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Article Open Access

Histone deacetylase 3‑mediated histone deacetylation combined with activating transcription factor 3 promotes renal fibrosis by inhibiting Klotho

  • Authors:
    • Zhiying Yang
    • Xueling Wang
    • Wenhong Li
    • Hao Chen
    • Gang Liu
  • View Affiliations / Copyright

    Affiliations: Department of Nephrology, Yan'an Hospital Affiliated to Kunming Medical University, Kunming, Yunnan 650051, P.R. China, Department of Nephrology, The Second Qilu Hospital of Shandong University, Jinan, Shandong 250033, P.R. China
    Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 274
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    Published online on: August 6, 2026
       https://doi.org/10.3892/mmr.2026.13985
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Abstract

Renal fibrosis is a representative pathological trait of renal aging and chronic kidney disease. However, the regulatory mechanisms of histone deacetylation in renal fibrosis remain unclear. A mouse renal fibrosis model was constructed using the unilateral ureteric obstruction method, and HK2 cells treated with TGF‑β were used to create a renal fibrosis cell model. Low expression of Klotho in renal fibrosis is associated with histone deacetylase 3 (HDAC3)‑mediated histone deacetylation. After TGF‑β treatment, H3K9ac and Klotho binding was markedly decreased, while H3K9ac was enriched in the Klotho group after the addition of the HDAC3 inhibitor. Further experiments demonstrated that HDAC3 binding activates transcription factor 3, inhibits Klotho transcription and promotes cellular renal fibrosis. The absence of Klotho may activate the Wnt/β‑catenin and NF‑κB pathways to promote oxidative stress and inflammation, thereby exacerbating the fibrotic process. HDAC3 binds to ATF3 to transcriptionally repress Klotho, leading to activation of the Wnt/β‑catenin and NF‑κB pathways and exacerbation of renal fibrosis.
View Figures

Figure 1

Expression of HDAC3 and Klotho in
kidney tissues from patients with CKD and their correlation. (A)
Klotho mRNA expression levels in clinical samples were assessed by
RT-qPCR; (B) HDAC3 mRNA levels in clinical samples were checked by
RT-qPCR; (C) Correlation analysis between Klotho and HDAC3 at the
clinical level. ***P<0.001. RT-qPCR, reverse
transcription-quantitative PCR.

Figure 2

Low expression of Klotho in renal
fibrosis may be associated with HDAC3-mediated histone
deacetylation. (A) Expression levels of Klotho in renal tissues of
mice were evaluated by immunohistochemical analysis. (B) Klotho and
(C) HDAC3 protein levels in renal tissues of mice were detected by
western blotting. (D) The levels of acetylated histone H3 in renal
tissues of mice was measured using a kit. (E) Klotho and (F) HDAC3
protein expression levels in HK2 cells were detected by western
blotting. (G) The expression levels of acetylated histone H3 in HK2
cells was measured using a kit. Data were analyzed using one-way
ANOVA followed by Tukey's post hoc test. *P<0.05, **P<0.01,
***P<0.001 vs. the control/sham/NC group. UUO, unilateral
ureteric obstruction; NC, negative control.

Figure 3

HDAC3 inhibits Klotho transcription
and promotes HK2 cell fibrosis through histone deacetylation. (A)
H3K9ac levels in the Klotho promoter region were detected by
ChIP-qPCR. (B) Klotho mRNA expression levels in HK2 cells were
detected by RT-qPCR. (C) Klotho protein levels in HK2 cells were
detected by western blotting. (D) Validation of Klotho knockdown
efficiency by siRNA in HK2 cells. (E) Collagen I, Collagen IV,
Fibronectin and α-SMA protein expression levels in HK2 cells were
detected by western blotting. Data were analyzed using one-way
ANOVA followed by Tukey's post hoc test. **P<0.01, ***P<0.001
vs. NC group; #P<0.05, ##P<0.01,
###P<0.001 vs. the TGF-β group;
△P<0.05, △△P<0.01 vs. the TGF-β+RGFP966
group. ChIP, chromatin immunoprecipitation; RT-qPCR, reverse
transcription-quantitative PCR; siRNA, small interfering RNA;
α-SMA, α-smooth muscle actin; NC, negative control.

Figure 4

HDAC3 binding to ATF3
transcriptionally downregulates Klotho. (A) ATF3 protein expression
in renal tissue of mice was detected by western blotting. (B) ATF3
protein expression in HK2 cells was confirmed by western blotting.
The binding of (C) HDAC3 and (D) ATF3 was corroborated by
coimmunoprecipitation in HK2 cells and renal tissue of mice. (E)
The binding of ATF3 to the Klotho promoter region was validated by
ChIP-qPCR. (F) Validation of ATF3 knockdown efficiency by siRNA in
HK2 cells. (G) ATF3 mRNA expression in HK2 cells was distinguished
by RT-qPCR. (H) Klotho protein expression in HK2 cells was detected
by western blotting. (I) Klotho protein expression in renal tissue
of mice was identified by western blotting. (J) Collagen I and IV,
Fibronectin and α-SMA protein expression in renal tissue of mice
were confirmed by western blotting. (K) Mouse pathological sections
were observed by Masson's Trichrome staining. Data were analyzed
using one-way ANOVA followed by Tukey's post hoc test. *P<0.05,
***P<0.001 vs. the NC/sham group; ##P<0.01,
###P<0.001 vs. the TGF-β group. ATF3, activates
transcription factor 3; UUO, unilateral ureteric obstruction; ChIP,
chromatin immunoprecipitation; RT-qPCR, reverse
transcription-quantitative PCR; siRNA, small interfering RNA;
α-SMA, α-smooth muscle actin; NC, negative control.

Figure 5

ATF3 mediates the inhibitory effect
of Klotho on Wnt/β-catenin signaling pathway. (A) Wnt1, β-catenin,
c-MYC and Cyclin D1 protein expression in HK2 cells was detected by
western blotting. (B) Collagen I, Collagen IV, Fibronectin and
α-SMA protein expression in HK2 cells was identified by western
blotting. (C) HDAC3 protein expression in HK2 cells was detected by
western blotting. (D) Wnt1, β-catenin, c-MYC and Cyclin D1 proteins
in renal tissue of mice were detected by western blotting. (E)
Collagen I, Collagen IV, Fibronectin and α-SMA protein expression
in renal tissue of mice was detected by western blotting. (F) HDAC3
protein expression in renal tissue of mice was identified by
western blotting. (G) Mouse pathological section was observed by
Masson's Trichrome staining. Data were analyzed using one-way ANOVA
followed by Tukey's post hoc test. **P<0.01, ***P<0.001 vs.
the NC/sham group; ###P<0.001 vs. the TGF-β/UUO
group; △P<0.05, △△P<0.01 vs. the TGF-β
+ oe-Klotho/UUO + oe-Klotho group. NC, negative control; c-MYC,
cellular myelocytomatosis viral oncogene homolog; ATF3, activates
transcription factor 3; UUO, unilateral ureteric obstruction;
α-SMA, α-smooth muscle actin; HDAC3, histone deacetylase 3; oe,
overexpression.

Figure 6

ATF3 mediates the inhibitory effect
of Klotho on NF-κB signaling pathway. (A) NF-κB p65 protein
expression in HK2 cells was detected by western blotting. (B)
Nuclear translocation of NF-κB p65 in HK2 cells was observed by
immunofluorescence. (C) SOD and (D) MDA levels in HK2 cells were
tested by kits. (E) ROS levels in HK2 cells were tested by flow
cytometry. (F) NF-κB p65 protein expression in renal tissue of mice
was detected by western blotting. (G) TNF-α, IL-1β and IL-6 in
mouse serum were measured by ELISA kits. Data were analyzed using
one-way ANOVA followed by Tukey's post hoc test. ***P<0.001 vs.
the NC/sham group; ###P<0.001 vs. the TGF-β/UUO
group; △P<0.05, △△P<0.01 vs. the TGF-β
+ oe-Klotho/UUO + oe-Klotho group. ATF3, activates transcription
factor 3; NC, negative control; UUO, unilateral ureteric
obstruction; oe, overexpression.

Figure 7

Mechanism diagram. Under the
stimulation of TGF-β on renal tubular epithelial cells, the H3K9ac
modification mediated by the binding of HDAC3 and ATF3 inhibits the
transcription of Klotho. The downregulation of Klotho may further
activate the Wnt/β-catenin and NF-κB pathways, thereby exacerbating
renal fibrosis. HDAC3, histone deacetylase 3; ATF3, activates
transcription factor 3.
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Copy and paste a formatted citation
Spandidos Publications style
Yang Z, Wang X, Li W, Chen H and Liu G: Histone deacetylase 3‑mediated histone deacetylation combined with activating transcription factor 3 promotes renal fibrosis by inhibiting Klotho. Mol Med Rep 34: 274, 2026.
APA
Yang, Z., Wang, X., Li, W., Chen, H., & Liu, G. (2026). Histone deacetylase 3‑mediated histone deacetylation combined with activating transcription factor 3 promotes renal fibrosis by inhibiting Klotho. Molecular Medicine Reports, 34, 274. https://doi.org/10.3892/mmr.2026.13985
MLA
Yang, Z., Wang, X., Li, W., Chen, H., Liu, G."Histone deacetylase 3‑mediated histone deacetylation combined with activating transcription factor 3 promotes renal fibrosis by inhibiting Klotho". Molecular Medicine Reports 34.4 (2026): 274.
Chicago
Yang, Z., Wang, X., Li, W., Chen, H., Liu, G."Histone deacetylase 3‑mediated histone deacetylation combined with activating transcription factor 3 promotes renal fibrosis by inhibiting Klotho". Molecular Medicine Reports 34, no. 4 (2026): 274. https://doi.org/10.3892/mmr.2026.13985
Copy and paste a formatted citation
x
Spandidos Publications style
Yang Z, Wang X, Li W, Chen H and Liu G: Histone deacetylase 3‑mediated histone deacetylation combined with activating transcription factor 3 promotes renal fibrosis by inhibiting Klotho. Mol Med Rep 34: 274, 2026.
APA
Yang, Z., Wang, X., Li, W., Chen, H., & Liu, G. (2026). Histone deacetylase 3‑mediated histone deacetylation combined with activating transcription factor 3 promotes renal fibrosis by inhibiting Klotho. Molecular Medicine Reports, 34, 274. https://doi.org/10.3892/mmr.2026.13985
MLA
Yang, Z., Wang, X., Li, W., Chen, H., Liu, G."Histone deacetylase 3‑mediated histone deacetylation combined with activating transcription factor 3 promotes renal fibrosis by inhibiting Klotho". Molecular Medicine Reports 34.4 (2026): 274.
Chicago
Yang, Z., Wang, X., Li, W., Chen, H., Liu, G."Histone deacetylase 3‑mediated histone deacetylation combined with activating transcription factor 3 promotes renal fibrosis by inhibiting Klotho". Molecular Medicine Reports 34, no. 4 (2026): 274. https://doi.org/10.3892/mmr.2026.13985
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