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Article Open Access

Atractylodin inhibits ferroptosis in sepsis‑induced acute gastrointestinal injury via SIRT3/PRDX3

  • Authors:
    • Yun-Xia Hu
    • Ming-Qi Chen
    • Jian-Lin Wang
    • Tao Wu
    • Hai-Dong Zhang
    • Tong-Tong Li
    • Hui Gao
    • Yu Bai
  • View Affiliations / Copyright

    Affiliations: Department of Intensive Care Medicine, Liyang Branch of Jiangsu Provincial Hospital of Chinese Medicine, Changzhou, Jiangsu 213300, P.R. China, Department of Intensive Care Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210004, P.R. China, Department of Intensive Care Medicine, Liyang Branch of Jiangsu Provincial Hospital of Chinese Medicine, Changzhou, Jiangsu 213300, P.R. China, Department of Intensive Care Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210004, P.R. China, Clinical Department of Integrated Traditional Chinese and Western Medicine, School of Integrative Medicine, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, P.R. China
    Copyright: © Hu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 330
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    Published online on: September 26, 2025
       https://doi.org/10.3892/mmr.2025.13695
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Abstract

Gastrointestinal injury (GI) is a significant concern in various medical contexts, particularly in patients undergoing antiplatelet therapy, experiencing trauma, or dealing with the effects of medications. The present study investigated the effect of atractylodin, a bioactive compound derived from Atractylodes lancea (Thunb.) DC., traditionally used in medicinal applications. A sepsis animal model was established through cecal ligation and perforation. Mice were treated with atractylodin, with or without silencing of NAD‑dependent protein deacetylase sirtuin‑3 (SIRT3), via transfection with adeno‑associated virus (AAV) vectors. Atractylodin markedly improved mitochondrial function in vivo, as evidenced by increased mitochondrial‑related proteins via western blot analysis (TOM20) and increased mitochondrial membrane potential, as observed via JC‑1 staining. In addition, atractylodin treatment inhibited apoptosis. Together, these changes regulated mitochondrial dysfunction. Moreover, atractylodin improved the prognosis of sepsis‑induced ferroptosis in the stomach and colon tissues. Atractylodin markedly activated SIRT3 while suppressing the expression of ac‑peroxiredoxin‑3 (PRDX3). Notably, the knockdown of SIRT3 diminishes the inhibitory effect of atractylodin on ferroptosis, when AAV‑short hairpinSIRT3 was injected into the stomach and colon tissues of C57 BL/6 mice. Further, atractylodin may have attenuated GI development by preventing mitochondrial dysfunction through the SIRT3/PRDX3 pathway. Hence, protection against mitochondrial dysfunction using atractylodin may be a promising therapeutic strategy against sepsis‑induced acute GI.
View Figures

Figure 1

Atractylodin mitigates sepsis-induced
GI. (A) H&E staining of mousee stomach (scale bar, 200 µm) and
colon tissue. (B) The immunohistochemical detection of tissue
damage in the stomach; scale bar, 100 µm. (C) Relative expression
of Occludin in the stomach. (D) Relative expression of ZO-1 in the
stomach. (E) The immunohistochemical detection of tissue damage in
the colon, scale bar 100 µm. (F) Relative expression of occludin in
the colon. (G) Relative expression of ZO-1 in the colon. The (H)
stomach tissue and (J) colon tissue cell apoptosis were analyzed
via TUNEL assay. Blue is nuclei stained with DAPI, while red is
TUNEL-positive cells stained with TUNEL; scale bar 50 µm. The
percentage of TUNEL-positive cells in (I) stomach tissues and (K)
colon tissues. TUNEL-positive cells (%) were calculated by the
number of positive cells divided by the total number of cells. Data
are combined from n=3 biological repeats. *P<0.05, **P<0.01
vs. the model group, ##P<0.01 vs. the Sham group. GI,
gastrointestinal injury; H&E, hematoxylin-eosin; TUNEL,
terminal dUTP nick end labeling; ZO-1, Zona occludens protein
1.

Figure 2

Effect of atractylodin on regulating
mitochondrial dysfunction. (A) Regulation of mitochondrial
oxidative damage markers: CAT, SOD 2, GSH/GSSG,
H2O2, MDA in the stomach, and (B) colon
tissue. (C) TEM images of mitochondrial morphology of each group.
Western blot analysis of mitochondrial membrane TOM20 markers in
(D) stomach and (E) colon tissue. Scale bar 500 nm. Relative
quantitation of the intensity of TOM20 in the (F) stomach and (G)
colon tissue. Values are means ± SD of n=6 (A, B, F and G) or n=3
(D and E) experiments. *P<0.05, **P<0.01 vs. the model group,
##P<0.01 vs. the Sham group. CAT, catalase; SOD 2,
superoxide dismutase; GSH/GSSG, glutathione/glutathione (oxidized);
MDA, malondialdehyde TEM, transmission electron microscopy; TOM20,
mitochondrial import receptor subunit TOM20 homolog; MDA,
malondialdehyde.

Figure 3

Results of stomach and colon tissue
mitochondrial membrane potential and ROS. Flow cytometric diagram
of cell mitochondrial membrane potential in each group in (A)
stomach and (C) colon tissue. Green fluorescence represents the
monomeric form of JC-1 (JC-1 monomers), indicating mitochondrial
transmembrane potential dissipation. Analysis of cell mitochondrial
membrane potential in each group in (B) stomach and (D) colon
tissue. DHE fluorescence (red) images in the (E) stomach and (G)
colon tissue; scale bar 50 µm. Statistical analysis results of
mitochondrial ROS in (F) stomach and (H) colon tissue. The activity
of mitochondrial respiratory chain complex I, II, III, IV, V in (I)
stomach and (J) colon tissue. Values are means ± SD of n=3 (A-H) or
n=6 (I and J) experiments. *P<0.05, **P<0.01 vs. the model
group, ##P<0.01 vs. the Sham group. ROS, reactive
oxygen species; DHE, dihydroethidium.

Figure 4

Atractylodin treatment improves the
prognosis of sepsis-induced ferroptosis in the stomach and colon
tissues. (A) Determination of the expression levels of
Fe2+ in stomach tissues. (B) Protein expression levels
of GPX4, SLC7A11, TFR1 and GAPDH in stomach tissue in each group.
Quantification of relative protein expression of (C) GPX4, (D)
SLC7A11 and (E) TFR1in the stomach tissue. (F) Determination of the
expression levels of Fe2+ in colon tissues. (G) Protein
expression levels of GPX4, SLC7A11, TFR1 and GAPDH in colon tissue
in each group. (H-I) Quantifying relative protein expression of (H)
GPX4, (I) SLC7A11 and (J) TFR1 in the colon tissue. Values are
means ± SD of n=6 (A and F) or n=3 (B-E and G-J) experiments.
*P<0.05, **P<0.01 vs. the model group, ##P<0.01
vs. the Sham group. GPX4, Phospholipid hydroperoxide glutathione
peroxidase; SLC7A11, Solute carrier family 7 member 11; TFR1,
transferrin receptor protein 1; GAPDH, glyceraldehyde-3-phosphate
dehydrogenase.

Figure 5

Atractylodin suppresses the
expression of Ac-PRDX3 while inducing the expression of SIRT3 in
stomach and colon tissues. (A) The western blot assay of Ac-PRDX3
and PRDX3 in stomach tissue and (E) colon tissue of each group.
Quantifying relative protein expression of Ac-PRDX3/PRDX3 in the
(B) stomach and (F) colon tissue. The western blot assay of SIRT3
in (C) stomach and (G) colon tissue of each group. Quantification
of relative protein expression of SIRT3/GAPDH in the (D) stomach
and (H) colon tissue. Confocal immunofluorescence of the mice
cochlear tissues in different groups. Confocal immunofluorescence
of SIRT3 (green) in (I) stomach tissue and (K) colon tissue; scale
bar, 50 µm. the cell nuclear parts were labeled with blue.
Quantification of relative expression of SIRT3 in the (J) stomach
and (L) colon tissue. Co-IP of PRDX3 with SIRT3 in (M) stomach and
(N) colon tissue. (O-R) Relative quantitative evaluation of the
western-blot analysis for SIRT3/PRDX3, PRDX3/GAPDH expression was
obtained using ImageJ software. Values are means ± SD of n=3 (A-R)
experiments. *P<0.05, **P<0.01 vs. the model group,
##P<0.01 vs. the Sham group. PRDX3, peroxiredoxin-3;
SIRT3, NAD-dependent protein deacetylase sirtuin-3, mitochondrial;
Co-IP, coimmunoprecipitation.

Figure 6

Regulatory mechanism of SIRT3 and
PRDX3 in sepsis-induced ferroptosis-mediated mitochondrial
oxidative damage. (A) Western blot analysis of SIRT3 protein level
in the stomach and colon tissue. Mice were injected with sh-SIRT3
or control shRNA (shRNA-NC). (B) Western blot analysis of GPX4
protein level in stomach and (C) Western blot analysis of GPX4
protein level in colon tissue. (D) Relative SIRT3/GAPDH expression
obtained using ImageJ software. Relative quantitative evaluation of
the GPX4/GAPDH expression in (E) stomach and (F) colon tissue of
each group. (G) Western blot analysis of TOM20, occludin, and ZO-1
protein level in stomach tissue of each group. Relative
quantitative evaluation of (H) TOM20, (I) occludin and (J) ZO-1
protein expression level in stomach tissue in each group. (K)
Western blot analysis of TOM20, occludin, and ZO-1 protein level in
stomach and colon tissue of each group. Relative quantitative
evaluation of (L) TOM20, (M) occludin and (N) ZO-1 protein
expression level in colon tissue. Activity of mitochondrial
respiratory chain complex (O) I, mitochondrial respiratory chain
complex II (P), mitochondrial respiratory chain complex III (Q),
mitochondrial respiratory chain complex IV (R), mitochondrial
respiratory chain complex V (S) in the stomach tissue. (T-X) The
activity of mitochondrial respiratory chain complex I (T),
mitochondrial respiratory chain complex II (U), mitochondrial
respiratory chain complex III (V), mitochondrial respiratory chain
complex IV (W), mitochondrial respiratory chain complex V (X) in
the colon tissue. Values are means ± SD of n=3 (A-N) or n=6 (O-X)
experiments. **P<0.01. SIRT3, NAD-dependent protein deacetylase
sirtuin-3, mitochondrial; PRDX3, peroxiredoxin-3; sh, short
hairpin; GPX4, Phospholipid hydroperoxide glutathione peroxidase;
TOM20, mitochondrial import receptor subunit TOM20 homolog; ZO-1,
zona occludens protein 1.
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Copy and paste a formatted citation
Spandidos Publications style
Hu Y, Chen M, Wang J, Wu T, Zhang H, Li T, Gao H and Bai Y: Atractylodin inhibits ferroptosis in sepsis‑induced acute gastrointestinal injury via SIRT3/PRDX3. Mol Med Rep 32: 330, 2025.
APA
Hu, Y., Chen, M., Wang, J., Wu, T., Zhang, H., Li, T. ... Bai, Y. (2025). Atractylodin inhibits ferroptosis in sepsis‑induced acute gastrointestinal injury via SIRT3/PRDX3. Molecular Medicine Reports, 32, 330. https://doi.org/10.3892/mmr.2025.13695
MLA
Hu, Y., Chen, M., Wang, J., Wu, T., Zhang, H., Li, T., Gao, H., Bai, Y."Atractylodin inhibits ferroptosis in sepsis‑induced acute gastrointestinal injury via SIRT3/PRDX3". Molecular Medicine Reports 32.6 (2025): 330.
Chicago
Hu, Y., Chen, M., Wang, J., Wu, T., Zhang, H., Li, T., Gao, H., Bai, Y."Atractylodin inhibits ferroptosis in sepsis‑induced acute gastrointestinal injury via SIRT3/PRDX3". Molecular Medicine Reports 32, no. 6 (2025): 330. https://doi.org/10.3892/mmr.2025.13695
Copy and paste a formatted citation
x
Spandidos Publications style
Hu Y, Chen M, Wang J, Wu T, Zhang H, Li T, Gao H and Bai Y: Atractylodin inhibits ferroptosis in sepsis‑induced acute gastrointestinal injury via SIRT3/PRDX3. Mol Med Rep 32: 330, 2025.
APA
Hu, Y., Chen, M., Wang, J., Wu, T., Zhang, H., Li, T. ... Bai, Y. (2025). Atractylodin inhibits ferroptosis in sepsis‑induced acute gastrointestinal injury via SIRT3/PRDX3. Molecular Medicine Reports, 32, 330. https://doi.org/10.3892/mmr.2025.13695
MLA
Hu, Y., Chen, M., Wang, J., Wu, T., Zhang, H., Li, T., Gao, H., Bai, Y."Atractylodin inhibits ferroptosis in sepsis‑induced acute gastrointestinal injury via SIRT3/PRDX3". Molecular Medicine Reports 32.6 (2025): 330.
Chicago
Hu, Y., Chen, M., Wang, J., Wu, T., Zhang, H., Li, T., Gao, H., Bai, Y."Atractylodin inhibits ferroptosis in sepsis‑induced acute gastrointestinal injury via SIRT3/PRDX3". Molecular Medicine Reports 32, no. 6 (2025): 330. https://doi.org/10.3892/mmr.2025.13695
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