 ⫾ 0.7 3.4 ⫾ 0.6
Papilla 0 0 Papilla 1.4 ⫾ 0.3 1.8 ⫾ 0.9

P ⫽ n.s. between saline and glutamine treatment groups. P ⫽ n.s. between saline and glutamine treatment groups.
* Expressed as the number of PCNA (proliferating cell nuclear * Expressed as the number of ED1-positive cells (monocytes/
antigen)-positive cells per field of view at 400x magnification. macrophages) per field of view at 400x magnification.
82 JOURNAL OF SURGICAL RESEARCH: VOL. 140, NO. 1, JUNE 1, 2007
tional outcomes strongly correlates with the duration of
cold ischemia. We used a life supporting syngeneic rat
renal transplant model with bilateral native nephrec-
tomy of the recipient. The choice of 40 h of cold preserva-
tion was based on an earlier study involving rat kidney
transplants, where we showed that identical experimen-
tal conditions generated 1 wk recipient survival rates of
50% [12]. In rat renal transplants with prolonged cold
storage, strong inflammatory responses and irreversible
structural damage have been shown to occur early after
reperfusion [9]. We therefore tested if elevated HSP 70
levels provide renoprotection during the first 24 h post-
transplantation, probably the most critical phase of renal
PRI.
Heat shock proteins (HSPs) are a highly conserved
family of essential proteins and confer cellular protection
against various forms of cellular injury, including
ischemia-reperfusion, lung injury, and shock [3]. Short-
term hyperthermia is known as one of the most potent
inducers of endogenous HSPs. In a rat renal transplant
model, enhanced HSP 72 expression, induced by donor
hyperthermia, has been shown to protect against PRI [6].
However, in the clinical setting, short courses of hyper-
thermia as a means of HSP induction are impractical. In
recent years, several compounds, some with estab-
lished clinical applications, have been identified as in-
ducers of renal HSP 70 expression [10, 11, 13]. For
efficient pharmacological HSP 70 induction, however,
some of these drugs require relatively high dosing.
Therefore, their clinical safety remains a major con-
cern. GLN, a conditionally essential amino acid, has
been shown to efficiently induce HSP 70 expression in
vitro and in vivo [2, 5]. In both animals and humans,
GLN decreases end-organ injury and enhances sur-
vival in settings of sublethal stress [3]. Previous clini-
cal trials indicate that even at high doses, GLN has no
measurable side effects in healthy volunteers and crit-
ically ill patients [14, 15]. Our choice of GLN dosage
and route of administration was based upon previous
dose-response studies by Wischmeyer et al. [1], show-
ing that a single infusion of 0.75 g/kg GLN rapidly
increased GLN plasma levels and induced HSP 72
expression in heart and lung tissue in rats. The same
authors also showed that a single intraperitoneal dose
of 0.52 g/kg GLN, given 18 h before cardiac ischemia-
reperfusion injury, decreased postischemic myocardial
dysfunction [5].
In our present kidney transplant model, 40 h of
cold storage induced severe tubular necrosis (ATN)
and apoptosis, 24 h post-transplantation. GLN donor
pretreatment significantly reduced papillary necrosis
(Figs. 2 and 4), and slightly improved the ATN score in
the cortex and medulla (Fig. 2). Quantification of
PCNA-positive cells revealed marked proliferative ac-
tivity of proximal tubules in the cortex and medulla
(Table 1), despite severe PRI. GLN donor pretreatment
did not dampen proximal tubular cell regeneration af-
ter severe ischemic injury.
Apoptosis of tubular epithelial cells is a common
feature of ischemic and nonischemic renal injury and
typically occurs at an earlier time point than tubular
cell necrosis [16]. Over the past decade, therapeutic
strategies to interrupt the cell death cascade have been
the subject of intensive investigation. We herein dem-
onstrate a strong antiapoptotic effect of GLN, when
given to donor kidneys before cold ischemic injury and
transplantation (Figs. 3 and 5). In vitro, limited extra-
cellular GLN supplies are thought to modulate stress
and apoptotic responses [17]. Whether beneficial effects
of glutamine are purely HSP-dependent is still under
investigation. Recent experimental data, however, sug-
gest that on the molecular level, GLN-mediated cytopro-
tection strongly depends on the cellular capacity to acti-
vate an HSP response [18].
In a recent study using a rat model of hepatic warm
IRI, pharmacological preconditioning with GLN failed
to reduce intrahepatic neutrophil accumulation and did
not improve functional outcomes [19]. Previously, Suzuki
et al. showed that geranylgeranyl-acetone (GGA), an an-
tiulcer drug, ameliorates acute renal failure via HSP 70
induction in a rat model of mild IRI [11]. Pharmacological
preconditioning with GGA significantly improved renal
function and reduced macrophage infiltration after warm
IRI, generated by 30 min of renal pedicle clamping. Evi-
dence was provided that beneficial effects of GGA were
HSP 70-dependent. Using a rat renal transplant model
with 40 h of cold ischemia, Redaelli et al. [6] showed
that hyperthermia pretreatment markedly increased
renal HSP 72 expression 8 h after transplantation.
Hyperthermia pretreated grafts had a significant sur-
vival benefit and showed quicker functional recovery
within the first week post-transplantation, compared
with controls. These data suggest that induction of
renal HSP expression may help to reduce postischemic
renal dysfunction.
In the present study, donor pretreatment with GLN
did not influence the number of intragraft monocyte/-
macrophage infiltrates (Table 2). Furthermore, reno-
protective effects of GLN, as evidenced by reduced tu-
bular necrosis and apoptosis, did not translate into
improved graft function. Serum-creatinine levels of 2.7
mg/dL versus 1.1 mg/dL (Suzuki et al.) on postoperative
day 1, clearly indicate substantial differences in the
degree of renal injury, caused by either 40 h of cold
storage and transplantation or 30 min of renal pedi-
cle clamping [11]. The relatively short postoperative
follow-up of 24 h is a limitation of our present study.
Possible functional improvements, likely related to
increased intragraft HSP 70 expression, could have
become apparent with longer observation periods.
However, critical events of PRI, decisively influenc-
 FULLER ET AL.: GLUTAMINE IN RAT KIDNEY TRANSPLANTS
ing long term graft function, typically occur within
the first 24 h after transplantation [20, 21]. It is a
known fact that extensive postischemic renal tubular
injury results in early graft dysfunction. In a recent
study using a comparable experimental setup, we
showed that recipients of 39 h cold-stored rat kidney
grafts with severe papillary necrosis, induced by my-
cophenolate mofetil treatment, had a 1 wk survival
rate of 20%. In contrast, 1 wk survival in recipients of
untreated grafts with moderate papillary necrosis was
90% [22]. Therefore, GLN-induced attenuation of post-
ischemic structural damage, especially in the renal
papilla (Figs. 2 and 3), may result in improved early
graft function. In human kidney transplantation, the
use of marginal donor kidneys is often associated with
delayed graft function (DGF), resulting in poorer long-
term outcomes [21]. Since our present renal transplant
model uses “marginal” donor kidneys with severe PRI,
it may be suited to develop strategies for the treatment
of DGF.
Pharmacological HSP 70 induction in the donor kid-
ney before severe ischemic injury, combined with HSP
70 stimulation in the recipient early after transplan-
tation, may confer renoprotection and thus help to
reduce the incidence of DGF. The fact that GLN in-
duces durable HSP 70 expression, coupled with its
proven benefits and safety in clinical trials involving
critically ill patients, makes this compound ideal for
the treatment of DGF.
CONCLUSIONS
Administration of GLN a few hours before donor
nephrectomy protects rat renal grafts with severe PRI
from subsequent structural damage, in particular tu-
bular cell apoptosis. In the presence of prolonged cold
storage, HSP 70 induction could be an important mech-
anism of GLN-mediated renoprotection. Our findings
may have implications for the treatment of delayed
graft function in recipients of marginal donor kid-
neys.
ACKNOWLEDGMENTS
The authors thank Paul Wischmeyer, M.D., Department of Anes-
thesiology, University of Colorado, for providing glutamine.
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