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Article Open Access

Integrated network pharmacology and serum metabolomics to reveal the protective mechanism of methanolic extract of BaiYangJie on cisplatin‑induced acute kidney injury in mice

  • Authors:
    • Bin Xia
    • Jianglong Chen
    • Jinhui Wang
    • Dalong Li
    • Lixia Zhang
    • Jing Su
    • Xuan Ding
    • Yana Lv
    • Shifang Liu
    • Xue Zhang
    • Tianzhen Wan
    • Guang Li
  • View Affiliations / Copyright

    Affiliations: School of Pharmacy, Heilongjiang University of Chinese Medicine, Haerbin, Heilongjiang 150006, P.R. China, Pharmacology Center, Chinese Academy of Medical Sciences and Peking Union Medical College Institute of Medicinal Plant Development Yunnan Branch, Jinghong, Yunnan 666100, P.R. China, Key Laboratory of Biology and Genetic Improvement of Horticulture Crops (Northeast Region), Ministry of Agriculture and Rural Affairs, School of Horticulture and Landscape Architecture, Northeast Agricultural University, Harbin, Heilongjiang, 150006, P.R. China
    Copyright: © Xia et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 205
    |
    Published online on: June 4, 2026
       https://doi.org/10.3892/etm.2026.13199
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Abstract

Cisplatin, a chemotherapeutic drug, produces severe nephrotoxicity and at present, there are no effective drugs to clinically prevent or treat it. In the Dai nationality, BaiYangJie is used to treat poisoning caused by chemicals or drugs. Previous research has demonstrated that the methanolic extract of BaiYangJie (MEAG) can treat cisplatin‑induced nephrotoxicity; however, its mechanism of action remains unclear. The aim of the present study was therefore to identify the potential mechanism of action of MEAG in cisplatin nephrotoxicity using a combination of network pharmacology and serum metabolomics. Initially, network pharmacology analysis was used to identify hub targets and signalling pathways involved in the renoprotective effects of MEAG. Subsequently, plasma metabolomics profiling utilising ultra performance liquid chromatography‑quadrupole time‑of‑flight mass spectrometry technology revealed key metabolic alterations and pathway modulations associated with MEAG treatment. Finally, integrated analysis uncovered key molecular mechanisms, which were subsequently validated by western blotting and immunohistochemistry. A total of 13 endogenous metabolites were identified in serum metabolomics, primarily involved in phenylalanine metabolism and in the biosynthesis of phenylalanine, tyrosine and tryptophan. The treatment of MEAG in cis‑induced acute kidney injury primarily involved regulating the inflammatory response, responses to lipid and chemical stress, the FoxO signalling pathway, arachidonic acid metabolism and the NF‑κB signalling pathway. Animal experiments showed that MEAG can inhibit inflammation and the expression of the migration inhibitory factor (MIF)/NF‑κB pathway. Through integrated network pharmacological analysis, metabolomic profiling and experimental validation, it was systematically elucidated that MEAG exerted its therapeutic effects through dual regulatory mechanisms: Suppressing inflammatory responses by inhibiting overactivation of the MIF/NF‑κB signalling pathway and restoring phenylalanine metabolic homeostasis. These findings thus provide a mechanistic foundation for developing targeted therapeutic strategies against chemotherapy‑associated nephrotoxicity.
View Figures

Figure 1

Base peak chromatogram of methanolic
extract of BaiYangJie. (A) Positive ion modes and (B) negative ion
modes.

Figure 2

Therapeutic mechanisms of MEAG
against Cis-AKI in murine models using network pharmacology
approaches. (A) Venn analysis of common targets of MEAG-Cis-AKI in
mice. (B) Protein-protein interaction network of shared MEAG and
Cis-AKI targets. (C) Core protein-protein interaction network for
MEAG against Cis-AKl injury. (D) Bar diagram of top 20 biological
process, cellular component and molecular function terms. (E)
Enrichment profile of the 20 most significant Kyoto Encyclopaedia
of Genes and Genomes signalling pathways. MEAG, methanolic extract
of BaiYangJie; Cis, cisplatin; AKI, acute kidney injury.

Figure 3

Multivariate statistical analyses of
metabolomics in mice serum. (A) PCA score plot of serum metabolism
in POS ionization mode. (B) PCA score plot of serum metabolism in
NEG ionization mode. (C) OPLS-DA score plot comparing the Nor and
Cis groups in POS ionization mode. (D) S-plot comparing the Nor and
Cis groups in POS ionization mode. (E) OPLS-DA score plot comparing
the Nor and Cis groups in NEG ionization mode. (F) S-plot comparing
the Nor and Cis groups in NEG ionization mode. (G) OPLS-DA score
plot comparing the Cis and MEAG groups in POS ionization mode. (H)
S-plot comparing the Cis and MEAG groups in POS ionization mode.
(I) OPLS-DA score plot comparing the Cis and MEAG groups in NEG
ionization mode. (J) S-plot comparing the Cis and MEAG groups in
NEG ionization mode. OPLS-DA, orthogonal partial least squares
discriminant analysis; Nor, normal control group; Cis, cisplatin;
MEAG, methanolic extract of BaiYangJie; QC, quality control; POS,
positive; NEG, negative; PCA, principal component analysis.

Figure 4

Cluster thermogram of all endogenous
differential metabolites. Nor, normal control group; Cis,
cisplatin; MEAG, methanolic extract of BaiYangJie; LysoPE,
lysophosphatidylethanolamine.

Figure 5

Relative content changes of potential
metabolites. ####P<0.0001, ##P<0.01,
#P<0.05; compared with the Nor group,
****P<0.0001, ***P<0.001,
**P<0.01 and *P<0.05 compared with the
Cis group. LysoPE, lysophosphatidylethanolamine; POS, positive;
NEG, negative; Cis, cisplatin; MEAG, methanolic extract of
BaiYangJie; Nor, normal control group.

Figure 6

Metabolic pathway analysis. (A)
Metabolic modules regulated by the methanolic extract of
BaiYangJie. (B) Association diagram of ‘phenylalanine metabolism’
and ‘phenylalanine, tyrosine and tryptophan biosynthesis’.

Figure 7

Schematic diagram of MEAG regulating
phenylalanine metabolic flux. MEAG, methanolic extract of
BaiYangJie; Cis, cisplatin; Cis-AKI, cisplatin-induced acute kidney
injury; Phe, phenylalanine; Tyr, tyrosine; MIF, macrophage
migration inhibitory factor; NF-κB, nuclear factor κB; P-NF-κB,
phosphorylated NF-κB; TNF-α, tumor necrosis factor α; IL-6,
interleukin-6; IL-1β, interleukin-1β; AKI, acute kidney injury.

Figure 8

MEAG treatment ameliorated Cis-AKI in
mice. (A) Effect of MEAG on body weight change in mice with
Cis-AKI. (B) Effect of MEAG on kidney index in mice with Cis-AKI.
(C) Effect of MEAG on BUN levels in mice with Cis-AKI. (D) Effect
of MEAG on Scr levels in mice with Cis-AKI. (E) Representative
H&E-stained kidney sections from the Nor, Cis, and MEAG groups,
shown at x200 magnification with corresponding higher-magnification
views at x400. Yellow arrows indicate rubber-like tubular casts,
blue arrows indicate shedding of renal tubular epithelial cells,
green arrows indicate renal tubular dilatation, and black arrows
indicate glomerular atrophy and basement membrane thickening. (F)
Tubular injury score in each group. (G) Representative
Masson-stained kidney sections from the Nor, Cis and MEAG groups,
shown at x200 magnification with corresponding higher-magnification
views at x400. (H) Collagen volume fraction in each group. Data in
panels (A-D), (F) and (H) are presented as mean ± SD (n=6 mice per
group). ####P<0.0001 compared with the Nor group;
**P<0.01, ***P<0.001 and
****P<0.0001 compared with the Cis group. Scale bars,
50 µm for x200 images and 20 µm for x400 images. Nor, normal
control group; Cis, cisplatin; MEAG, methanolic extract of
BaiYangJie; AKI, acute kidney injury; Scr, serum creatinine; BUN,
blood urea nitrogen.

Figure 9

Upregulation of KIM-1 and NGAL
expression induced by cisplatin was reduced by MEAG. (A)
Representative IHC images of KIM-1 expression in kidney tissues
from the Nor, Cis and MEAG groups. (B) Representative IHC images of
NGAL expression in kidney tissues from the Nor, Cis and MEAG
groups. (C) Quantitative analysis of the positive stained area of
KIM-1 in kidney tissues. (D) Quantitative analysis of the positive
stained area of NGAL in kidney tissues. For panels (A) and (B),
magnification x200 and scale bar, 50 µm. For panels (C) and (D),
data are presented as mean ± SD (n=3 per group).
####P<0.0001, compared with the Nor group and
****P<0.0001, compared with the Cis group. Nor,
normal control group; Cis, cisplatin; MEAG, methanolic extract of
BaiYangJie; IHC, immunohistochemistry; NGAL, neutrophil
gelatinase-associated lipocalin; KIM-1, kidney injury molecule
1.

Figure 10

MEAG alleviated Cis-AKI in mice by
regulating the expression of MIF/NF-κB. (A-C) Serum concentrations
of pro-inflammatory cytokines (A) IL-1β, (B) IL-6 and (C) TNF-α
were quantified through ELISA across experimental cohorts
(n=3/group). (D) Renal tissue MIF distribution patterns visualized
through IHC staining (n=3/group). (E) Semiquantitative evaluation
of MIF immunoreactivity intensity. (F) Western blotting profiles
demonstrating protein expression alterations in MIF, p-NF-κB,
IL-1β, IL-6 and TNF-α protein levels. (G-J) Densitometric analysis
of (G) MIF, (H) IL-1β, (I) IL-6 and (J) TNF-α (n=3/group). (K)
Western blotting profiles demonstrating protein expression
alterations in p-NF-κB and NF-κB. (L) Densitometric analysis of
p-NF-κB/NF-κB (n=3/group). ####P<0.0001,
###P<0.001, compared with the Nor group;
****P<0.0001, ***P<0.001 and
**P<0.01, compared with the Cis group. Magnification,
x200 and scale bar, 50 µm. Nor, normal control group; Cis,
cisplatin; MEAG, methanolic extract of BaiYangJie; AKI, acute
kidney injury; IHC, immunohistochemistry; MIF, migration inhibitory
factor; p-, phosphorylated.
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Copy and paste a formatted citation
Spandidos Publications style
Xia B, Chen J, Wang J, Li D, Zhang L, Su J, Ding X, Lv Y, Liu S, Zhang X, Zhang X, et al: Integrated network pharmacology and serum metabolomics to reveal the protective mechanism of methanolic extract of BaiYangJie on cisplatin‑induced acute kidney injury in mice. Exp Ther Med 32: 205, 2026.
APA
Xia, B., Chen, J., Wang, J., Li, D., Zhang, L., Su, J. ... Li, G. (2026). Integrated network pharmacology and serum metabolomics to reveal the protective mechanism of methanolic extract of BaiYangJie on cisplatin‑induced acute kidney injury in mice. Experimental and Therapeutic Medicine, 32, 205. https://doi.org/10.3892/etm.2026.13199
MLA
Xia, B., Chen, J., Wang, J., Li, D., Zhang, L., Su, J., Ding, X., Lv, Y., Liu, S., Zhang, X., Wan, T., Li, G."Integrated network pharmacology and serum metabolomics to reveal the protective mechanism of methanolic extract of BaiYangJie on cisplatin‑induced acute kidney injury in mice". Experimental and Therapeutic Medicine 32.2 (2026): 205.
Chicago
Xia, B., Chen, J., Wang, J., Li, D., Zhang, L., Su, J., Ding, X., Lv, Y., Liu, S., Zhang, X., Wan, T., Li, G."Integrated network pharmacology and serum metabolomics to reveal the protective mechanism of methanolic extract of BaiYangJie on cisplatin‑induced acute kidney injury in mice". Experimental and Therapeutic Medicine 32, no. 2 (2026): 205. https://doi.org/10.3892/etm.2026.13199
Copy and paste a formatted citation
x
Spandidos Publications style
Xia B, Chen J, Wang J, Li D, Zhang L, Su J, Ding X, Lv Y, Liu S, Zhang X, Zhang X, et al: Integrated network pharmacology and serum metabolomics to reveal the protective mechanism of methanolic extract of BaiYangJie on cisplatin‑induced acute kidney injury in mice. Exp Ther Med 32: 205, 2026.
APA
Xia, B., Chen, J., Wang, J., Li, D., Zhang, L., Su, J. ... Li, G. (2026). Integrated network pharmacology and serum metabolomics to reveal the protective mechanism of methanolic extract of BaiYangJie on cisplatin‑induced acute kidney injury in mice. Experimental and Therapeutic Medicine, 32, 205. https://doi.org/10.3892/etm.2026.13199
MLA
Xia, B., Chen, J., Wang, J., Li, D., Zhang, L., Su, J., Ding, X., Lv, Y., Liu, S., Zhang, X., Wan, T., Li, G."Integrated network pharmacology and serum metabolomics to reveal the protective mechanism of methanolic extract of BaiYangJie on cisplatin‑induced acute kidney injury in mice". Experimental and Therapeutic Medicine 32.2 (2026): 205.
Chicago
Xia, B., Chen, J., Wang, J., Li, D., Zhang, L., Su, J., Ding, X., Lv, Y., Liu, S., Zhang, X., Wan, T., Li, G."Integrated network pharmacology and serum metabolomics to reveal the protective mechanism of methanolic extract of BaiYangJie on cisplatin‑induced acute kidney injury in mice". Experimental and Therapeutic Medicine 32, no. 2 (2026): 205. https://doi.org/10.3892/etm.2026.13199
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