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DKK1/SMDT1 participate in Ang II‑induced mitochondrial injury of human smooth muscle cells in an abdominal aortic aneurysm cell model

  • Authors:
    • Xiaoxia Chang
    • Baohong Yao
    • Zheng Liu
    • Zijie Guo
    • Pengyue Qiao
    • Xiaokun Li
    • Lin Wang
    • Aiqun Li
  • View Affiliations / Copyright

    Affiliations: Department of Cardiology, Muping Hospital of Traditional Chinese Medicine, Yantai, Shandong 264100, P.R. China, Department of Cardiac Surgery, Yantai Affiliated Hospital of Binzhou Medical University, Yantai, Shandong 264100, P.R. China, Department of Emergency Surgery, Yantai Affiliated Hospital of Binzhou Medical University, Yantai, Shandong 264100, P.R. China, Department of Introduction to Medicine, School of Basic Medical Sciences, Binzhou Medical University, Yantai, Shandong 264100, P.R. China, Department of Cardiology, Central Hospital Affiliated to Shandong First Medical University, Jinan, Shandong 250000, P.R. China
    Copyright: © Chang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 117
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    Published online on: February 25, 2026
       https://doi.org/10.3892/etm.2026.13112
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Abstract

Abdominal aortic aneurysm (AAA) is a lethal vascular disease, in which apoptosis and inflammation participate. However, the regulatory factors associated with aortic status remain unclear; therefore, a deeper understanding of the cell types and signaling pathways involved in the pathological process of AAAs is key to the development of medical treatments. The aim of the present study was to explore the role of dickkopf‑1 (DKK1) in angiotensin (Ang) II‑stimulated human smooth muscle cells (HSMCs), and to clarify the potential downstream targets and related pathological mechanisms of DKK1. HSMCs were stimulated with Ang II (1 µM) for 24 h; consequently, DKK1 protein expression was upregulated by 4.26‑fold and increased mRNA expression levels of IL‑1β, GRP‑94 and GRP‑78 were also observed, indicating mitochondrial and endoplasmic reticulum stress. DKK1 silencing via small interfering RNA reduced its expression, which was validated by reverse transcription‑quantitative PCR. RNA sequencing was performed in four groups: Negative control (NC), siDKK1, NC‑Ang II and siDKK1‑Ang II. Differentially expressed genes (DEGs) were identified by DESeq2, and 1,332 co‑expressed DEGs were revealed by Venn analysis, enriched in pathways, including ‘protein processing in the endoplasmic reticulum’. Robust rank aggregation prioritized key genes, notably single‑pass membrane protein with aspartate rich tail 1 (SMDT1), a mitochondrial calcium regulator. The mRNA of SMDT1 was upregulated by Ang II stimulation by 2.78‑fold, which was reversed by DKK1 silencing by 0.37‑fold. Functional enrichment associated DEGs to viral response, extracellular matrix and cytokine pathways. Protein‑protein interaction networks highlighted gene clusters via STRING. In conclusion, the present study investigated the role of DKK1 in Ang II‑induced mitochondrial dysfunction in HSMCs. To the best of our knowledge, the present study demonstrated, for the first time, that DKK1/SMDT1 mediated Ang II‑induced mitochondrial injury in HSMCs, suggesting this pathway as a potential mechanism. Therefore, the results indicated that DKK1 could serve as a therapeutic target for the prevention of AAA.

View Figures

Figure 1

Transfection efficiency of siDKK1.
(A) Reverse transcription-quantitative PCR results showed the mRNA
expression level of DKK1 in siDKK1 and NC; (B) Reverse
transcription-quantitative PCR results showed the mRNA expression
level of DKK1 in each group. *P<0.05,
***P<0.001 and ****P<0.0001;
#P<0.05 and ####P<0.0001. NC, negative
control; siDKK1, small interfering RNA for DKK1; DKK1, Dickkopf-1;
Ang II, angiotensin II.

Figure 2

Identification of downstream key
genes after DKK1 silencing. (A) Venn diagram showed co-expressed
DEGs. (B) Functional enrichment analysis of GO and KEGG of
co-expressing DEGs. (C) The top 20 genes in RRA analysis. (D) Heat
map of RRA analysis. (E) Functional enrichment analysis of GO and
KEGG of the top 20 genes in RRA analysis. (F) Reverse
transcription-quantitative PCR results showed the mRNA expression
level of SMDT1. ****P<0.0001;
####P<0.0001. NC, treated with scrambled siRNAs as
negative control; siDKK1, treated with siRNAs targeting human DKK1;
Ang II, treated with Ang II (1 µM). GO, gene ontology; KEGG, Kyoto
Encyclopedia of Genes and Genomes; NC, negative control; siDKK1,
small interfering RNA for DKK1; DKK1, Dickkopf-1; Ang II,
angiotensin II; BP, biological processes; CC, cellular components;
MF, molecular functions; DEGS, differentially expressed genes; RRA,
robust rank aggregation; SMDT1, single-pass membrane protein with
aspartate rich tail 1.

Figure 3

Ang II promotes DKK1 expression of
HSMCs and mitochondrial damage. HSMCs treated with Ang II (1 µM) or
without (control). (A) Western blotting revealed the DKK1 protein
expression levels of HSMCs. (B) Reverse transcription-quantitative
PCR results showed the mRNA expression level of IL-1β increases in
HSMCs treated with Ang II. (C) Reverse transcription-quantitative
PCR results showed the mRNA level of GRP-94 increases in HSMCs
treated with Ang II. (D) Reverse transcription-quantitative PCR
results showed the mRNA expression level of GRP-78 increases in
HSMCs treated with Ang II. **P<0.01,
***P<0.001 and ****P<0.0001. Ang II,
angiotensin II; HSCMs, human smooth muscle cells; DKK1, Dickkopf-1;
IL, interleukin; GRP, glucose regulated protein.

Figure 4

DEGs analysis after DKK1 silencing.
(A) The distribution of DEGs. (B) Volcano map of all DEGs. (C)
Volcano map of DEGs in NC-Ang II vs. NC. (D) Volcano map of DEGs in
siDKK1 vs. NC. (E) Volcano map of DEGs in siDKK1-Ang II vs. NC-Ang
II. (F) Clustering heat map of all DEGs. (G) Clustering heat map of
DEGs in NC-Ang II vs. NC. (H) Clustering heat map of DEGs in siDKK1
vs. NC. (I) Clustering heat map of DEGs in siDKK1-Ang II vs. NC-Ang
II. DEGs, differentially expressed genes; NC, treated with
scrambled siRNAs as negative control; siDKK1, treated with siRNAs
targeting human DKK1; Ang II, treated with Ang II (1 µM). NC,
negative control; siDKK1, small interfering RNA for DKK1; DKK1,
Dickkopf-1; Ang II, angiotensin II.

Figure 5

GO and KEGG functional enrichment
analysis. (A) Functional enrichment analysis of GO and KEGG of DEGs
in NC-Ang II vs. NC. (B) Functional enrichment analysis of GO and
KEGG of DEGs in siDKK1 vs. NC. (C) Functional enrichment analysis
of GO and KEGG of DEGs in siDKK1-Ang II vs. NC-Ang II. NC, treated
with scrambled siRNAs as negative control; siDKK1, treated with
siRNAs targeting human DKK1; Ang II, treated with Ang II (1 µM).
GO, gene ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes;
NC, negative control; siDKK1, small interfering RNA for DKK1; DKK1,
Dickkopf-1; Ang II, angiotensin II; BP, biological processes; CC,
cellular components; MF, molecular functions; P adj, adjusted
P.
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Copy and paste a formatted citation
Spandidos Publications style
Chang X, Yao B, Liu Z, Guo Z, Qiao P, Li X, Wang L and Li A: DKK1/SMDT1 participate in Ang II‑induced mitochondrial injury of human smooth muscle cells in an abdominal aortic aneurysm cell model. Exp Ther Med 31: 117, 2026.
APA
Chang, X., Yao, B., Liu, Z., Guo, Z., Qiao, P., Li, X. ... Li, A. (2026). DKK1/SMDT1 participate in Ang II‑induced mitochondrial injury of human smooth muscle cells in an abdominal aortic aneurysm cell model. Experimental and Therapeutic Medicine, 31, 117. https://doi.org/10.3892/etm.2026.13112
MLA
Chang, X., Yao, B., Liu, Z., Guo, Z., Qiao, P., Li, X., Wang, L., Li, A."DKK1/SMDT1 participate in Ang II‑induced mitochondrial injury of human smooth muscle cells in an abdominal aortic aneurysm cell model". Experimental and Therapeutic Medicine 31.4 (2026): 117.
Chicago
Chang, X., Yao, B., Liu, Z., Guo, Z., Qiao, P., Li, X., Wang, L., Li, A."DKK1/SMDT1 participate in Ang II‑induced mitochondrial injury of human smooth muscle cells in an abdominal aortic aneurysm cell model". Experimental and Therapeutic Medicine 31, no. 4 (2026): 117. https://doi.org/10.3892/etm.2026.13112
Copy and paste a formatted citation
x
Spandidos Publications style
Chang X, Yao B, Liu Z, Guo Z, Qiao P, Li X, Wang L and Li A: DKK1/SMDT1 participate in Ang II‑induced mitochondrial injury of human smooth muscle cells in an abdominal aortic aneurysm cell model. Exp Ther Med 31: 117, 2026.
APA
Chang, X., Yao, B., Liu, Z., Guo, Z., Qiao, P., Li, X. ... Li, A. (2026). DKK1/SMDT1 participate in Ang II‑induced mitochondrial injury of human smooth muscle cells in an abdominal aortic aneurysm cell model. Experimental and Therapeutic Medicine, 31, 117. https://doi.org/10.3892/etm.2026.13112
MLA
Chang, X., Yao, B., Liu, Z., Guo, Z., Qiao, P., Li, X., Wang, L., Li, A."DKK1/SMDT1 participate in Ang II‑induced mitochondrial injury of human smooth muscle cells in an abdominal aortic aneurysm cell model". Experimental and Therapeutic Medicine 31.4 (2026): 117.
Chicago
Chang, X., Yao, B., Liu, Z., Guo, Z., Qiao, P., Li, X., Wang, L., Li, A."DKK1/SMDT1 participate in Ang II‑induced mitochondrial injury of human smooth muscle cells in an abdominal aortic aneurysm cell model". Experimental and Therapeutic Medicine 31, no. 4 (2026): 117. https://doi.org/10.3892/etm.2026.13112
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