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HSPD1 regulates angiotensin II‑induced atrial fibrillation via the PPAR signaling pathway

  • Authors:
    • Yimeng Zhou
    • Lianzhi Zhang
    • Shunping Zhou
    • Wenjia Zhang
    • Qunlin Gong
    • Nan Xu
    • Jiahong Wang
    • Zhong Zhang
    • Nannan Chen
  • View Affiliations / Copyright

    Affiliations: Department of Cardiology, School of Medicine, Yangpu Hospital, Tongji University, Shanghai 200090, P.R. China
    Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 331
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    Published online on: September 26, 2025
       https://doi.org/10.3892/mmr.2025.13696
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Abstract

Atrial fibrillation (AF) is a multifactorial condition, and understanding its molecular mechanisms is key for developing targeted therapies. The present study aimed to investigate the role of heat shock protein D1 (HSPD1) in the pathogenesis of AF, particularly in angiotensin II (Ang II)‑induced cardiac fibroblasts (CFs). Differentially expressed genes (DEGs) were screened from the GSE31821 dataset for enrichment analysis, protein‑protein interaction (PPI) network construction and the hub gene HSPD1 was identified. In vitro experiments, including Cell Counting Kit‑8 and Transwell migration assay, Reverse transcription‑quantitative (RT‑q)PCR, Western blotting and ELISA, were conducted to evaluated the effects of Ang II on CF viability, migration, fibrosis‑associated markers (collagen I, collagen III and α‑smooth muscle actin) and HSPD1 expression. The effects of HSPD1 knockdown on cell viability, inflammatory cytokines (TNF‑α, IL‑8 and IL‑1β), AF‑associated proteins [atrial natriuretic peptide (ANP), β‑major histocompatibility complex (MHC) and MMP‑2] and peroxisome proliferator‑activated receptor (PPAR) signaling pathways were investigated. A total of 582 DEGs were identified, with significant involvement of pathways such as ‘MAPK signaling pathway’ and ‘Wnt signaling pathway’. A total of five genes (HSP90AA1, HSP90AB1, HSPA4, HSPA8 and HSPD1) were highly expressed in AF samples. In vitro, Ang II‑induced fibrotic changes in CFs included increased viability, migration and upregulated fibrosis markers and HSPD1 expression. HSPD1 knockdown decreased Ang II‑induced secretion of inflammatory cytokines (TNF‑α, IL‑8 and IL‑1β) and expression of AF‑associated proteins (ANP, β‑MHC and MMP‑2). Downregulation of PPAR signaling proteins (PPARα, PPARγ, carnitine palmitoyltransferase I and sirtuin3) were observed, while thiazolidinedione treatment partially prevented Ang II‑induced changes. HSPD1 serves a key role in Ang II‑induced CF function, promoting fibrosis, inflammation and AF‑associated protein expression. Targeting HSPD1 in combination with PPAR pathway modulation presents a promising therapeutic strategy for AF treatment.
View Figures

Figure 1

Screening and enrichment analysis of
GSE31821 DEGs. (A) Volcano plot of filtered DEGs from the GSE31821
dataset. Enrichment analysis of DEGs of the GSE31812 dataset,
including (B) biological process, (C) cellular component, (D)
molecular function and (E) KEGG. DEG, differentially expressed
gene; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Figure 2

Expression analysis of candidate
genes in the PPI network. PPI network of DEGs of the GSE31812
dataset, including (A) MCC, (B) MNC and (C) degree. (D) Venn
diagram of the intersection of MCC, MNC and degree. Expression
analysis of five overlapping genes in (E) GSE31821 dataset and (F)
GSE14975 dataset. *P<0.05. PPI, protein-protein interaction;
DEG, differentially expressed gene; MCC, maximum correlation
criterion; MNC, maximum neighborhood component; AF, atrial
fibrillation; HSP, heat shock protein.

Figure 3

Effects of Ang II induction on
fibrosis in CFs. (A) Viability of CFs treated with different
concentrations of Ang II for 24 h, measured using the Cell Counting
Kit-8 assay. *P<0.05 vs. the Control group. (B) Cell viability
of CFs treated with 1 µM Ang II for 0, 6, 12, 24, and 48 h.
*P<0.05 vs. the 1 µM Ang II for 0 h group. (C) Transwell assay
of CF migration after induction with 1 µM Ang II for 48 h. Scale
bar, 50 µm. *P<0.05 vs. the Control group. (D) mRNA and (E)
protein expression of HSPD1 in CF cells induced by 1 µM Ang
II for 48 h. *P<0.05 vs. the Control group. (F) mRNA expression
of collagen I, collagen III and α-SMA in CFs following induction
with 1 µM Ang II for 48 h. *P<0.05 vs. the Control group. (G)
Representative western blotting and (H) protein expression of
collagen I, collagen III and α-SMA in CFs following induction with
1 µM Ang II for 48 h. *P<0.05 vs. Control group Ang II,
angiotensin II; CF, cardiac fibroblast; HSPD1, heat shock protein
D1.

Figure 4

Downregulation of HSPD1 improves Ang
II-induced AF. (A) mRNA and (B) protein expression of HSPD1 in
HSPD1-knockdown CFs. *P<0.05 vs. the si-NC group. (C)
mRNA and (D) protein expression of HSPD1 in CFs. *P<0.05 vs. the
Ang II+ si-NC group. (E) Cell viability of CFs was assessed by Cell
Counting Kit-8 assay. ELISA was used to detect the secretion of
inflammatory factors (F) TNF-α, (G) IL-8 and (H) IL-1β in CFs. (I)
Representative western blotting and protein expression of the
AF-associated markers (J) ANP, (K) β-MHC and (L) MMP-2 in CFs.
*P<0.05 vs. the Control group; #P<0.05 vs. the Ang
II+ si-NC group. AF, atrial fibrillation; Ang II, angiotensin II;
CF, cardiac fibroblast; si, small interfering; NC, negative
control; HSPD1, heat shock protein D1; ANP, atrial natriuretic
peptide; MHC, major histocompatibility complex.

Figure 5

Regulation of Ang II-induced AF by
HSPD1 via the PPAR signaling pathway. (A) Representative western
blotting and protein expression of PPAR signaling
pathway-associated proteins (B) PPARα, (C) PPARγ, (D) CPT-1 and (E)
SIRT3 in CFs. (F) Cell Counting Kit-8 assay of CF viability. ELISA
detection of the expression of the inflammatory factors (G) TNF-α,
(H) IL-8 and (I) IL-1β in CFs. (J) Representative western blotting
and protein expression of the AF markers (K) ANP, (L) β-MHC and (M)
MMP-2 in CFs. *P<0.05 vs. Ang II+ si-NC; #P<0.05
vs. the Ang II+si-HSPD1-2 group. AF, atrial fibrillation; TZD,
thiazolidinedione; CF, cardiac fibroblast; si, small interfering;
NC, negative control; PPAR, peroxisome proliferator-activated
receptor; CPT-1, carnitine palmitoyltransferase I; SIRT, sirtuin;
ANP, atrial natriuretic peptide; MHC, major histocompatibility
complex.
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Copy and paste a formatted citation
Spandidos Publications style
Zhou Y, Zhang L, Zhou S, Zhang W, Gong Q, Xu N, Wang J, Zhang Z and Chen N: HSPD1 regulates angiotensin II‑induced atrial fibrillation via the PPAR signaling pathway. Mol Med Rep 32: 331, 2025.
APA
Zhou, Y., Zhang, L., Zhou, S., Zhang, W., Gong, Q., Xu, N. ... Chen, N. (2025). HSPD1 regulates angiotensin II‑induced atrial fibrillation via the PPAR signaling pathway. Molecular Medicine Reports, 32, 331. https://doi.org/10.3892/mmr.2025.13696
MLA
Zhou, Y., Zhang, L., Zhou, S., Zhang, W., Gong, Q., Xu, N., Wang, J., Zhang, Z., Chen, N."HSPD1 regulates angiotensin II‑induced atrial fibrillation via the PPAR signaling pathway". Molecular Medicine Reports 32.6 (2025): 331.
Chicago
Zhou, Y., Zhang, L., Zhou, S., Zhang, W., Gong, Q., Xu, N., Wang, J., Zhang, Z., Chen, N."HSPD1 regulates angiotensin II‑induced atrial fibrillation via the PPAR signaling pathway". Molecular Medicine Reports 32, no. 6 (2025): 331. https://doi.org/10.3892/mmr.2025.13696
Copy and paste a formatted citation
x
Spandidos Publications style
Zhou Y, Zhang L, Zhou S, Zhang W, Gong Q, Xu N, Wang J, Zhang Z and Chen N: HSPD1 regulates angiotensin II‑induced atrial fibrillation via the PPAR signaling pathway. Mol Med Rep 32: 331, 2025.
APA
Zhou, Y., Zhang, L., Zhou, S., Zhang, W., Gong, Q., Xu, N. ... Chen, N. (2025). HSPD1 regulates angiotensin II‑induced atrial fibrillation via the PPAR signaling pathway. Molecular Medicine Reports, 32, 331. https://doi.org/10.3892/mmr.2025.13696
MLA
Zhou, Y., Zhang, L., Zhou, S., Zhang, W., Gong, Q., Xu, N., Wang, J., Zhang, Z., Chen, N."HSPD1 regulates angiotensin II‑induced atrial fibrillation via the PPAR signaling pathway". Molecular Medicine Reports 32.6 (2025): 331.
Chicago
Zhou, Y., Zhang, L., Zhou, S., Zhang, W., Gong, Q., Xu, N., Wang, J., Zhang, Z., Chen, N."HSPD1 regulates angiotensin II‑induced atrial fibrillation via the PPAR signaling pathway". Molecular Medicine Reports 32, no. 6 (2025): 331. https://doi.org/10.3892/mmr.2025.13696
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