Acupotomy can delay the progression of knee osteoarthritis (KOA), but its therapeutic mechanism is still unclear.

To investigate the therapeutic effect of acupotomy on KOA and related mechanism.

From May 2023 to February 2024, 40 healthy specific pathogen-free (SPF) New Zealand rabbits were randomly divided into blank group, model group, acupotomy group, acupotomy combined with A3 adenosine receptor (A3AR) antagonist group and acupotomy combined with β-catenin activator group by SPSS 26.0 software, with eight rabbits in each group. The rabbits in the blank group did not receive any intervention, and the rabbits in the other groups were injected with papain into the joint cavity, combined with forced flexion of the knee joint, to construct KOA models. The rabbits in the model group were not given any intervention after the model was established; rabbits in the acupotomy group were treated with acupotomy; rabbits in the acupotomy combined with A3AR antagonist group were treated with acupotomy and injection of A3AR antagonist MRS1220 (5 mg/kg) via the auricular vein; rabbits in the acupotomy combined with β-catenin activator group were treated with acupotomy and combined with injection of Wnt/β-catenin signaling pathway activator SKL2001 (5 mg/kg) through the auricular vein, once a week for 4 weeks. Before and after the intervention, the behaviors of rabbits in each group were observed. After the intervention, the knee joint of rabbits in each group was imaged, and then the gross morphological changes in the knee joint cartilage were observed. The pathological changes in the cartilage tissue were observed by hematoxylin-eosin (HE) staining. The Lequesne MG score, Kellgren-Lawrence (K-L) grade, Pelletier score and Mankin score were used for quantitative evaluation. The content of pro-inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-18 (IL-18), and matrix metalloproteinase-13 (MMP-13) in knee cartilage tissue and joint fluid were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA levels of A3AR, Wnt3a, β-catenin, Nod-like receptor protein 3 (NLRP3), Caspase-1 and Gasdermin-D (GSDMD) in knee cartilage tissue were detected by real-time polymerase chain reaction (Real-time PCR), and the relative protein expression levels of A3AR, Wnt3a, β-catenin, NLRP3, Caspase-1 and GSDMD in knee cartilage tissue were detected by Western Blotting.

Compared with the blank group, the model group showed obvious knee joint degeneration, and higher the Lequesne MG score, K-L grade, Pelletier score and Mankin score (P<0.05), contents of TNF-α, IL-1β, IL-18, and MMP-13 in knee cartilage tissue and joint fluid (P<0.05), lower mRNA levels and protein relative expressions in A3AR of knee cartilage tissue (P<0.05), and higher mRNA levels and protein relative expressions of Wnt3a, β-catenin, NLRP3, Caspase-1 and GSDMD in knee cartilage tissue (P<0.05). Compared with the model group, the above changes in the acupotomy group were reversed, and tended to return to the normal range (P<0.05). Compared with the acupotomy group, both the acupotomy combined with A3AR antagonist group and the acupotomy combined with β-catenin activator group showed less improvement in knee joint degeneration, higher Lequesne MG score, K-L grade, Pelletier score and Mankin score (P<0.05), higher contents of TNF-α, IL-1β, IL-18, and MMP-13 in knee cartilage tissue and joint fluid (P<0.05), lower mRNA levels and protein relative expressions of A3AR in knee cartilage tissue (P<0.05), higher mRNA levels and protein relative expressions of Wnt3a, β-catenin, NLRP3, Caspase-1 and GSDMD in knee cartilage tissue (P<0.05).

Acupotomy can significantly relieve the symptoms of KOA and delay the course of KOA, and its mechanism may be related to activation of A3AR in knee cartilage tissue to inhibit the transduction of Wnt/β-catenin signaling pathway, thereby interfering with chondrocyte pyroptosis, reducing joint inflammation and further alleviating cartilage matrix degradation.

针刀疗法具有暂缓膝骨关节炎（KOA）进程的作用，但其具体作用机制尚不明确。

探讨针刀疗法对KOA的治疗效果及可能作用机制。

2023年5月—2024年2月将40只SPF级健康新西兰白兔采用SPSS 26.0软件随机分为空白组、模型组、针刀组、针刀联合腺苷A3受体（A3AR）拮抗剂组、针刀联合β-连环蛋白（β-catenin）激活剂组，每组8只。空白组大兔不予任何干预，其余各组大兔采用关节腔灌注木瓜蛋白酶联合膝关节强制屈曲位法构建KOA模型。模型组大兔造模完成后不予任何干预，针刀组大兔予以针刀疗法干预，针刀联合A3AR拮抗剂组大兔予以针刀疗法联合耳缘静脉注射A3AR拮抗剂MRS1220（5 mg/kg）干预，针刀联合β-catenin激活剂组大兔予以针刀疗法联合耳缘静脉注射Wnt/β-catenin信号通路激活剂SKL2001（5 mg/kg）干预，1次/周，持续4周。于干预前、后对各组大兔行行为学观察，于干预结束后对各组大兔膝关节行影像学观察，而后观察各组大兔膝关节软骨大体形态变化，采用苏木素-伊红（HE）染色法对软骨组织行病理学观察，并分别通过Lequesne MG评分、Kellgren-Lawrence（K-L）分级、Pelletier评分及Mankin评分进行量化评估，同时采用酶联免疫吸附试验（ELISA）检测膝关节软骨组织及关节液促炎症细胞因子血清肿瘤坏死因子α（TNF-α）、白介素1β（IL-1β）、白介素18（IL-18）、基质金属蛋白酶-13（MMP-13）含量，采用Real-time PCR法检测膝关节软骨组织A3AR、Wnt3a、β-catenin、Nod样受体蛋白3（NLRP3）、半胱氨酸蛋白酶1（Caspase-1）、消皮素D（GSDMD）mRNA水平，采用Western Blotting法检测膝关节软骨组织A3AR、Wnt3a、β-catenin、NLRP3、Caspase-1、GSDMD蛋白相对表达量。

与空白组比较，模型组膝关节退变明显，Lequesne MG评分、K-L分级、Pelletier评分及Mankin评分升高（P<0.05），膝关节软骨组织及关节液TNF-α、IL-1β、IL-18、MMP-13含量升高（P<0.05），膝关节软骨组织A3AR mRNA水平及蛋白相对表达量降低（P<0.05），膝关节软骨组织Wnt3a、β-catenin、NLRP3、Caspase-1、GSDMD mRNA水平及蛋白相对表达量升高（P<0.05）；与模型组比较，针刀组上述指标皆上升或下降，趋于恢复至正常范围（P<0.05）；与针刀组比较，针刀联合A3AR拮抗剂组、针刀联合β-catenin激活剂组膝关节退变改善程度明显下降，Lequesne MG评分、K-L分级、Pelletier评分及Mankin评分升高（P<0.05），膝关节软骨组织及关节液TNF-α、IL-1β、IL-18、MMP-13含量升高（P<0.05），膝关节软骨组织A3AR mRNA水平及蛋白相对表达量降低（P<0.05），膝关节软骨组织Wnt3a、β-catenin、NLRP3、Caspase-1、GSDMD mRNA水平及蛋白相对表达量升高（P<0.05）。

针刀疗法可明显改善KOA症状，延缓KOA病程，其机制可能与其活化膝关节软骨组织中的A3AR以抑制Wnt/β-catenin信号通路的转导进而干预软骨细胞焦亡、减轻关节炎症反应并进一步缓解软骨基质降解相关。