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Baicalin mitigates polycystic ovary syndrome‑associated non‑alcoholic fatty liver disease by inhibiting the AR/SREBP1 axis

  • Authors:
    • Bi-Hui Jin
    • Han Xu
    • Zi-Yan Zhang
    • Yu-Hang Fan
    • Chun-Yan Jiang
    • Shao-Long Qi
    • Chen-Yu Xiao
    • Xiao-Hua Fu
    • Ling-Bo Qian
  • View Affiliations / Copyright

    Affiliations: Center for Reproductive Medicine, Department of Reproductive Endocrinology, Zhejiang Provincial People's Hospital (Affiliated People's Hospital), Hangzhou Medical College, Hangzhou, Zhejiang 310014, P.R. China, Department of Physiology, School of Basic Medical Sciences and Forensic Medicine, Hangzhou Medical College, Hangzhou, Zhejiang 310053, P.R. China, Department of Reproductive Endocrinology, Center for Reproductive Medicine, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 315300, P.R. China, Department of Vascular Surgery, China‑Japan Union Hospital of Jilin University, Changchun, Jilin 130031, P.R. China, Center for Reproductive Medicine, Department of Gynecology, Zhejiang Provincial People's Hospital (Affiliated People's Hospital), Hangzhou Medical College, Hangzhou, Zhejiang 310014, P.R. China
    Copyright: © Jin et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 189
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    Published online on: September 8, 2025
       https://doi.org/10.3892/ijmm.2025.5630
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Abstract

Polycystic ovary syndrome (PCOS) is a common endocrine disorder frequently associated with metabolic disturbances, such as non‑alcoholic fatty liver disease (NAFLD), driven by hyperandrogenism‑induced lipogenesis. Baicalin (BA), a flavonoid derived from Scutellaria baicalensis, exhibits therapeutic potential in the treatment of PCOS; however, the specific mechanisms against PCOS‑associated NAFLD remain unclear. In the present study, a PCOS mouse model was established via subcutaneous implantation of dihydrotestosterone. Model validation confirmed irregular estrous cycles, ovarian histopathological abnormalities and altered serum hormone levels. Treatment with BA markedly alleviated NAFLD‑associated metabolic abnormalities, including central obesity, dyslipidemia and hepatic steatosis. Moreover, liver transcriptomics indicated that BA modulated lipid metabolism primarily through sterol regulatory element‑binding protein 1 (SREBP1)‑mediated lipogenesis. Results of western blot analysis confirmed that BA suppressed hepatic protein expression of SREBP1 and its downstream lipogenic enzymes, fatty acid synthase and acetyl‑CoA carboxylase, indicating inhibition of hepatic lipogenesis. As androgen receptor (AR) functions as an upstream transcriptional regulator of SREBP1, network pharmacological analysis highlighted AR as a potential target of BA. Molecular docking predicted the BA‑AR binding site, guiding purification of truncated AR protein for isothermal titration calorimetry (ITC). Subsequently ITC was used to confirm the specific BA‑AR binding affinity. Luciferase reporter assays in MDA‑kb2 cells demonstrated that BA inhibited AR transcriptional activity. Collectively, the results of the present study indicated that BA ameliorates PCOS‑associated NAFLD through targeting the AR/SREBP1 axis, highlighting its potential as a therapeutic strategy for managing lipid metabolism disorders in PCOS.
View Figures

Figure 1

Validation of the PCOS mouse model.
(A) Chemical structure of BA. (B) The experimental workflow for
animal studies. (C) Representative images of vaginal exfoliated
cells in control mice at different estrous cycle stages. Proestrus,
predominantly nucleated epithelial cells (single or clustered) with
few leukocytes; estrus, large, anucleated, keratinized squamous
cells with irregular edges and minimal leukocyte presence;
metestrus, reduced keratinized epithelial cells accompanied by
numerous leukocytes and nucleated epithelial cells; diestrus,
dominated by leukocytes. (D) Representative estrous cycle patterns
in control and PCOS mice. (E) Representative ovarian sections
stained with H&E. Corpus luteum is indicated by & and
cystic follicles are indicated by #. Quantitative comparison of (F)
corpora lutea and (G) cystic follicles in control and PCOS groups.
Serum levels of (H) FSH, (I) LH, (J) DHT and (K) LH/FSH ratio in
control and PCOS groups. Data are presented as the mean ± standard
deviation (n=6). **P<0.01. Statistical significance
was determined using an unpaired twotailed Student's ttest. PCOS,
polycystic ovary syndrome; BA, baicalin; H&E, hematoxylin and
eosin; FSH, follicle-stimulating hormone; LH, luteinizing hormone;
DHT, dihydrotestosterone.

Figure 2

BA treatment alleviates
NAFLD-associated features in PCOS mice. (A) Body weight, (B)
visceral fat-to-body weight ratio, serum levels of (C) TG, (D) TC
and (E) NEFA in each group. (F) Representative liver histology in
each group. (G) Quantitative analysis of Oil Red O-positive areas
in liver sections. Hepatic levels of (H) TG, (I) TC and (J) NEFA in
each group. Data are presented as the mean ± standard deviation
(n=6-8). **P<0.01. Statistical significance was
assessed using oneway ANOVA followed by Tukey's post hoc test. BA,
baicalin; NAFLD, non-alcoholic fatty liver disease; PCOS,
polycystic ovary syndrome; TG, triglycerides; TC, total
cholesterol; NEFA, non-esterified fatty acids.

Figure 3

BA modulates lipid
metabolism-associated transcriptomic profiles in the liver of PCOS
mice. (A) PCA revealed distinct clustering of samples from control,
PCOS and BA treatment groups. Volcano plots depicted DEGs between
(B) PCOS vs. control and (C) BA-treated vs. PCOS groups. (D) Venn
diagram illustrated overlapping DEGs in each group. DEGs were
defined by DESeq2 with |log2(fold change)|>1.5 and
BH-adjusted q<0.05. (E) GO enrichment analysis of overlapping
DEGs highlighted the top 15 enriched biological processes
associated with lipid metabolism. (F) KEGG pathway enrichment
analysis of overlapping DEGs identified key pathways modulated by
BA. GO and KEGG enrichment was assessed using the default
hypergeometric test in clusterProfiler, followed by BH-adjusted
q<0.05. BA, baicalin; PCOS, polycystic ovary syndrome; PCA,
principal component analysis; DEGs, differentially expressed genes;
BH, Benjamini-Hochberg; GO, Gene Ontology; KEGG, Kyoto Encyclopedia
of Genes and Genomes.

Figure 4

BA regulates de novo
lipogenesis in PCOS mice through modulating the SREBP1 signaling
pathway. (A) Transcriptomic analysis of Srebp1 gene
expression levels in liver tissues (n=4). (B) Relative mRNA
expression of Srebp1 determined using qualitative PCR (n=8).
(C) Western blot analysis of SREBP1, ACC and FASN levels in the
liver tissue. Quantitative analysis of (D) SREBP1, (E) ACC and (F)
FASN protein expression levels. Data are presented as the mean ±
standard deviation. *P<0.05, **P<0.01.
Statistical significance was assessed using oneway ANOVA followed
by Tukey's post hoc test. BA, baicalin; PCOS, polycystic ovary
syndrome; Srebp1, sterol regulatory element-binding protein
1; ACC, acetyl-CoA carboxylase; FASN, fatty acid synthase.

Figure 5

Identification and analysis of
therapeutic targets of BA for PCOS-associated NAFLD. (A) BATMAN-TCM
identified potential drug targets of BA. GeneCards, OMIM and
DrugBank identified disease-associated targets for (B) PCOS and (C)
NAFLD. (D) Venn diagram illustrated the overlapping targets among
PCOS, NAFLD and BA. (E) PPI network of 25 common genes identified
in the Venn diagram. (F) CytoHubba analysis identified the top 10
core genes. BA, baicalin; PCOS, polycystic ovary syndrome; NAFLD,
non-alcoholic fatty liver disease; PPI, protein-protein
interaction.

Figure 6

Molecular docking, protein
purification and ITC analysis reveal binding between BA and AR.
Molecular docking analysis illustrated (A) 2D and (B) 3D binding
interactions between BA and AR. (C) Structural diagram of the
pGEX4T1-AR plasmid, with the sequence from the N-terminal to
C-terminal consisting of a GST tag, restriction sites and the
truncated AR protein. (D) GST-AR protein purification and Coomassie
Brilliant Blue staining for identification. The order of the
samples is indicated at the top of the image. The last lane
contains the purified GST-AR protein. ITC analysis of BA binding to
a truncated AR protein in (E) sample cells containing 1 μM
AR protein (GST-tagged) and in (F) sample cells containing 1
μM GST protein (control). Upper panels, heat flow vs. time;
lower panels, normalized heat effects vs. ligand-to-protein molar
ratio. ITC, isothermal titration calorimetry; BA, baicalin; AR,
androgen receptor; BSA, bovine serum albumin; GST, glutathione
S-transferase.

Figure 7

BA inhibits AR transcriptional
activity in MDA-kb2 cells. Results of the luciferase reporter assay
revealed (A) AR activation through increasing DHT concentrations
and (B) BA-mediated AR inhibition in the presence of 1 nM DHT in
MDA-kb2 cells. BA, baicalin; AR, androgen receptor; DHT,
dihydrotestosterone.

Figure 8

Proposed mechanism by which BA
ameliorates PCOS-associated NAFLD via the AR/SREBP1 signaling axis.
BA binds to AR and inhibits its transcriptional activity, thereby
downregulating the AR-mediated activation of SREBP1. This
suppression leads to decreased expression of key lipogenic enzymes,
including FASN and ACC, resulting in reduced hepatic lipogenesis.
Through this pathway, BA alleviates hepatic steatosis and metabolic
dysfunction in a DHT-induced mouse model of PCOS with NAFLD. BA,
baicalin; PCOS, polycystic ovary syndrome; NAFLD, non-alcoholic
fatty liver disease; AR, androgen receptor; SREBP1, sterol
regulatory element-binding protein 1; FASN, fatty acid synthase;
ACC, acetyl-CoA carboxylase; DHT, dihydrotestosterone.
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Copy and paste a formatted citation
Spandidos Publications style
Jin B, Xu H, Zhang Z, Fan Y, Jiang C, Qi S, Xiao C, Fu X and Qian L: Baicalin mitigates polycystic ovary syndrome‑associated non‑alcoholic fatty liver disease by inhibiting the AR/SREBP1 axis. Int J Mol Med 56: 189, 2025.
APA
Jin, B., Xu, H., Zhang, Z., Fan, Y., Jiang, C., Qi, S. ... Qian, L. (2025). Baicalin mitigates polycystic ovary syndrome‑associated non‑alcoholic fatty liver disease by inhibiting the AR/SREBP1 axis. International Journal of Molecular Medicine, 56, 189. https://doi.org/10.3892/ijmm.2025.5630
MLA
Jin, B., Xu, H., Zhang, Z., Fan, Y., Jiang, C., Qi, S., Xiao, C., Fu, X., Qian, L."Baicalin mitigates polycystic ovary syndrome‑associated non‑alcoholic fatty liver disease by inhibiting the AR/SREBP1 axis". International Journal of Molecular Medicine 56.5 (2025): 189.
Chicago
Jin, B., Xu, H., Zhang, Z., Fan, Y., Jiang, C., Qi, S., Xiao, C., Fu, X., Qian, L."Baicalin mitigates polycystic ovary syndrome‑associated non‑alcoholic fatty liver disease by inhibiting the AR/SREBP1 axis". International Journal of Molecular Medicine 56, no. 5 (2025): 189. https://doi.org/10.3892/ijmm.2025.5630
Copy and paste a formatted citation
x
Spandidos Publications style
Jin B, Xu H, Zhang Z, Fan Y, Jiang C, Qi S, Xiao C, Fu X and Qian L: Baicalin mitigates polycystic ovary syndrome‑associated non‑alcoholic fatty liver disease by inhibiting the AR/SREBP1 axis. Int J Mol Med 56: 189, 2025.
APA
Jin, B., Xu, H., Zhang, Z., Fan, Y., Jiang, C., Qi, S. ... Qian, L. (2025). Baicalin mitigates polycystic ovary syndrome‑associated non‑alcoholic fatty liver disease by inhibiting the AR/SREBP1 axis. International Journal of Molecular Medicine, 56, 189. https://doi.org/10.3892/ijmm.2025.5630
MLA
Jin, B., Xu, H., Zhang, Z., Fan, Y., Jiang, C., Qi, S., Xiao, C., Fu, X., Qian, L."Baicalin mitigates polycystic ovary syndrome‑associated non‑alcoholic fatty liver disease by inhibiting the AR/SREBP1 axis". International Journal of Molecular Medicine 56.5 (2025): 189.
Chicago
Jin, B., Xu, H., Zhang, Z., Fan, Y., Jiang, C., Qi, S., Xiao, C., Fu, X., Qian, L."Baicalin mitigates polycystic ovary syndrome‑associated non‑alcoholic fatty liver disease by inhibiting the AR/SREBP1 axis". International Journal of Molecular Medicine 56, no. 5 (2025): 189. https://doi.org/10.3892/ijmm.2025.5630
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