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Article Open Access

Yuanhuacine modulates lipopolysaccharide‑induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury

  • Authors:
    • Ui Jeong Park
    • Jae Wha Kim
  • View Affiliations / Copyright

    Affiliations: Immune Therapy Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Republic of Korea
    Copyright: © Park et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 168
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    Published online on: July 3, 2025
       https://doi.org/10.3892/etm.2025.12918
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Abstract

Inflammation is an immune response that activates immune cells to protect the host from infection or tissue damage; however, excessive inflammation can lead to sepsis and acute kidney injury (AKI). Yuanhuacine (YC), a physiologically active compound derived from Daphne genkwa flowers, has demonstrated its therapeutic potential in various diseases, including inflammatory diseases and cancer. However, the underlying molecular mechanisms by which YC regulates inflammatory cytokines and exerts efficacy against AKI remain to be elucidated. The present study aimed to investigate the role of YC in regulating cytokines in human macrophages and to evaluate its protective effect in a mouse model of AKI. Lipopolysaccharide (LPS) was used to stimulate THP‑1 macrophages in vitro, and LPS was administered intraperitoneally to establish an in vivo AKI model. LPS treatment significantly increased interleukin 6 (IL‑6) expression in both macrophages and in mice with AKI. However, YC treatment effectively reduced IL‑6 production by inhibiting the activation of Janus kinase 1 (JAK1) and signal transducer and activator of transcription 3 (STAT3) in macrophages, and YC was confirmed to inhibit LPS‑induced tubular damage in the mouse model of AKI. In conclusion, YC may serve as a potential therapeutic agent in the prevention of AKI and other IL‑6‑related inflammatory diseases by promoting JAK1/STAT3 dephosphorylation to facilitate inflammation resolution.
View Figures

Figure 1

Structure of YC and its effect on
inflammatory cytokines in differentiated THP-1 cells. (A) Chemical
structure of YC. (B) Differentiated THP-1 cells were treated with
various concentrations of YC, with or without LPS for 24 h. (C)
Time-course analysis of inflammatory cytokine mRNA levels (0-8 h).
(D) mRNA levels of LPS-induced inflammatory cytokines after 4 h
treatment with various doses of YC. (E) Time-course analysis of
IL-6 protein levels (0-24 h). (F) IL-6 protein levels after
treatment with various concentrations of YC. Cell viability was
assessed using the Cell Counting Kit-8 assay. The mRNA and protein
levels of cytokines were analyzed using reverse transcription
polymerase chain reaction and enzyme-linked immunosorbent assay.
LPS (5 ng/ml) was used in all experiments. Data are presented as
the mean ± standard deviation from three independent experiments.
Statistical significance was determined using one-way and two-way
analysis of variance. ###P<0.001 vs. control;
$$$P<0.001, $P<0.05 vs. LPS; ns, not
significant; YC, yuanhuacine; LPS, lipopolysaccharide; IL,
interleukin.

Figure 2

YC inhibits LPS-induced
phosphorylation of JAK1 and STAT3 in THP-1 cells. (A)
Differentiated THP-1 cells were treated with 5 ng/ml LPS and 200 nM
YC for up to 12 h. The activation of kinases as well as
transcription factors was evaluated by western blotting analysis
following LPS stimulation. (B) Quantification of phosphorylated
JAK1 and STAT3 proteins after treatment with increasing
concentrations of YC. White lines indicate separate groups.
Phosphorylation levels were normalized to total JAK1 and STAT3
expression and quantified using ImageJ software. Data are presented
as the mean ± standard deviation from three independent
experiments. Statistical significance was determined using one-way
analysis of variance. ###P<0.001,
##P<0.01 vs. control; $$P<0.01 vs. LPS;
ns, not significant; YC, yuanhuacine; LPS, lipopolysaccharide;
JAK1, Janus kinase 1; STAT3, signal transducer and activator of
transcription 3; p-, phosphorylated.

Figure 3

YC attenuates LPS-induced
cytoplasmic-to-nuclear translocation of phosphorylated STAT3 in
THP-1 cells. THP-1 cells were treated with 5 ng/ml LPS for 2 h in
the absence or presence of YC at three concentrations: 8, 40, and
200 nM. The subcellular localization of STAT3 was assessed by
multiple methods. (A) Western blot analysis of cytoplasmic and
nuclear fractions was performed to determine the localization of
STAT3 protein. (B) Confocal microscopy image showing the
intracellular distribution of p-STAT3. (C) Confocal microscopy
image showing the intracellular distribution of total STAT3.
β-Actin was used as a cytoplasmic marker, and Lamin B served as a
nuclear marker. Nuclei were stained with DAPI (blue), and STAT3
(total and p-STAT3) was visualized in green. Scale bars: 10 µm. YC,
yuanhuacine; LPS, lipopolysaccharide; STAT3, signal transducer and
activator of transcription 3; p-, phosphorylated.

Figure 4

Effect of YC on IL-6 expression
compared with Janus kinase 1 (filgotinib) and signal transducer and
activator of transcription 3 (S3I-201) inhibitors. THP-1 cells were
treated with 5 ng/ml LPS for 24 h in the presence of varying
concentrations of (A) filgotinib, (B) S3I-201 and (C) YC. IL-6
levels were measured in the supernatants of the treated cells using
enzyme-linked immunosorbent assay. Data are presented as the mean ±
standard deviation from three independent experiments. Statistical
differences were analyzed using one-way analysis of variance.
###P<0.001, ##P<0.01 vs. control;
$$$P<0.001, $$P<0.01 vs. LPS; ns, not
significant; YC, yuanhuacine; LPS, lipopolysaccharide; IL,
interleukin.

Figure 5

Protective effects of YC in an
LPS-induced AKI mouse model. Renal dysfunction was evaluated by
measuring (A) BUN and (B) sCr levels, and (C) tubular injury was
assessed by scoring histological damage (n=5). (D) A representative
image of H&E staining and immunohistochemistry images in kidney
tissue. The arrows indicate tubular degeneration. In kidney tissues
of mice with AKI (n=5), the expression levels of (E) KIM-1, (F)
NGAL and (G) IL-6, markers of renal injury, were analyzed by
quantitative polymerase chain reaction. (H) Western blot analysis
of JAK1 and STAT3 activation in kidney tissues of AKI mice (n=4)
following YC treatment. The image density was analyzed using
ImageJ. Scale bars: 500 µm (40x), 100 µm (100x, 200x), and 50 µm
(400x). Data are presented as the mean ± standard deviation.
Statistical significance was determined using one-way analysis of
variance. ###P<0.001, ##P<0.01,
#P<0.05 vs. control; $$$P<0.001,
$$P<0.01, $P<0.05 vs. LPS; ns, not
significant; AKI, acute kidney injury; H&E, hematoxylin and
eosin; KIM-1, kidney injury molecule 1; NGAL, neutrophil
gelatinase-associated lipocalin; BUN, blood urea nitrogen; sCr,
serum creatinine; YC, yuanhuacine; LPS, lipopolysaccharide; IL-6,
interleukin-6; JAK1, Janus kinase 1; STAT3, signal transducer and
activator of transcription 3.
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Copy and paste a formatted citation
Spandidos Publications style
Park U and Kim J: Yuanhuacine modulates lipopolysaccharide‑induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury. Exp Ther Med 30: 168, 2025.
APA
Park, U., & Kim, J. (2025). Yuanhuacine modulates lipopolysaccharide‑induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury. Experimental and Therapeutic Medicine, 30, 168. https://doi.org/10.3892/etm.2025.12918
MLA
Park, U., Kim, J."Yuanhuacine modulates lipopolysaccharide‑induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury". Experimental and Therapeutic Medicine 30.3 (2025): 168.
Chicago
Park, U., Kim, J."Yuanhuacine modulates lipopolysaccharide‑induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury". Experimental and Therapeutic Medicine 30, no. 3 (2025): 168. https://doi.org/10.3892/etm.2025.12918
Copy and paste a formatted citation
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Spandidos Publications style
Park U and Kim J: Yuanhuacine modulates lipopolysaccharide‑induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury. Exp Ther Med 30: 168, 2025.
APA
Park, U., & Kim, J. (2025). Yuanhuacine modulates lipopolysaccharide‑induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury. Experimental and Therapeutic Medicine, 30, 168. https://doi.org/10.3892/etm.2025.12918
MLA
Park, U., Kim, J."Yuanhuacine modulates lipopolysaccharide‑induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury". Experimental and Therapeutic Medicine 30.3 (2025): 168.
Chicago
Park, U., Kim, J."Yuanhuacine modulates lipopolysaccharide‑induced interleukin-6 through regulation of the JAK1/STAT3 pathway and prevents tubular damage in acute kidney injury". Experimental and Therapeutic Medicine 30, no. 3 (2025): 168. https://doi.org/10.3892/etm.2025.12918
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