Abstract
Introduction: Alanyl-glutamine (Ala-Gln) is a compound known for its protective effects in various tissue injuries. However, its role in asthma-related lung injuries remains underexplored. This study investigates the mechanisms by which Ala-Gln modulates sDPP4-induced airway epithelial-mesenchymal transition and ovalbumin (OVA)-induced asthma in a mouse model. Methods: An asthma model was established in female C57BL/6 J mice by using OVA. CD4+ T cells and bronchial epithelial cells (BECs) were isolated from the spleen and bronchi of the mice, respectively. Interventions included recombinant sCD26/sDPP4 protein, Ala-Gln, and EX527 (a SIRT1 inhibitor). Flow cytometry was used to assess Th17 and Treg cell populations. Mice were treated with Ala-Gln, EX527, and budesonide (BUD). Histopathological changes in lung tissues were evaluated using hematoxylin-eosin and Masson staining. White blood cell counts were measured with a hematology analyzer. The expression levels of DPP4, IL-17, SIRT1, SMAD2/3, N-cadherin, E-cadherin, MMP9, and α-SMA proteins were analyzed. Results: Treatment with recombinant sCD26/sDPP4 resulted in decreased E-cadherin expression in BECs and increased levels of α-SMA, MMP9, and N-cadherin, effects that were mitigated by Ala-Gln. Ala-Gln also prevented the reduction in SIRT1 expression in BECs and the increase in Th17 cell differentiation induced by recombinant sCD26/sDPP4. EX527 administration alongside Ala-Gln reversed these changes and enhanced the phosphorylation of SMAD2/3 through SIRT1 signaling. BUD alone reduced inflammation and fibrosis in bronchial tissue and lowered the Th17/Treg ratio in peribronchial lymph nodes. The therapeutic effect of BUD was further improved with concurrent Ala-Gln treatment. Conclusion: Ala-Gln can inhibit BEC fibrosis and Th17 cell differentiation mediated by recombinant sCD26/sDPP4 through the SIRT1 pathway. Combined with BUD, Ala-Gln enhanced therapeutic efficacy in OVA-induced asthma in mice, which could offer improved outcomes for asthmatic patients with elevated DPP4 levels.
Journal Section:
Experimental Immunology – Research Article
References
1.
Wan
R
, Srikaram
P
, Guntupalli
V
, Hu
C
, Chen
Q
, Gao
P
. Cellular senescence in asthma: from pathogenesis to therapeutic challenges
. EBioMedicine
. 2023
;94
:104717
. 2.
Kim
J
, Ham
J
, Kang
HR
, Bae
YS
, Kim
T
, Kim
HY
. JAK3 inhibitor suppresses multipotent ILC2s and attenuates steroid-resistant asthma
. Sci Adv
. 2023
;9
(51
):eadi3770
. 3.
Alcorn
JF
, Crowe
CR
, Kolls
JK
. TH17 cells in asthma and COPD
. Annu Rev Physiol
. 2010
;72
:495
–516
. 4.
Banuelos
J
, Cao
Y
, Shin
SC
, Lu
NZ
. Immunopathology alters Th17 cell glucocorticoid sensitivity
. Allergy
. 2017
;72
(3
):331
–41
. 5.
Ghorpade
DS
, Ozcan
L
, Zheng
Z
, Nicoloro
SM
, Shen
Y
, Chen
E
, et al. Hepatocyte-secreted DPP4 in obesity promotes adipose inflammation and insulin resistance
. Nature
. 2018
;555
(7698
):673
–7
. 6.
Li
L
, Ling
F
, Li
R
, Jiang
Y
, Long
H
, Xiao
B
, et al. DPP4 regulates the Th17/IL-17 Axis and accelerates epithelial mesenchymal transition to promote ovalbumin-induced asthma in female C57bl/6J mice
. Front Biosci
. 2023
;28
(12
):342
. 7.
Anderson
ED
, Alishahedani
ME
, Myles
IA
. Epithelial-mesenchymal transition in atopy: a mini-review
. Front Allergy
. 2020
;1
:628381
. 8.
Han
H
, Chung
SI
, Park
HJ
, Oh
EY
, Kim
SR
, Park
KH
, et al. Obesity-induced vitamin D deficiency contributes to lung fibrosis and airway hyperresponsiveness
. Am J Respir Cell Mol Biol
. 2021
;64
(3
):357
–67
. 9.
Zhang
S
, Fan
Y
, Qin
L
, Fang
X
, Zhang
C
, Yue
J
, et al. IL-1β augments TGF-β inducing epithelial-mesenchymal transition of epithelial cells and associates with poor pulmonary function improvement in neutrophilic asthmatics
. Respir Res
. 2021
;22
(1
):216
. 10.
Mao
Z
, Qian
Y
, Liu
Z
, Shi
Y
, Fan
L
, Zhang
Q
. LINC00158 modulates the function of BEAS-2B cells via targeting BCL11B and ameliorates OVA-LPS-induced severe asthma in mice models
. Int Immunopharmacol
. 2024
;130
:111739
. 11.
Yang
Y
, Liu
Y
, Wang
Y
, Chao
Y
, Zhang
J
, Jia
Y
, et al. Regulation of SIRT1 and its roles in inflammation
. Front Immunol
. 2022
;13
:831168
. 12.
Shen
P
, Deng
X
, Chen
Z
, Ba
X
, Qin
K
, Huang
Y
, et al. SIRT1: a potential therapeutic target in autoimmune diseases
. Front Immunol
. 2021
;12
:779177
. 13.
Jiang
YZ
, Huang
XR
, Chang
J
, Zhou
Y
, Huang
XT
. SIRT1: an intermediator of key pathways regulating pulmonary diseases
. Lab Invest
. 2024
;104
(5
):102044
. 14.
Chen
Z
, Xie
W
, Tang
S
, Lin
M
, Ren
L
, Huang
X
, et al. Taraxerone exerts antipulmonary fibrosis effect through Smad signaling pathway and antioxidant stress response in a Sirtuin1-dependent manner
. Phytother Res
. 2024
;38
(7
):3720
–35
. 15.
Xia
Y
, Luo
Q
, Gao
Q
, Huang
C
, Chen
P
, Zou
Y
, et al. SIRT1 activation ameliorates rhesus monkey liver fibrosis by inhibiting the TGF-β/smad signaling pathway
. Chem Biol Interact
. 2024
;394
:110979
. 16.
Fukuda
Y
, Akimoto
K
, Homma
T
, Baker
JR
, Ito
K
, Barnes
PJ
, et al. Virus-induced asthma exacerbations: SIRT1 targeted approach
. J Clin Med
. 2020
;9
(8
):2623
. 17.
Wang
Y
, Le
Y
, Wu
J
, Zhao
W
, Zhang
Q
, Xu
G
, et al. Inhibition of xanthine oxidase by allopurinol suppresses HMGB1 secretion and ameliorates experimental asthma
. Redox Biol
. 2024
;70
:103021
. 18.
Ferrantelli
E
, Liappas
G
, Vila Cuenca
M
, Keuning
ED
, Foster
TL
, Vervloet
MG
, et al. The dipeptide alanyl-glutamine ameliorates peritoneal fibrosis and attenuates IL-17 dependent pathways during peritoneal dialysis
. Kidney Int
. 2016
;89
(3
):625
–35
. 19.
Liu
SK
, Ma
LB
, Yuan
Y
, Ji
XY
, Sun
WJ
, Duan
JX
, et al. Alanylglutamine relieved asthma symptoms by regulating gut microbiota and the derived metabolites in mice
. Oxid Med Cell Longev
. 2020
;2020
:7101407
. 20.
Hou
YC
, Pai
MH
, Liu
JJ
, Yeh
SL
. Alanyl-glutamine resolves lipopolysaccharide-induced lung injury in mice by modulating the polarization of regulatory T cells and T helper 17 cells
. J Nutr Biochem
. 2013
;24
(9
):1555
–63
. 21.
Hou
YC
, Liu
JJ
, Pai
MH
, Tsou
SS
, Yeh
SL
. Alanyl-glutamine administration suppresses Th17 and reduces inflammatory reaction in dextran sulfate sodium-induced acute colitis
. Int Immunopharmacol
. 2013
;17
(1
):1
–8
. 22.
McMillan
SJ
, Lloyd
CM
. Prolonged allergen challenge in mice leads to persistent airway remodelling
. Clin Exp Allergy
. 2004
;34
(3
):497
–507
. 23.
Liu
ZH
, Zhang
Y
, Wang
X
, Fan
XF
, Zhang
Y
, Li
X
, et al. SIRT1 activation attenuates cardiac fibrosis by endothelial-to-mesenchymal transition
. Biomed Pharmacother
. 2019
;118
:109227
. 24.
Li
HT
, Chen
ZG
, Lin
YS
, Liu
H
, Ye
J
, Zou
XL
, et al. CpG-ODNs and budesonide act synergistically to improve allergic responses in combined allergic rhinitis and asthma syndrome induced by chronic exposure to ovalbumin by modulating the TSLP-DC-ox40l Axis
. Inflammation
. 2018
;41
(4
):1304
–20
. 25.
Braga-Neto
MB
, Oliveira
BM
, Rodrigues
RS
, Noronha
FJ
, Leitao
RF
, Brito
GA
, et al. Protective effects of alanyl-glutamine supplementation against nelfinavir-induced epithelial impairment in IEC-6 cells and in mouse intestinal mucosa
. Cancer Biol Ther
. 2012
;13
(14
):1482
–90
. 26.
Sun
J
, Chu
S
, Lu
M
, Pan
Q
, Li
D
, Zheng
S
, et al. The roles of dipeptidyl peptidase-4 and its inhibitors in the regulation of airway epithelial-mesenchymal transition
. Exp Lung Res
. 2020
;46
(6
):163
–73
. 27.
Chen
Z
, Yu
J
, Fu
M
, Dong
R
, Yang
Y
, Luo
J
, et al. Dipeptidyl peptidase-4 inhibition improves endothelial senescence by activating AMPK/SIRT1/Nrf2 signaling pathway
. Biochem Pharmacol
. 2020
;177
:113951
. 28.
Liu
Y
, Shi
G
. Roles of sirtuins in asthma
. Respir Res
. 2022
;23
(1
):251
. 29.
Guan
R
, Wang
J
, Cai
Z
, Li
Z
, Wang
L
, Li
Y
, et al. Hydrogen sulfide attenuates cigarette smoke-induced airway remodeling by upregulating SIRT1 signaling pathway
. Redox Biol
. 2020
;28
:101356
. 30.
Cruzat
VF
, Keane
KN
, Scheinpflug
AL
, Cordeiro
R
, Soares
MJ
, Newsholme
P
. Alanyl-glutamine improves pancreatic β-cell function following ex vivo inflammatory challenge
. J Endocrinol
. 2015
;224
(3
):261
–71
. 31.
Zhang
L
, Li
L
, Zhou
M
, Zhou
QY
, Tang
JH
, Liang
M
, et al. Association of serum YKL-40 and DPP4 with T2-high asthma in Chinese adults
. Medicine
. 2024
;103
(6
):e37169
. 32.
Okaya
T
, Kawasaki
T
, Sato
S
, Koyanagi
Y
, Tatsumi
K
, Hatano
R
, et al. Functional roles of CD26/DPP4 in bleomycin-induced pulmonary hypertension associated with interstitial lung disease
. Int J Mol Sci
. 2024
;25
(2
):748
. 33.
Liu
C
, Xu
J
, Fan
J
, Liu
C
, Xie
W
, Kong
H
. DPP-4 exacerbates LPS-induced endothelial cells inflammation via integrin-α5β1/FAK/AKT signaling
. Exp Cell Res
. 2024
;435
(1
):113909
. 34.
Sato
S
, Kawasaki
T
, Hatano
R
, Koyanagi
Y
, Takahashi
Y
, Ohnuma
K
, et al. Functional roles of CD26/DPP4 in lipopolysaccharide-induced lung injury
. Am J Physiol Lung Cell Mol Physiol
. 2024
;326
(5
):L562
–73
. 35.
Deinhardt-Emmer
S
, Deshpande
S
, Kitazawa
K
, Herman
AB
, Bons
J
, Rose
JP
, et al. Role of the senescence-associated factor dipeptidyl peptidase 4 in the pathogenesis of SARS-CoV-2 infection
. Aging Dis
. 2024
;15
(3
):1398
–415
. 36.
Liu
J
, Zong
C
, Yu
X
, Ding
Y
, Chang
B
, Wang
R
, et al, Ala-Gln) Ameliorates Dextran Sulfate Sodium (DSS)-Induced Acute Colitis by Regulating the Gut Microbiota
. Alanyl-glutamine (Ala-Gln) ameliorates dextran sulfate sodium (DSS)-Induced acute colitis by regulating the gut microbiota, PI3K-akt/NF-κB/STAT3 signaling, and associated pulmonary injury
. ACS Infect Dis
. 2023
;9
(4
):979
–92
. 37.
Huang
L
, Hough
O
, Vellanki
RN
, Takahashi
M
, Zhu
Z
, Xiang
YY
, et al. L-alanyl-L-glutamine modified perfusate improves human lung cell functions and extend porcine ex vivo lung perfusion
. J Heart Lung Transpl
. 2023
;42
(2
):183
–95
. 38.
Shang
Q
, Zhu
L
, Shang
W
, Zeng
J
, Qi
Y
. Dioscin exhibits protective effects on in vivo and in vitro asthma models via suppressing TGF-β1/Smad2/3 and AKT pathways
. J Biochem Mol Toxicol
. 2022
;36
(8
):e23084
. 39.
Ruteaga-Navarro
TC
, Reyes-Romero
MA
, Torres-Salazar
QL
. Effect of sitagliptin on down-regulation of KAT7 and SIRT1 gene expression in breast cancer cell line MCF7
. Cir Cir
. 2023
;91
(3
):334
–8
. 40.
Sun
J
, Hou
Z
, Xia
T
, Liu
C
, Huang
Q
, Wei
L
. The effect of budesonide-formoterol powder inhalation on cough and lung function following thoracoscopic pulmonary surgery: a retrospective cohort study
. Interdiscip Cardiovasc Thorac Surg
. 2024
:ivae097
. 41.
Tessier
B
, Moine
L
, Peramo
A
, Tsapis
N
, Fattal
E
. Poly(malic acid)-budesonide nanoconjugates embedded in microparticles for lung administration
. Drug Deliv Transl Res
. 2024
;14
(8
):2062
–78
. 42.
Türkeli
A
, Yilmaz
Ö
, Karaman
M
, Kanik
ET
, Firinci
F
, İnan
S
, et al. Anti-VEGF treatment suppresses remodeling factors and restores epithelial barrier function through the E-cadherin/β-catenin signaling axis in experimental asthma models
. Exp Ther Med
. 2021
;22
(1
):689
. 43.
Hillman
NH
, Kemp
MW
, Royse
E
, Grzych
H
, Usada
H
, Ikeda
H
, et al. Postnatal budesonide improved lung function in preterm lambs exposed to antenatal steroids and chorioamnionitis
. Pediatr Res
. 2024
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