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Dulaglutide markedly prevents peritoneal fibrosis in a rodent model of chronic kidney disease: Insights into the pathogenesis

  • Authors:
    • Chih-Chao Yang
    • Ya Yue
    • Yi-Ting Wang
    • John Y. Chiang
    • Ben-Chung Cheng
    • Tsuen-Wei Hsu
    • Yi-Ling Chen
    • Yi-Chen Li
    • Hon-Kan Yip
  • View Affiliations / Copyright

    Affiliations: Division of Nephrology, Department of Internal Medicine, Kaohsiung Chang Gung Memorial Hospital and College of Medicine, Chang Gung University, Kaohsiung 83301, Taiwan, R.O.C., Institute of Nephrology and Blood Purification, The First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, Guangdong 510632, P.R. China, Institute for Translational Research in Biomedicine, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung 83301, Taiwan, R.O.C., Department of Computer Science and Engineering, National Sun Yat-Sen University, Kaohsiung 804201, Taiwan, R.O.C., Clinical Medicine Research Center, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan 701401, Taiwan, R.O.C.
    Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 151
    |
    Published online on: July 21, 2025
       https://doi.org/10.3892/ijmm.2025.5592
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Abstract

Peritoneal fibrosis (PF) is a major complication of long-term peritoneal dialysis, leading to ultrafiltration failure and technique dropout, highlighting the urgent need for therapies that can preserve peritoneal membrane function and longevity. The present study evaluated the effectiveness of dulaglutide in preserving the functional integrity and durability of the peritoneum while inhibiting PF. In vitro Met-5A cells showed significant upregulation of inflammatory, oxidative stress, intracellular and mitochondrial reactive oxygen species (ROS), fibrotic, intracellular cytoskeletal, apoptotic and epithelial-mesenchymal transition (EMT) biomarkers, and dipeptidyl peptidase 4 (DPP4), following stimulation with a uremic toxin (p-Cresol), PF inducer [chlorhexidine gluconate (CG)] or endotoxin [lipopolysaccharide (LPS)]. Notably, these effects were significantly suppressed by dulaglutide or TGF-β/DPP4 double silencing. Furthermore, cell viability and glucagon-like peptide 1 (GLP-1) expression displayed an opposite pattern to ROS among the groups. Sprague-Dawley rats were divided into the following groups: i) Sham control (SC); ii) chronic kidney disease (CKD); iii) CKD + CG (mimicking renal failure and PF); and iv) CKD + CG + dulaglutide, and were euthanized by day 42. At this time point, the highest levels of peritoneal protein expression levels of oxidative stress (NOX-1, NOX-2 and DPP4), inflammation (NF-κB and TNF-α), angiogenesis (CD31 and von Willebrand factor) and EMT (TGF-β, Snail, β-catenin, vimentin, phosphorylated-Smad3, α-smooth muscle actin, collagen I, N-cadherin and fibronectin) factors; and cellular expression levels of fibrosis and inflammation markers, were observed in the CKD + CG group, the lowest were detected in the SC group, and the levels were significantly reduced in the CKD + CG + dulaglutide group compared with those in the CKD group. Furthermore, the expression levels of antioxidant proteins (nuclear factor erythroid 2-related factor 2, NAD(P)H quinone oxidoreductase 1 and GLP-1 receptor) exhibited an opposite trend to ROS-associated proteins among the groups. Additional Sprague-Dawley rats were categorized into the following groups: i) SC; ii) LPS-induced peritonitis; iii) LPS-induced peritonitis + dulaglutide, and were euthanized by day 5 after peritonitis induction. At this time point, flow cytometry revealed significantly increased levels of inflammatory cells (CD11b/c+, myeloperoxidase+ and Ly6G+ cells) in the circulation and abdominal fluid, and increased peritoneal permeability in the LPS-induced peritonitis group compared with those in the SC group; these levels were significantly reversed in the LPS-induced peritonitis + dulaglutide group. In conclusion, dulaglutide may effectively maintain peritoneal integrity primarily by suppressing inflammation, oxidative stress, EMT and fibrosis.
View Figures

Figure 1

Serial changes of serum levels of
BUN, creatinine, DPP4 and GLP-1 and RUp/c. Circulating
levels of (A) BUN, (B) creatinine, (C) DPP4 and (D) GLP-1, and (E)
RUp/c at baseline. Circulating levels of (F) BUN, (G)
creatinine, (H) DPP4 and (I) GLP-1, and (J) RUp/c at day
14 after CKD induction. Circulating levels of (K) BUN, (L)
creatinine, (M) DPP4 and (N) GLP-1, and (O) RUp/c at day
35 after CKD induction. Circulating levels of (P) BUN, (Q)
creatinine, (R) DPP4 and (S) GLP-1, and (T) RUp/c at day
42 after CKD induction. All statistical analyses were performed by
one-way ANOVA, followed by Bonferroni's multiple comparisons post
hoc test (n=6/group). #P<0.05, ##P<0.01
vs. SC; *P<0.05 vs. CKD; ns, not significant. BUN,
blood urea nitrogen; CG, chlorhexidine gluconate; CKD, chronic
kidney disease; DPP4, dipeptidyl peptidase 4; GLP-1, glucagon-like
peptide 1; RuP/uC, ratio of urine protein/urine
creatinine; SC, sham control.

Figure 2

Protein expression levels of
epithelial-mesenchymal transistion biomarkers in the peritonium by
day 42 after CKD induction. (A) Protein expression levels were
detected by western blot analysis. Semi-quantification of (B)
TGF-β, (C) Snail, (D) β-catenin, (E) p-Smad3/Smad3, (F) vimentin,
(G) α-SMA, (H) collagen I, (I) N-cadherin and (J) fibronectin. All
statistical analyses were performed by one-way ANOVA, followed by
Bonferroni's multiple comparisons post hoc test (n=6/group).
#P<0.05, ##P<0.01 vs. SC;
*P<0.05 vs. CKD. α-SMA, α-smooth actin, CKD, chronic
kidney disease; CG, chlorhexidine gluconate; p-, phosphorylated;
SC, sham control.

Figure 3

Protein expression levels of
biomarkers of oxidative stress, inflammation and angiogenesis, as
well as DPP4, GLP-1R and antioxidants in the peritonium by day 42
after CKD induction. (A) Protein expression levels were detected by
western blot analysis. Semi-quantification of (B) NOX-1, (C) vWF,
(D) GLP-1R, (E) NOX-2, (F) TNF-α, (G) Nrf2, (H) NQO-1, (I)
p-NF-κB/NF-κB, (J) CD31, (K) DPP4 and (L) VEGF. All statistical
analyses were performed by one-way ANOVA, followed by Bonferroni's
multiple comparisons post hoc test (n=6/group).
#P<0.05, ##P<0.01 vs. SC;
*P<0.05 vs. CKD. CG, chlorhexidine gluconate; CKD,
chronic kidney disease; DPP4, dipeptidyl peptidase 4; GLP-1R,
glucagon-like peptide 1 receptor; NQO-1, NAD(P)H-quinone
oxidoreduc-tase 1; Nrf2, nuclear factor erythroid 2-related factor
2; p-, phosphorylated; SC, sham control; vWF, von Willebrand
factor.

Figure 4

Histopathological findings in the rat
peritonium by day 42 after CKD induction. Light microsope findings
(magnification, ×400) of hematoxylin and eosin staining for the
detection of thickness and hyperplasia (double yellow arrows), as
well as the number of small vessels and muscularized vessels (red
arrows) in the intimmal layer of the peritoneum. Scale bar, 20
µm. Histological images of the (A) SC group, (B) CKD group,
(C) CKD + CG group and (D) CKD + CG + dulaglutide group. (E)
Thickness of the parietal layer of the peritoneum. (F) Hyperplasia
area of parietal layer of peritoneum. (G) Number of small vessels
(defined as ≤25 µm). (H) Thickness of muscularized vessels;
defined as vessels exhibiting medial layer hyperplasia resulting in
thickness being increased >50% of internal lumen diameter. All
statistical analyses were performed by one-way ANOVA, followed by
Bonferroni's multiple comparisons post hoc test (n=6/group).
#P<0.05, ##P<0.01 vs. SC;
*P<0.05 vs. CKD. CG, chlorhexidine gluconate; CKD,
chronic kidney disease; SC, sham control. Figure 4. Histopathological findings in
the rat peritonium by day 42 after CKD induction. Light microsope
findings (magnification, ×400) of hematoxylin and eosin staining
for the detection of thickness and hyperplasia (double yellow
arrows), as well as the number of small vessels and muscularized
vessels (red arrows) in the intimmal layer of the peritoneum. Scale
bar, 20 µm. Histological images of the (A) SC group, (B) CKD
group, (C) CKD + CG group and (D) CKD + CG + dulaglutide group. (E)
Thickness of the parietal layer of the peritoneum. (F) Hyperplasia
area of parietal layer of peritoneum. (G) Number of small vessels
(defined as ≤25 µm). (H) Thickness of muscularized vessels;
defined as vessels exhibiting medial layer hyperplasia resulting in
thickness being increased >50% of internal lumen diameter. All
statistical analyses were performed by one-way ANOVA, followed by
Bonferroni's multiple comparisons post hoc test (n=6/group).
#P<0.05, ##P<0.01 vs. SC;
*P<0.05 vs. CKD. CG, chlorhexidine gluconate; CKD,
chronic kidney disease; SC, sham control.

Figure 5

Fibrosis and inflamamotry cell
infiltration in the rat peritoneum by day 42 after CKD induction.
Microscopic findings (magnification, ×400) of Masson's trichrome
staining, which was used to identify the fibrotic area (blue), in
the (A) SC group, (B) CKD group, (C) CKD + CG group and (D) CKD +
CG + dulaglutide group. (E) Percentage of fibrotic area.
Microscopic findings (magnification, ×400) of immunohistochemistry,
which was performed to detect the expression levels of MMP-9
(green), in the (F) SC group, (G) CKD group, (H) CKD + CG group and
(I) CKD + CG + dulaglutide group. (J) Number of positively stained
MMP-9 cells. Scale bar, 20 µm. All statistical analyses were
performed by one-way ANOVA, followed by Bonferroni's multiple
comparisons post hoc test (n=6/group). #P<0.05,
##P<0.01 vs. SC; *P<0.05 vs. CKD. CG,
chlorhexidine gluconate; CKD, chronic kidney disease; SC, sham
control.

Figure 6

Quantitative evaluation of kidney
injury score and inflammatory cell infiltration in the kidney
parenchyma by day 42 after CKD induction. Microscopic examination
(magnification, ×200x; hematoxylin and eosin staining)
demonstrating an increased loss of brush border in renal tubules
(yellow arrows), tubular necrosis (green arrows), tubular
dilatation (red asterisk), protein cast formation (black asterisk)
and dilatation of Bowman's capsule (blue arrows) in the CKD group
than in the other groups. Scale bar, 50 µm. Images from the
(A) SC group, (B) CKD group, (C) CKD + CG group and (D) CKD + CG +
dulaglutide group. (E) Kidney injury scores. Microscopic findings
(magnification, ×400) of immunofluorescence staining, which was
performed to assess the cellualr expression of MMP-9 (green). Scale
bar, 20 µm. Images of the (F) SC group, (G) CKD group, (H)
CKD + CG group and (I) CKD + CG + dulaglutide group. (J) Number of
MMP-9+ cells. Statistical analyses were performed by (E)
Kruskal-Wallis's test and Dunn's post hoc test or (J) one-way
ANOVA, followed by Bonferroni's multiple comparisons post hoc test
(n=6/group). #P<0.05, ##P<0.01 vs. SC;
*P<0.05 vs. CKD. CG, chlorhexidine gluconate; CKD,
chronic kidney disease; PF, peritoneal fibrosis; SC, sham
control.

Figure 7

LPS induces peritoneal damaged by day
5 of treatment. Circulating levels of (A) GLP-1 and (B) DPP4 at
baseline. Circulating levels of (C) GLP-1 and (D) DPP4 at day 5.
Abdominal levels of (E) GLP-1 and (F) DPP4 at day 5. Circulating
levels of the indicator of peritoneal permeability FITC-dextran (G)
10, (H) 20 and (I) 30 min after LPS treatment. Flow cytometric
analysis of the number of (J) CD11b/c+, (K)
MPO+ anf (L) Ly6G+ cells in circulation. Flow
cytometric analysis of the number of (M) CD11b/c+, (N)
MPO+ cells and (O) Ly6G+ cells in the
abdominal fluid. All statistical analyses were performed by one-way
ANOVA, followed by Bonferroni's multiple comparisons post hoc test
(n=6/group). #P<0.05 vs. SC; *P<0.05
vs. LPS. DPP4, dipeptidyl peptidase 4; GLP-1, glucagon-like peptide
1; LPS, lipopolysaccharide; MPO, myeloperoxidase; SC, sham
control.

Figure 8

Identification of histopathological
changes in the mesothelial layer of the peritoneum by day 5 of LPS
treatment. Light microscopic examination (magnification, ×100) of
peritoneal hyperplasia and thickness (white) by hematoxylin and
eosin staining in the (A) SC group, (B) LPS group and (C) LPS +
dulaglutide group. Scale bars, 100 µm. Increased
magnification (magnification, ×400) of images in SC, LPS and LPS +
dulaglutide groups. Scale bars, 20 µm. Condensed collagen
fiber deposition (pink color) and neovascularization (green arrows)
were notably increased in LPS group and more substantially
increased in LPS + dulaglutide group than in SC group. (D) Levels
of peritoneal hyperplasia. (E) Thickness of the mesothelial layer
in the peritoneum. Microscopic findings (magnification, ×200) of
Masson's trichrome staining, which was used to identify peritoneal
fibrosis (blue color), in the (F) SC group, (G) LPS group and (H)
LPS + dulaglutide group. (I) Fibrotic area. All statistical
analyses were performed by one-way ANOVA, followed by Bonferroni's
multiple comparisons post hoc test (n=6/group).
#P<0.05 vs. SC; *P<0.05 vs. LPS. LPS,
lipopolysaccharide; SC, sham control.

Figure 9

Schematic diagram of the proposed
underlying mechanims of dulaglutide treatment of peritoneal
fibrosis and PD failure. CKD, chronic kidney disease; DPP4,
dipeptidyl peptidase 4; EMT, epithelial-mesenchymal transion;
GLP-1, glucagon-like peptide 1; GLP-1R, GLP-1 receptor; LPS,
lipopolysac-charide; NQO-1, NAD(P)H-quinone oxidoreductase 1; Nrf2,
nuclear factor erythroid 2-related factor 2; PD, pertoneal
dialysis.
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Copy and paste a formatted citation
Spandidos Publications style
Yang C, Yue Y, Wang Y, Chiang JY, Cheng B, Hsu T, Chen Y, Li Y and Yip H: Dulaglutide markedly prevents peritoneal fibrosis in a rodent model of chronic kidney disease: Insights into the pathogenesis. Int J Mol Med 56: 151, 2025.
APA
Yang, C., Yue, Y., Wang, Y., Chiang, J.Y., Cheng, B., Hsu, T. ... Yip, H. (2025). Dulaglutide markedly prevents peritoneal fibrosis in a rodent model of chronic kidney disease: Insights into the pathogenesis. International Journal of Molecular Medicine, 56, 151. https://doi.org/10.3892/ijmm.2025.5592
MLA
Yang, C., Yue, Y., Wang, Y., Chiang, J. Y., Cheng, B., Hsu, T., Chen, Y., Li, Y., Yip, H."Dulaglutide markedly prevents peritoneal fibrosis in a rodent model of chronic kidney disease: Insights into the pathogenesis". International Journal of Molecular Medicine 56.4 (2025): 151.
Chicago
Yang, C., Yue, Y., Wang, Y., Chiang, J. Y., Cheng, B., Hsu, T., Chen, Y., Li, Y., Yip, H."Dulaglutide markedly prevents peritoneal fibrosis in a rodent model of chronic kidney disease: Insights into the pathogenesis". International Journal of Molecular Medicine 56, no. 4 (2025): 151. https://doi.org/10.3892/ijmm.2025.5592
Copy and paste a formatted citation
x
Spandidos Publications style
Yang C, Yue Y, Wang Y, Chiang JY, Cheng B, Hsu T, Chen Y, Li Y and Yip H: Dulaglutide markedly prevents peritoneal fibrosis in a rodent model of chronic kidney disease: Insights into the pathogenesis. Int J Mol Med 56: 151, 2025.
APA
Yang, C., Yue, Y., Wang, Y., Chiang, J.Y., Cheng, B., Hsu, T. ... Yip, H. (2025). Dulaglutide markedly prevents peritoneal fibrosis in a rodent model of chronic kidney disease: Insights into the pathogenesis. International Journal of Molecular Medicine, 56, 151. https://doi.org/10.3892/ijmm.2025.5592
MLA
Yang, C., Yue, Y., Wang, Y., Chiang, J. Y., Cheng, B., Hsu, T., Chen, Y., Li, Y., Yip, H."Dulaglutide markedly prevents peritoneal fibrosis in a rodent model of chronic kidney disease: Insights into the pathogenesis". International Journal of Molecular Medicine 56.4 (2025): 151.
Chicago
Yang, C., Yue, Y., Wang, Y., Chiang, J. Y., Cheng, B., Hsu, T., Chen, Y., Li, Y., Yip, H."Dulaglutide markedly prevents peritoneal fibrosis in a rodent model of chronic kidney disease: Insights into the pathogenesis". International Journal of Molecular Medicine 56, no. 4 (2025): 151. https://doi.org/10.3892/ijmm.2025.5592
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