Abstract: BACKGROUND: The regulatory mechanisms of acupotomy intervention for knee osteoarthritis at a transcriptome level are not well understood despite its proven clinical efficacy.
OBJECTIVE: Using acupotomy therapy in a rat model of knee osteoarthritis, to conduct transcriptome sequencing and bioinformatics analysis on cartilage samples, along with validation, and to reveal the molecular regulatory mechanisms involved in this therapy for knee osteoarthritis in rats.
METHODS: Forty-eight Sprague-Dawley rats were selected and divided into three groups by random number table method, acupotomy group, model group, and sham operation group, with 16 rats in each group. Osteoarthritis models were induced by medial meniscus instability in the acupotomy group and model group. After successful modeling, acupotomy group rats were treated with acupotomy once a week, for 4 weeks in total. After the intervention, cartilage samples from the rat’s knee joint were stained with hematoxylin-eosin staining and safranin O-fast green staining, evaluated by Mankin scores, and underwent MicroCT scanning. Serum inflammatory factor levels were detected by Elisa. Transcriptome sequencing was performed on the remaining cartilage samples, and the data were analyzed using R software to identify differential gene expression levels among the groups. Core targets were screened through protein-protein interaction network and Cytoscape and validated using RT-qPCR.
RESULTS AND CONCLUSION: Compared with the sham operation group, rats in the model group had rough and uneven articular cartilage surfaces, narrowed joint spaces, destroyed articular surface structures, elevated Mankin scores, and significant increases in serum levels of interleukin-6, tumor necrosis factor-α, and matrix metalloproteinase 13 (P < 0.05). Compared with the model group, rats in the acupotomy group had smoother articular cartilage surfaces, wider joint spaces, slightly irregular articular surfaces, lower Mankin scores, and significantly lower serum levels of interleukin-6, tumor necrosis factor-α, and matrix metalloproteinase-13 (P < 0.05). Gene ontology and Kyoto genome and genome encyclopedia analyses involved proteolytic metabolism, autophagy, mitogen-activated protein kinase, nuclear factor kB, and Wnt signaling pathways. Protein-protein interaction network and Cytoscape screened for four key genes, including ataxia-telangiectasia mutations, Myb SWIRM and MPN domain protein 1, heat shock protein 90α1, and NIPBL cohesion-loading factor. The mRNA expression of ataxia-telangiectasia mutations, Myb SWIRM and MPN domain protein 1, heat shock protein 90α1, and NIPBL cohesion-loading factor in the articular cartilage of rats in the model group was lower than that of the sham operation group (P < 0.05), while the mRNA expression of ataxia-telangiectasia mutations, Myb SWIRM and MPN domain protein 1, heat shock protein 90α1, and NIPBL cohesion-loading factor in the articular cartilage of rats in the acupotomy group was higher than that in the model group (P < 0.05). To conclude, acupotomy treatment of knee osteoarthritis in rats may act on signaling pathways such as MAPK, nuclear factor kB, and Wnt to promote cartilage repair, and is closely related to the expression of genes associated with ataxia-telangiectasia mutations, Myb SWIRM and MPN domain protein 1, heat shock protein 90α1, and NIPBL cohesion-loading factor.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: knee osteoarthritis, acupotomy, osteoarthritis model, destabilization of the medial meniscus, transcriptome sequencing, differential genes, animal experiment, experimental verification, engineered tissue construction