Animal, page 1 of 8 © The Animal Consortium 2019

doi:10.1017/S1751731119000223
animal

Dietary alanyl-glutamine improves growth performance of

weaned piglets through maintaining intestinal morphology and
digestion–absorption function
T. D. Zou , C. X. Denga, Z. R. Wang, Y. L. Ye and J. M. You†
a

Jiangxi Province Key Laboratory of Animal Nutrition, Engineering Research Center of Feed Development, Jiangxi Agricultural University, Nanchang 330045, Jiangxi,
China

(Received 30 December 2017; Accepted 21 January 2019)

Alanyl-glutamine (Ala-Gln), a highly soluble and stable glutamine dipeptide, is known to improve gut integrity and function.
The aim of this study was to evaluate whether dietary Ala-Gln supplementation could improve growth performance, intestinal
development and digestive-absorption function in weaned piglets. A total of 100 purebred Yorkshire piglets weaned at 21 days of
age were assigned randomly to four dietary treatment groups and fed a basal diet (control group) or a basal diet containing
0.15%, 0.30% and 0.45% Ala-Gln, respectively. Compared with the control group, piglets fed the Ala-Gln diets had higher average
daily gain and lower feed : gain and diarrhea rate (P < 0.05). Moreover, dietary Ala-Gln supplementation increased villous height
and villous height : crypt depth ratio in duodenum and jejunum (P < 0.05), as well as the activities of maltase and lysozyme in
jejunum mucosa (P < 0.05). In addition, a decrease in serum diamine oxidase activity and crypt depth in duodenum and jejunum
was observed in piglets fed the Ala-Gln diets (P < 0.05). Serum cytosolic phospholipase A2 (cPLA2) concentration and gene
expression of cPLA2, Na+-dependent glucose transporter 1, glucose transporter 2 and peptide transporter 1 in jejunum were
increased by feeding Ala-Gln diets relative to control diet (P < 0.05). These results indicated that feeding Ala-Gln diet has beneficial
effects on the growth performance of weaned piglets, which associated with maintaining intestinal morphology and digestive-
absorption function.

Keywords: alanyl-glutamine, intestinal structure, intestinal disaccharidase activities, nutrient transportation, mucosal barrier function

Implications
The gut can undergo long-lasting developmental changes
throughout one’s life. Weaning is the most severe, early
life stress for piglets and is associated with increased
susceptibility to intestinal dysfunction. It is important to
better understand how nutritional interventions at early
postnatal life stages affect intestinal structure, physiology
and function in weaned piglets. The novel finding was
that feeding alanyl-glutamine (Ala-Gln), a highly soluble
and stable glutamine dipeptide, improves growth per-
formance in weaned piglets, which associated with better
intestinal morphological and capability of digestion and
absorption. Current results may provide new insights into
developing Ala-Gln as a functional feed supplement in pig
industry.
a
These two authors contributed equally to this work.
†
E-mail: youjinm@163.com
Introduction
The gut, which plays a pivotal role in the digestion–absorption
of nutrients and regulation of the mucosal response to exo-
genous antigens, undergoes long-lasting developmental
changes throughout one’s life (Boyer et al., 2015; Miner-
williams and Moughan, 2016). The environmental factors at
early postnatal life stages which in addition to genetic factor,
including nutritional stimulus, pathogen infection and stress,
have been shown to induce significant changes in intestinal
structure, physiology and disease susceptibility in mamma-
lians (Pluske et al., 1997; O’Brien et al., 2001; Boudry et al.,
2004). Weaning is the most severe, early life stress for piglets
and is associated with increased susceptibility to intestinal
dysfunction, including increased intestinal permeability (Smith
et al., 2010), hypersecretion (Moeser et al., 2007), oxidative
injury (Yin et al., 2014), inflammation and barrier disruption
(Boudry et al., 2004; Pié et al., 2004). as well as remarkable
changes in morphology (Nabuurs et al., 1993). Thus, devel-
oping a novel feeding strategy for protecting the integrity of

1
Zou, Deng, Wang, Ye and You
intestinal structure and maintaining the function of intestinal
mucosa barrier at postweaning period is particularly important
to alleviate stress-associated growth depression and syndrome
in weaned piglets.
Glutamine (Gln), a critical gut-trophic nutrient, is a major
respiratory fuel for enterocytes and has been shown to pro-
tect against intestinal epithelial cell damage in weaned pigs
(Wu et al., 1996; Yi et al., 2005). However, a limitation of
using Gln as a dietary supplement is that it is unstable in
solution or during prolonged storage where it can hydrolyze
to the toxic byproducts (Khan and Elia, 1991; Carneiro-Filho
et al., 2003). This led to the development of more stable
synthetic forms. As a Gln dipeptide, Ala-Gln is highly soluble
in water, favorable thermal stability and well tolerated, and
commonly used in clinical practice (Jian et al., 1999) and
experimental settings (Moore et al., 2015; Zhang et al.,
2016). Furthermore, Ala-Gln performs better in terms of
blood L-glutamine availability than the free form of Gln, and
is more beneficial for the organism as a whole (Minguetti-
Câmara et al., 2014; Rosa et al., 2015). It is suggested that
supplementation with Ala-Gln can positively act ameliorat-
ing intestinal injury and malabsorption in undernourished
children (Lima et al., 2007) and murine models (Ueno et al.,
2011). Proposed the mechanisms of improving gut integrity
and epithelial cell homeostasis of Ala-Gln include increased
epithelial proliferation and the expression of tight junction
proteins (Ueno et al., 2011; Santos et al., 2013; Moore et al.,
2015) and decreased intestinal inflammation, mucosa
hyperplasia and enterocytes apoptosis (Rodrigues et al.,
2013). Nevertheless, to our knowledge, the effects of Ala-Gln
supplementation on growth performance aspects of intest-
inal development and digestion–absorption function in
weaned piglets are poorly understood.
This study aimed to investigate the effects of dietary Ala-
Gln supplementation on growth performance, intestinal
morphology and digestion–absorption function in weaned
piglets. Furthermore, our findings may provide new insights
into developing Ala-Gln as a functional feed supplement in
pig industry.
Material and methods
Experimental design, animal and diets
All procedures involving animals were approved by the Ani-
mal Care and Use Committee of Jiangxi Agricultural Uni-
versity. A total of 100 purebred Yorkshire weaned male
piglets (21 ± 2 days of age, 6.64 ± 0.20 kg) were selected
from a commercial pig unit and randomly assigned to one of
four dietary treatments (five replicate pens per treatment and
five pigs per pen). The four dietary treatments consisted of
the basal diet treatment (control group) and the basal diet
supplemented with 0.15%, 0.30% or 0.45% Ala-Gln (Sigma-
Aldrich, 99% purity, Saint Louis, MO, USA). The basal diet
was formulated to meet the National Research Council
(2012)-recommended nutrient requirements for pigs weigh-
ing 5 to 10 kg, and ingredient and nutrient composition were
2
presented in Supplementary Table S1. The experimental diet
was formulated using Ala-Gln to substitute for the same
amount of corn in the basal diet. All pigs were housed in
temperature-controlled room with free access to food and
water. The initial temperature was maintained at 28°C for
the 1st week, and then gradually decreased by 2°C per week
for the end of the experiment. The experimental period lasted
21 days. The fasted pigs were weighed individually in the
morning of days 8, 15 and 22, and feed consumption per pen
was measured daily. Average daily feed intake (ADFI), aver-
age daily gain (ADG) and feed : gain (F/G) were calculated.
Diarrhea index
The diarrhea of all pigs was observed throughout the
experimental period, and fecal consistency was scored as
follows: 0, normal; 1, pasty; 2, semiliquid; and 3, liquid. Pigs
with daily fecal consistency scores of ⩾ 2 were defined as
diarrhea (Liu et al., 2014). The diarrhea rate is expressed as
the ratio of the number of piglets with diarrhea to the total
number of piglets in each treatment group, and the cumu-
lative rate of diarrhea was calculated.
Blood sampling and analysis
On days 8, 15 and 22 of the trial after overnight fasting, one
pig with weight close to the average level per pen was
selected for bleeding via precaval vein. Blood samples were
allowed to coagulate for 30 min and centrifuged (3000 × g,
4°C and 15 min) to obtain serum. Serum diamine oxidase
(DAO) activity was determined using spectrophotometry as
previously described (Hou et al., 2012). Serum cytosolic
phospholipase A2 (cPLA2) level was measured using a
commercial RIA kit (Beijing Sinouk Institute of Biological
Technology, Beijing, China) according to the manufacturer’s
instructions.
Tissue sample collection
At the end of the experiment, one piglet with the average BW
in each pen was selected and slaughtered by intravenous
injection of with pentobarbital sodium according to protocols
approved by the Jiangxi Agricultural University Animal Care
Advisory Committee. The small intestines were isolated
quickly and sampled. Approximately 2 cm segments of duo-
denum (10 cm from the stomach), the middle jejunum and
ileum (15 cm from the cecum) were isolated, washed and
fixed in 4% paraformaldehyde for histological analysis. The
jejunal specimen obtained from the same site was fixed in
2.5% glutaraldehyde in 0.1 M phosphate buffer for ultra-
structural examination. The small intestinal mucosa samples
were collected by scraping the intestinal segments with a
microscope slide, frozen in liquid nitrogen and stored at
−80°C for enzyme activity and gene expression analysis.
Intestinal morphology analysis
The fixed intestinal segments were embedded in paraffin.
Then, 5-μm thick tissue sections were deparaffinized, rehy-
drated and stained with hematoxylin and eosin. At least four
images per section and five sections from each individual

piglet were acquired. The villus height and crypt depth of the
intestinal mucosa (duodenum, jejunum and ileum) were
measured on the images using an Olympus CK 40 microscope
(Olympus Optical Company, Tokyo, Japan) and the average
was used for the calculations. Then villi–crypt ratio (VCR) was
calculated.
Transmission electron microscopy
The glutaraldehyde fixed jejunal samples were washed, dehy-
drated and embedded in Epon-812 (Shell Chemical Co., New
York, USA). At least four ultrathin sections for each jejunal
sample were stained with 2% uranyl acetate, poststained with
0.2% lead citrate and examined with a Hitachi H-600 trans-
mission electron microscope (Hitachi, Tokyo, Japan).
Measurement of enzymes activities
The activities of disaccharidase (sucrase, lactase and mal-
tase), Na+, K+-ATPase and lysozyme in jejunal mucosa were
measured using assay kits purchased from Nanjing Jiancheng
Bioengineering Institute (Nanjing, China) according to the
manufacturer’s instructions, and expressed as on a per-
milligram protein basis. The protein concentration was
determined by Pierce BCA Protein Assay kit (Thermo Scien-
tific, Waltham, MA, USA).
Quantitative real-time PCR
Total RNA was extracted from the small intestinal mucosa by
using TRIzol reagent (Sigma, Saint Louis, MO, USA) according
to the manufacturer’s instructions. Ribonucleic acid con-
centration was determined by nucleic-acid/protein analyzer
(Beckman Coulter DU800; Beckman Coulter Inc., Fullerton,
CA, USA). The integrity of RNA was confirmed by 1% agarose
gel electrophoresis. Complementary DNA (cDNA) was syn-
thesized with the iScriptTM cDNA Synthesis Kit (Bio-Rad, Her-
cules, CA, USA). Real-time quantitative PCR was conducted on
the ABI Prism 7300 sequence detector (Applied Biosystems,
Foster City, CA, USA) as described previously with β-actin used
as a reference gene (Ipharraguerre et al., 2013). The relative
messenger RNA (mRNA) expression of analyzed genes was
calculated using the method of 2− ΔΔCt (Livak and Schmittgen,
2001). The primer sequences are listed in Supplementary
Table S2.
Statistical analysis
The pen was used as the experimental unit for growth per-
formance and diarrhea rate, while the selected pig in each pen
was considered as the experimental unit for all other variables.
We verified that the model residuals were normally distributed
using the Shapiro–Wilk test and required no transformation
before analysis. Data were analyzed using SAS (SAS Institute
Inc., Cary, NC, USA) following a randomized block design. The
preplanned contrast (0 v. others) was assessed using the GLM
procedure in SAS. The linear and quadratic responses to diet-
ary Ala-Gln concentrations were determined using the REG
procedure of SAS. A value of P < 0.05 was considered to be
statistically significant, whereas P values between 0.05 and
Alanyl-glutamine improves intestinal development
0.1 were considered as trends. All data were presented as
means with their standard errors.
Results
Growth performance
For the whole experimental period, dietary supplementation of
Ala-Gln led to a decrease in F/G ratio (P = 0.038) and an
increase in ADG (P < 0.001) compared with the control group.
Increasing the level of Ala-Gln from 0% to 0.45% increased
the ADG (linear and quadratic; P = 0.002 and P = 0.016,
respectively). However, there was no significant effect on ADFI
with increasing levels of Ala-Gln (P > 0.05) (Table 1).
As shown in Table 2, piglets fed the Ala-Gln diets had
lower (P < 0.05) diarrhea rates than those fed the control
diet during the overall period. Meanwhile, a linear decrease
in diarrhea rate was observed as Ala-Gln supplementation
increased as each week, respectively, as well as the whole
experimental period (P < 0.05).
Intestinal morphology
The data of intestinal morphology were shown in Table 3. In
the duodenum, dietary supplementation of Ala-Gln signifi-
cantly increased villous height (P = 0.003) and the VCR index
(P = 0.002) and decreased crypt depth (P = 0.009) compared
with the control group. Increasing the level of Aln-Gln from
0% to 0.45% increased villous height (linear and quadratic;
P = 0.025 and P = 0.020, respectively) and the VCR index
(linear and quadratic; P = 0.006 and P = 0.006, respectively),
and decreased crypt depth (linear and quadratic; P = 0.011
and P = 0.018, respectively). A similar histomorphalogic
changes was noticed in Jejunum. In addition, compared with
the control group, examination of the ultrastructural
morphology showed a markedly increase in microvillus
height and density in jejunum of piglets fed the Ala-Gln diets
(Figure 1). In the ileum, piglets fed the Ala-Gln diets had
higher villus height (P = 0.015) and VCR index (P = 0.025)
than those fed the control diet. Furthermore, the villus height
showed linear and quadratic dose responses to the levels of
Ala-Gln supplementation (P = 0.007 and P = 0.012, respec-
tively). A linear increase in the VCR index was also observed
as Ala-Gln supplementation increased (P = 0.025).
The activities of disaccharidase and lysozyme in
jejunal mucosa
As shown in Table 4. Piglets fed the Ala-Gln diets had
markedly increased the activities of maltase (P = 0.018) and
lysozyme (P = 0.043), and a tendency to increase sucrase
activity (P = 0.056) compared with those fed the control diet.
Furthermore, there was a significant linear increase in maltase
activity (P = 0.032) and quadratic increase in lysozyme activity
(P = 0.019) with the increasing levels of Ala-Gln.
Serum concentration and messenger RNA expression in small
intestinal mucosa of cytosolic phospholipase A2
Dietary supplementation with 0.30% or 0.45% Ala-Gln
increased serum cPLA2 concentration compared with the
control group (P < 0.05, Figure 2a). Expression level of
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Zou, Deng, Wang, Ye and You

Table 1 Effects of dietary alanyl-glutamine (Ala-Gln) supplementation on the growth performance of weaned pigs
Ala-Gln supplementation (%) P-value

0 0.15 0.30 0.45 SEM 0 v. others L Q

Initial BW (kg) 6.62 6.63 6.65 6.65 0.20 0.953 0.949 0.989
Final BW (kg) 12.91 13.63 14.03 13.78 0.27 0.151 0.238 0.390
ADG (g/day)
Days 0 to 7 202.1 238.6 273.9 262.7 10.8 0.019 0.022 0.230
Days 7 to 14 328.6 373.5 377.9 367.2 7.6 0.008 0.065 0.057
Days 14 to 21 355.7 374.6 391.4 377.6 6.9 0.114 0.199 0.251
Days 0 to 21 295.4 328.9 347.7 335.8 5.9 < 0.001 0.002 0.016
ADFI (g/day)
Days 0 to 7 285.6 304.2 322.6 328.2 12.9 0.285 0.242 0.811
Days 7 to 14 525.0 534.8 541.3 529.9 12.1 0.721 0.859 0.689
Days 14 to 21 635.5 642.1 652.7 651.7 13.5 0.682 0.658 0.899
Days 0 to 21 482.0 493.7 505.5 503.3 11.2 0.480 0.489 0.774
F/G ratio
Days 0 to 7 1.42 1.29 1.18 1.24 0.04 0.029 0.049 0.177
Days 7 to 14 1.60 1.43 1.43 1.45 0.03 0.018 0.096 0.128
Days 14 to 21 1.81 1.72 1.66 1.73 0.05 0.386 0.603 0.410
Days 0 to 21 1.61 1.48 1.43 1.49 0.03 0.038 0.114 0.145
ADG = average daily gain; ADFI = average daily feed intake; F/G = feed : gain; L = linear; Q = quadratic.
n = 5 for each group.

Table 2 Effect of dietary alanyl-glutamine (Ala-Gln) supplementation on the diarrhea rate (%) of weaned pigs
Ala-Gln supplementation (%) P-value

0 0.15 0.30 0.45 SEM 0 v. others L Q

Days 0 to 7 26.00 21.67 21.11 15.33 1.75 0.063 0.030 0.826

Days 7 to 14 23.21 17.85 12.14 10.71 1.64 0.012 0.003 0.466
Days 14 to 21 18.33 6.67 3.33 2.50 1.79 < 0.001 < 0.001 0.032
Days 0 to 21 22.91 15.20 10.73 9.52 1.43 < 0.001 < 0.001 0.073
L = linear; Q = quadratic.
n = 5 for each group.

Table 3 Effects of dietary alanyl-glutamine (Ala-Gln) supplementation on intestinal morphology of weaned pigs
Ala-Gln supplementation (%) P-value

0 0.15 0.30 0.45 SEM 0 v. others L Q

Villus height (μm)

Duodenum 298.09 321.60 329.66 319.46 3.95 0.003 0.025 0.020
Jejunum 292.04 314.95 337.53 329.81 6.17 0.020 0.023 0.229
Ileum 236.28 245.82 272.20 251.76 3.74 0.015 0.007 0.012
Crypt depth (μm)
Duodenum 182.46 172.79 157.53 169.86 2.72 0.009 0.011 0.018
Jejunum 215.50 152.07 148.32 175.73 6.65 < 0.001 0.002 < 0.001
Ileum 166.93 159.07 146.15 148.62 4.14 0.102 0.074 0.528
VCR
Duodenum 1.63 1.87 2.10 1.89 0.05 0.002 0.006 0.006
Jejunum 1.36 2.07 2.32 1.87 0.08 < 0.001 < 0.001 < 0.001
Ileum 1.43 1.58 1.90 1.69 0.06 0.025 0.020 0.090
VCR = villus height : crypt depth ratio; L = linear; Q = quadratic.
n = 5 for each group.

mucosal cPLA2 mRNA in the duodenum, jejunum and ileum (P < 0.05) compared with those fed the control diet
of pigs fed 0.15% or 0.30% Ala-Gln diet were higher (Figure 2b).

4
 Alanyl-glutamine improves intestinal development

Figure 1 Effects of dietary alanyl-glutamine (Ala-Gln) supplementation on the jejunal ultrastructure of weaned pigs. (a) the basal diet; (b) basal diet and
supplementation with 0.15% Ala-Gln; (c) basal diet and supplementation with 0.30% Ala-Gln; (d) basal diet and supplementation with 0.45% Ala-Gln.

Table 4 Effects of dietary alanyl-glutamine (Ala-Gln) supplementation on the activities of disaccharidase and lysozyme in jejunal mucosa of
weaned pigs
Ala-Gln supplementation (%) P-value

U/mg protein 0 0.15 0.30 0.45 SEM 0 v. others L Q

Lactase 55.93 61.45 64.29 58.86 2.52 0.350 0.628 0.314

Maltase 90.02 114.25 131.31 123.31 6.23 0.018 0.032 0.168
Sucrase 82.69 93.99 102.81 95.84 3.40 0.056 0.115 0.177
Na +/K+-ATPase 0.137 0.150 0.158 0.152 0.008 0.408 0.519 0.582
Lysozyme 71.52 88.99 101.31 81.38 4.14 0.043 0.212 0.019
L = linear; Q = quadratic.
n = 5 for each group.

Figure 2 Effects of dietary alanyl-glutamine (Ala-Gln) supplementation on serum concentration (a) and messenger RNA (mRNA) expression in small
intestinal mucosa (b) of cytosolic phospholipase A2 (cPLA2) in weaned pigs. Data are expressed as means ± SEM, n = 5. *P < 0.05.

Serum diamine oxidase activity Discussion

As shown in Figure 3, dietary supplementation with 0.30%
It has been well established that early life environmental
Ala-Gln significantly decreased serum DAO activity com-
factors, such as diets and stress, have a remarkable impact
pared with other groups at days 14 and 21 (P < 0.05).
on long-term intestinal health and disease susceptibility (Pié
et al., 2004; Yi et al., 2005). Alanyl-glutamine, a stable Gln
Gene expression of nutrient transporters in small intestinal
dipeptide, shown to performed better in terms of blood Gln
mucosa
availability than the free form (Minguetti-Câmara et al.,
As shown in Figure 4, dietary supplementation of Ala-Gln
2014). Several studies have shown that Ala-Gln could pro-
increased the mRNA expression levels of Na+-dependent
mote intestinal integrity and epithelial cell homeostasis
glucose transporter 1 (SGLT1) and peptide transporter 1
in vitro and in animal models (Ueno et al., 2011; Zhou et al.,
(PEPT1) in the jejunum compared with control group
2012; Rodrigues et al., 2013). The present study was con-
(P < 0.05). Expression level of glucose transporter 2 (GLUT2)
ducted to evaluate whether dietary Ala-Gln supplementation
mRNA in the jejunum of piglets fed 0.30% or 0.45% Ala-Gln
could improve growth performance, intestinal development
diet was higher than those fed the control diet (P < 0.05).

5
Zou, Deng, Wang, Ye and You
and digestion–absorption function using pig as animal
model. Here, we observed that Ala-Gln supplementation was
beneficial to growth performance with less occurrence of
diarrhea in piglets, which are similar to previous report
demonstrating that intravenous administration of Ala-Gln
had a positive effect on gain performance and the occurrence
of diarrhea in early weaned calves (Zhou et al., 2012). In
order to clarify the mechanism of growth-promoting effect by
supplementing Ala-Gln, the intestinal responses of weaned
piglets to Ala-Gln diet were further investigated.
The high value of the VCR ratio has been considered as a
useful indicator of a high digestion and absorption capacity
of the small intestine (Montagne et al., 2003). In this study,
Figure 3 Effects of dietary alanyl-glutamine (Ala-Gln) supplementation
on serum diamine oxidase (DAO) activity of weaned pigs. Data are
expressed as means ±SEM, n = 5. *P < 0.05.
Figure 4 Effects of dietary alanyl-glutamine (Ala-Gln) supplementation on mucosal messenger RNA (mRNA) expression of nutrient transporters in the
duodenum, jejunum and ileum of weaned pigs. (a to c) mRNA expression of Na+-dependent glucose transporter 1 (SGLT1) (a), glucose transporter 2
(GLUT2) (b) and peptide transporter 1 (PEPT1) (c). Data are expressed as means ±SEM, n = 5. *P <0.05.
6
feeding Ala-Gln diets increased the ratio of VCR in duode-
num, jejunum and ileum of weaned piglets, which are an
important factor to promote the postnatal growth. Further-
more, our results showed that alteration of intestinal mor-
phology, to some degree, displayed the dose response to the
levels of Ala-Gln supplementation. Consistent with the
positive influence of Ala-Gln on intestinal morphology, an
increased maltase activity in the jejunum was observed in
Ala-Gln-supplemented piglets. In addition, it has been
demonstrated that lysozyme exerted beneficial effect on
nutrient digestibility, intestinal morphology and barrier, as
well as gut microflora in weaned piglets (Oliver and Wells,
2013; Ma et al., 2017). Correspondingly, dietary Ala-Gln
supplementation markedly increased the lysozyme activity in
jejunum of weaned piglets. Similar with previous reports in
juvenile red drum, Sciaenops ocellatus, which found that
addition of Gln increased serum lysozyme activity (Cheng,
2011).
To determine whether Ala-Gln could regulate intestinal
absorption function in piglets, we examined the gene
expression of nutrient transporters in the small intestine of
piglets. The expression of SGLT1 and GLUT2 play an impor-
tant role in the absorption and uptake of glucose in small
intestine. The SGLT1 mediates intestinal glucose absorption
and utilization and was crucial to the glucose-induced
incretion hormone secretion (Röder et al., 2014). Glucose
transporter 2 transported glucose and fructose from enter-
ocytes into the systemic system (Zhang et al., 2016). It has
been demonstrated that early weaning stress significantly

decreased the expression of SGLT1 mRNA in jejunum of pigs
(Zhang et al., 2006). In contrast, we found that dietary Ala-
Gln supplementation upregulated the expression of SGLT1
and GLUT2 in jejunum of weaned piglets. In addition, studies
have shown that the transfer and absorption of oligopeptides
were closely correlated with the PEPT1 gene expression
(Buyse et al., 2001). In this research, the jejunum PEPT1
mRNA level was significantly increased in Ala-Gln group.
Together, these results provide evidence that feeding Ala-Gln
diets could increase the capacity of nutrients digestion and
absorption, thus resulting in the better feed conversion ration
relative to piglets fed control diet.
In addition, cPLA2 is the rate-limiting enzyme for the gen-
eration of prostaglandins, which are bioactive lipids that have
been proved to be important in maintaining gastrointestinal
homeostasis (Dey et al., 2006; Panel et al., 2006). Our results
showed that dietary Ala-Gln supplementation increased serum
cPLA2 concentration and the expression of cPLA2 mRNA in
duodenum and jejunum of weaned piglets. As previously
reported, serum DAO activity may be a useful biomarker for
evaluating intestinal mucosal injury and disease (Kamei et al.,
2005). The clinic study showed that enteral supplementation
Gln caused a significant decrease in plasma DAO activity and
abated the degree of intestine injury in severe burned patients
(Xi et al., 2004). In this study, there was decreasing serum
DAO activity in Ala-Gln-supplemented piglets relative to con-
trol piglets. Besides, serum DAO decreased with the lower
supplementation of Ala-Gln but increased with the higher
supplementation, which implied that the alterations of serum
DAO activity did not presented the dose response to the levels
of Ala-Gln supplementation. Taken together, these data indi-
cate that Ala-Gln had an effect on maintaining intestinal
integrity and improving the mucosal barrier function in
weaned piglets, which may explain the reason why Ala-Gln
had beneficial effect on the growth performance.
In conclusion, we have demonstrated that feeding Ala-Gln
has beneficial effects in alleviating growth depression in
weaned piglets, mainly associated with better intestinal mor-
phological function and capability of digestion and absorption.
Supplementary material
To view supplementary material for this article, please visit
https://doi.org/10.1017/S1751731119000223
Acknowledgment
This study was supported by the National Natural Science
Foundation of China (31360554) and the Scientific Research
Fund of Jiangxi Province Education Department (GJJ12216). The
authors are grateful to Prof. Min Du for reviewing and editing
the manuscript.
Declaration of interest
The authors declare no conflicts of interest.
Alanyl-glutamine improves intestinal development
Ethics statement
All animal experimental and care procedures followed Experi-
mental Animal Care and Use Guidelines of China and were
approved by the Animal Care and Use Committee of Jiangxi
Agricultural University.
Software and data repository resources
The authors declare that the data of this study are not deposited
in any official repository.
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