Transcriptomic characteristics of cervical endplate cartilage in cervical spondylosis and effects of acupotomy on the FGF18/Akt axis with cervical spondylosis

doi:10.12122/j.issn.1673-4254.2026.01.06

Abstract

Abstract:

Objective To explore the transcriptomic characteristics of the endplate cartilage of the cervical intervertebral disc in cervical spondylosis (CS) and the effects of acupotomy on expressions of fibroblast growth factor 18 (FGF18)/protein kinase B (Akt) axis key molecules. Methods Transcriptomic analyses were performed using bioinformatics methods based on the GEO database. In the animal experiment, 24 New Zealand rabbits were randomized equally into control, CS model, and acupotomy groups. In the latter two groups, CS models were established followed 7 days later by acupotomy intervention (once a week for 3 weeks) or no particular treatment. Histopathological changes and cell apoptosis in the intervertebral discs were examined with HE staining and TUNEL assay, and the mRNA expressions of FGF18, FGFR3, and Akt in the intervertebral discs were detected using RT-qPCR; the localization and expressions of FGF18, p-Akt, and Akt proteins were examined with immunohistochemistry. Results The endplate cartilage in CS exhibited numerous differentially expressed genes enriched in the PI3K-Akt signaling, calcium signaling, and Rap1 signaling pathways. CS rabbits showed obvious changes in the cervical spine curvature and joint degenerations, changes in cervical intervertebral disc texture, thinning of the annulus fibrosus, shrinkage or even absence of the nucleus pulposus, and increased apoptotic cells in the endplate cartilage, which were all obviously alleviated after acupotomy. No significant differences were found in the mRNA expressions of FGF18, FGFR3, and Akt in the cervical intervertebral discs among the 3 groups. Acupotomy significantly increased FGF18 and p-Akt protein expressions and reduced the Akt/p-Akt ratio in the cervical endplate cartilage of CS rabbits. Conclusion The cervical endplate cartilage of CS show numerous differentially expressed genes. Acupotomy may delay degenerative changes of the intervertebral discs and improves CS by activating the FGF18/Akt axis to reduce apoptosis of endplate cartilage cells.

Key words: cervical spondylosis, acupotomy, transcriptome characteristics, intervertebral disc, FGF18/Akt axis

Fushui LIU, Tumurbaatar KHALIUNAA, Qiguang CAO, Yuqian YANG, Changan REN, Jinchao ZHU, Xiaolan ZHAO, Li CAO, Biao DENG, Xiaole WANG. Transcriptomic characteristics of cervical endplate cartilage in cervical spondylosis and effects of acupotomy on the FGF18/Akt axis with cervical spondylosis[J]. Journal of Southern Medical University, 2026, 46(1): 55-65.

Figures/Tables 12

Fig.1 Construction of cervical spondylosis (CS) rabbit model.

Fig.2 Palpation areas selected as the treatment points.

Tab.1 Primer sequences for RT-qPCR in this study

Gene	Primer sequence (5'-3')
GAPDH-sense	TTCCAGTATGATTCCACCCACG
GAPDH-antisense	GGGCTGAGATGATGACCCTTTT
FGF18-sense	GAGTGTGTGTTCATCGAGAAGGTTC
FGF18-antisense	GAGCGCTTGGTCACTGTGGT
FGFR3-sense	AGGAACAGGTGGTCTTTGGCA
FGFR3-antisense	CGCACAATGAAGTGGCACAG
AKT1-sense	AGTACATCAAGACCTGGCGG
AKT1-antisense	AGTTGTTGAGTGGGGACTCG

Fig.3 Analysis of differentially expressed genes. A: Volcano plot of the differentially expressed genes (Blue, red, and gray dots represent genes with decreased expression, increased expression, and no significantly changes expression, respectively). B: Heatmap of the differentially expressed genes.

Fig.4 Analysis of the functions of differentially expressed genes. A: Top 10 enriched pathways and the related differentially expressed genes (DEGs). B: GSEA analysis of the top 10 enriched pathways. C: Heatmap of the DEGs in the top 10 enriched pathways. D: Expression of FGF18 mRNA. n=3, #P<0.05.

Fig.5 Changes in X-ray of rabbit cervical vertebrae before and after modeling. A: X-ray before modeling. B: X-ray after modeling.

Fig.6 HE staining of cervical intervertebral discs of the rabbits in each group. A: Control group. B: CS group. C: Acupotomy group.

Fig.7 FGF18, FGFR3, and Akt1 mRNA expressions in the cervical intervertebral discs of the rabbits in each group. n=6.

Fig.8 Expression of FGF18 protein in the nucleus pulposus, annulus fibrosus, and endplate cartilage of the cervical intervertebral discs of the rabbits in each group. n=6, #P<0.05.

Fig.9 Expression of Akt protein in the nucleus pulposus, annulus fibrosus, and endplate cartilage of the cervical intervertebral discs of rabbits from each group. n=6, #P<0.05.

Fig.10 Expression of p-Akt protein in the nucleus pulposus, annulus fibrosus, and endplate cartilage and the Akt to p-Akt ratio in the cervical intervertebral discs of the rabbits in each group. n=6, #P<0.05.

Fig.11 Apoptosis of endplate cartilage cells in the cervical intervertebral discs of the rabbits in each group. n=3, #P<0.05.

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