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Lactate regulates osteoclastogenesis via H3k18la in osteoarthritis

  • Authors:
    • Zhibo Fan
    • Qiyue Fu
    • Tao Sun
    • Yutong Zhong
    • Yuefan Ma
    • Jingwei Jian
    • Shuosheng Yuan
    • Shenghong Li
    • Xiaomei Xu
  • View Affiliations / Copyright

    Affiliations: Department of Orthodontics, The Affiliated Stomatological Hospital of Southwest Medical University, Luzhou, Sichuan 646000, P.R. China
    Copyright: © Fan et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 4.0].
  • Article Number: 207
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    Published online on: June 2, 2026
       https://doi.org/10.3892/ijmm.2026.5878
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Abstract

Osteoarthritis (OA), characterized by articular cartilage degeneration and subchondral bone remodeling, is a notable cause of disability globally. The present study aimed to explore the role and mechanism of histone H3 lysine 18 lactylation (H3K18la) in OA osteoclast differentiation and verify the therapeutic potential of targeting this pathway. Using a mouse OA model (anterior cruciate ligament transection), RAW264.7 cell experiments, oxmacid‑mediated lactate inhibition and cleavage under targets and tagmentation to map genomic targets, the present study demonstrated elevated H3K18la in OA joint osteoclasts, associated with reduced bone mineral density and aggravated subchondral bone destruction. Analysis of TRAP and lactate indicated that RANKL‑induced osteoclast differentiation increased lactate production, enhancing H3K18la; oxmacid inhibited both of these processes. H3K18la was enriched at the acid phosphatase 5 promoter and directly promoted its transcription. Local oxmacid injection in OA mice decreased osteoclast numbers and alleviated subchondral bone loss. Thus, H3K18la was a key metabolic‑epigenetic mediator linking glycolysis to osteoclast differentiation, representing a novel OA therapeutic target.
View Figures

Figure 1

Analysis of the RANKL-treated and
control groups based on RNA-sequencing data. (A) PCA plot. (B)
Volcano plot. (C) Kyoto Encyclopedia of Genes and Genomes
enrichment analysis based on the significantly differentially
expressed genes (P<0.01 and |log2FC| >1). (D)
Bubble plot of pathway enrichment analysis (P<0.01 and
|log2FC| >1), focusing on carbohydrate
metabolism-related pathways. (n=5). PCA, principal component
analysis; KEGG; FC, fold-change.

Figure 2

H3K18la increases during osteoclastic
differentiation. (A) TRAP staining of macrophages following RANKL
(40 ng/ml) treatment. (B) TRAP quantitative analysis of
macrophages. (C) Relative protein expression of Pan-Kla, H3K18la
and LDHA. (D) Lactate quantitative analysis of macrophages. (E)
Immunofluorescence staining of H3K18la and F-actin protein in the
knee. n=3. *P<0.05; **P<0.01;
***P<0.001 vs. day 0. H3K18la, histone H3 lactylation
at lysine 18; TRAP, tartrate-resistant acid phosphatase; LDHA,
lactate dehydrogenase A; Pan-Kla, pan-lysine lactylation; MNC,
Multinucleated Cells; prot, protein.

Figure 3

Elevated H3K18la in osteoclasts of OA
model mice. (A) TRAP staining of the knees of mice 2 weeks after
surgery. Arrows indicate osteoclasts. (B) SO/FG staining of tissue
from the knee. (C) Representative micro-CT 3D images of the knee.
(D) Multiple-immunofluorescence staining of Pan-kla, H3K18la and
Ctsk proteins in the knee tissue. Arrows indicate areas of
co-localization. n=3. *P<0.05;
**P<0.01; ***P<0.001. H3K18la, histone
H3 lactylation at lysine 18; OA, osteoarthritis; TRAP,
tartrate-resistant acid phosphatase; SO/FG, Safranin O/fast green;
Ctsk, Cathepsin K; ACLT, anterior cruciate ligament transection;
ROI, region of interest.

Figure 4

Inhibition of H3K18la suppresses
osteoclastic differentiation. (A) Experimental inhibition of
macrophage osteoclast differentiation. (B) Toxicity, (C) lactate
levels and (D) TRAP-positive staining of macrophages treated with
lactate inhibitors (0, 5, 10 and 20 mM). (E) TRAP staining, (F)
relative protein expression of Pan-Kla, H3K18la and LDHA and (G)
immunofluorescence staining of H3K18la and F-actin protein
following treatment with lactate inhibitors (0, 5, 10 and 20 mM).
n=3. *P<0.05; **P<0.01;
***P<0.001 vs. 0 mM. H3K18la, histone H3 lactylation
at lysine 18; TRAP, tartrate-resistant acid phosphatase; Ctsk,
Cathepsin K; LDHA, lactate dehydrogenase A; prot, protein; CCK-8,
Cell Counting Kit-8; OD, optical density; Pan-Kla, Pan-lysine
lactylation; MNC, multinucleated cells.

Figure 5

Inhibition of histone lactylation
alleviates OA pathology. (A) Hematoxylin and eosin staining of the
heart, liver, spleen, lung and kidney of mice. (B)
Immunofluorescence staining of the H3K18la protein in the knee. (C)
Representative micro-CT 3D of the knee. (D) TRAP staining of the
knees of mice with or without lactate inhibitors. Mice received
continuous local injection of oxamic acid in the knees at 2 weeks
after the operation. Arrows indicate osteoclasts. (E) SO/FG
staining of the knee. Black arrows indicate cartilage
discontinuity; triangle indicates subchondral bone resorption.
(n=3). *P<0.05; **P<0.01;
***P<0.001 vs. ACLT. OA, osteoarthritis; H3K18la,
histone H3 lactylation at lysine 18; TRAP, tartrate-resistant acid
phosphatase; SO/FG, Safranin O/fast green; ACLT, anterior cruciate
ligament transection.

Figure 6

Identification of H3K18la targets
during osteoclastic differentiation. (A) Binding density of H3K18la
in macrophages with and without RANKL treatment. (B) KEGG
enrichment analysis of genes with differential H3K18la binding
(increased in RANKL-treated vs. control) (n=1). (C) Genome-wide
distribution of H3K18la-binding peaks. (D) Gene Ontology enrichment
analysis of the differentially expressed genes (P<0.01 and
|log2FC| >1) between the two groups with RNA-seq
(P<0.05; n=5). (E) CUT&Tag and scRNA-seq analyses identified
upregulated targets of H3K18la (P<0.05, |log2FC|
>2.5). **P<0.01; ***P<0.001.
H3K18la, histone H3 lactylation at lysine 18; KEGG, Kyoto
Encyclopedia of Genes and Genomes; FC, fold-change; scRNA-seq,
single cell RNA sequencing; BP, biological process; CC, cellular
component; MF, molecular function; Nfatc1, nuclear factor of
activated T cells 1; Acp5, acid phosphatase 5; Ctsk, cathepsin K;
RPKM, reads per kilobase of transcript per million mapped reads;
Fosl2, FOS-like antigen 2.

Figure 7

H3K18la activates Acp5 transcription.
(A) Genome browser tracks of the CUT&Tag signal at NFATc1,
Acp5, Ctsk and Fosl2. Relative (B) mRNA expression of Nfatc1, Acp5,
Ctsk and Fosl2 and (C) protein expression of Pan-Kla, H3K18la and
Acp5 in macrophages with or without lactate (10 mM) treatment. (D)
TRAP staining of macrophages with or without lactate (10 mM)
treatment. (E) H3K18la binding at the Acp5 promoter assessed by
ChIP-qPCR. **P<0.01; ***P<0.001 vs.
controls. H3K18la, histone H3 lactylation at lysine 18; Acp5,
tartrate-resistant acid phosphatase type 5; NFATc1, nuclear factor
of activated T cells 1; Ctsk, Cathepsin K; Fosl2, FOS-like antigen
2; TRAP, tartrate-resistant acid phosphatase; CHIP-q, chromatin
immunoprecipitation-quantitative; Pan-Kla, Pan-lysine Lactylation;
TSS, transcription start site; chr, chromosome; ns, not
significant.

Figure 8

H3K18la affects osteoclast function
by regulating Acp5 expression. Inhibition of H3K18la alleviates
early bone destruction in osteoarthritis. H3K18la, histone H3
lactylation at lysine 18; Acp5, tartrate-resistant acid phosphatase
type 5; Lac, lactate.
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Copy and paste a formatted citation
Spandidos Publications style
Fan Z, Fu Q, Sun T, Zhong Y, Ma Y, Jian J, Yuan S, Li S and Xu X: Lactate regulates osteoclastogenesis via H3k18la in osteoarthritis. Int J Mol Med 58: 207, 2026.
APA
Fan, Z., Fu, Q., Sun, T., Zhong, Y., Ma, Y., Jian, J. ... Xu, X. (2026). Lactate regulates osteoclastogenesis via H3k18la in osteoarthritis. International Journal of Molecular Medicine, 58, 207. https://doi.org/10.3892/ijmm.2026.5878
MLA
Fan, Z., Fu, Q., Sun, T., Zhong, Y., Ma, Y., Jian, J., Yuan, S., Li, S., Xu, X."Lactate regulates osteoclastogenesis via H3k18la in osteoarthritis". International Journal of Molecular Medicine 58.2 (2026): 207.
Chicago
Fan, Z., Fu, Q., Sun, T., Zhong, Y., Ma, Y., Jian, J., Yuan, S., Li, S., Xu, X."Lactate regulates osteoclastogenesis via H3k18la in osteoarthritis". International Journal of Molecular Medicine 58, no. 2 (2026): 207. https://doi.org/10.3892/ijmm.2026.5878
Copy and paste a formatted citation
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Spandidos Publications style
Fan Z, Fu Q, Sun T, Zhong Y, Ma Y, Jian J, Yuan S, Li S and Xu X: Lactate regulates osteoclastogenesis via H3k18la in osteoarthritis. Int J Mol Med 58: 207, 2026.
APA
Fan, Z., Fu, Q., Sun, T., Zhong, Y., Ma, Y., Jian, J. ... Xu, X. (2026). Lactate regulates osteoclastogenesis via H3k18la in osteoarthritis. International Journal of Molecular Medicine, 58, 207. https://doi.org/10.3892/ijmm.2026.5878
MLA
Fan, Z., Fu, Q., Sun, T., Zhong, Y., Ma, Y., Jian, J., Yuan, S., Li, S., Xu, X."Lactate regulates osteoclastogenesis via H3k18la in osteoarthritis". International Journal of Molecular Medicine 58.2 (2026): 207.
Chicago
Fan, Z., Fu, Q., Sun, T., Zhong, Y., Ma, Y., Jian, J., Yuan, S., Li, S., Xu, X."Lactate regulates osteoclastogenesis via H3k18la in osteoarthritis". International Journal of Molecular Medicine 58, no. 2 (2026): 207. https://doi.org/10.3892/ijmm.2026.5878
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