【目的】 探讨电针透刺法对膝骨关节炎（KOA）大鼠骨代谢的影响及其机制。【方法】 采用关节腔内注射碘乙酸钠法构建 KOA大鼠模型。将SD大鼠随机分为正常组、模型组、塞来昔布组（灌胃塞来昔布）、电针透刺组（针刺犊鼻、内膝眼、血海 和梁丘穴）和3-甲基腺嘌呤（3-MA）（自噬抑制剂）组（腹腔注射3-MA，针刺犊鼻、内膝眼、血海和梁丘穴）。造模后和治疗结 束后分别进行Lequesne MG评分以评价大鼠的行为学变化。治疗结束后，采用苏木精-伊红（HE）染色法观察膝关节软骨组织 病理变化并进行评分，酶联免疫吸附分析（ELISA）检测血清骨钙素（BGP）、Ⅰ 型胶原 C末端肽（CTX-Ⅰ）和抗酒石酸酸性磷 酸酶（TRACP）等骨代谢相关指标，逆转录实时荧光定量聚合酶链式反应（RT-qPCR）法检测膝关节软骨组织中自噬相关蛋白 ULK1、Beclin-1、LC3B、p62水平，Western Blot 法检测膝关节软骨组织中自噬相关蛋白 ULK1、Beclin-1、LC3B、p62蛋白 表达水平和白细胞介素 1β（IL-1β）/雌激素受体相关蛋白 α抗体（ERRα）/SRY相关蛋白 9（SOX9）/Ⅱ型胶原的 α1链（Col2α1）信 号通路相关蛋白表达水平。【结果】 与正常组比较，模型组大鼠关节软骨粗糙不整、结构缺损、排列紊乱，Lequesne MG 评 分，Mankin评分，TRACP、CTX-Ⅰ及p62、IL-1β水平显著上升（P＜0.05），BGP及ULK1、Beclin-1、LC3B、ERRα、SOX9、 Col2α1水平显著降低（P＜0.05）；与模型组比较，塞来昔布组、电针透刺组和3-MA组大鼠软骨表面分层清晰、软骨细胞排 列较为整齐，Lequesne MG 评分，Mankin 评分，TRACP、CTX-Ⅰ及 p62、IL-1β 水平显著降低（P＜0.05），BGP 及 ULK1、 Beclin-1、LC3B、ERRα、SOX9、Col2α1水平显著上升（P＜0.05）；与塞来昔布组比较，电针透刺组和3-MA组大鼠Lequesne MG 评分，Mankin 评分，TRACP、CTX-Ⅰ及 p62、IL-1β 水平显著降低（P＜0.05），BGP 及 ULK1、Beclin-1、LC3B、ERRα、 SOX9、Col2α1水平显著上升（P＜0.05）；与3-MA组，电针透刺组大鼠Lequesne MG评分，Mankin评分，TRACP、CTX-Ⅰ及 p62、IL-1β水平显著降低（P＜0.05），BGP及ULK1、Beclin-1、LC3B、ERRα、SOX9、Col2α1水平显著上升（P＜0.05）。【结论】 电针透刺法能够改善 KOA 大鼠骨代谢，促进自噬，减轻膝关节组织病理损伤，其作用机制可能与抑制 IL-1β/ERRα/SOX9/ Col2α1信号通路有关。

Objective To investigate the effect of electroacupuncture with penetrating needle technique on bone metabolism in rats with knee osteoarthritis （KOA） and its underlying mechanism. Methods A KOA rat model was established by intra-articular injection of sodium monoiodoacetate. SD rats were randomly divided into a normal group，a model group，a celecoxib group （oral gavage of celecoxib），an electroacupuncture penetrating needle group （acupuncture at Dubi [ST35]， Neixiyan [EX-LE5]， Xuehai [SP10]， and Liangqiu [ST34]）， and a 3- methyladenine （3-MA，an autophagy inhibitor） group （intraperitoneal injection of 3-MA plus acupuncture at the same acupoints）. The Lequesne MG index was used to assess behavioral changes in rats after modeling and after treatment. After treatment， pathological changes in knee joint cartilage were observed and scored using hematoxylin-eosin （HE） staining. Serum levels of bone metabolism-related markers， including osteocalcin （BGP），C-terminal telopeptide of type I collagen （CTX-I），and tartrate-resistant acid phosphatase （TRACP）， were detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA levels of ULK1，Beclin-1，LC3B， and p62 in knee cartilage were measured by reverse transcription-quantitative polymerase chain reaction （RT-qPCR）. Protein expression levels of autophagy-related proteins （ULK1，Beclin-1，LC3B，p62） and proteins related to the interleukin-1β （IL-1β）/estrogen receptor-related receptor α （ERRα）/SRY-box transcription factor 9 （SOX9）/type II collagen alpha 1 chain （Col2α1） signaling pathway in knee cartilage were detected by Western Blot. Results Compared with the normal group，the model group exhibited rough，defective，and disorganized articular cartilage structure. The Lequesne MG score，Mankin score，levels of TRACP，CTX-I，p62，and IL-1β were significantly increased （P＜0.05），while levels of BGP，ULK1，Beclin-1，LC3B，ERRα，SOX9，and Col2α1 were significantly decreased （P＜0.05）. Compared with the model group， the celecoxib group， electroacupuncture penetrating needle group， and 3-MA group showed clear cartilage surface stratification and more orderly chondrocyte arrangement. The Lequesne MG score，Mankin score，levels of TRACP，CTX-I，p62， and IL-1β were significantly decreased （P＜0.05），while levels of BGP，ULK1，Beclin-1，LC3B，ERRα， SOX9， and Col2α1 were significantly increased （P＜0.05）. Compared with the celecoxib group， the electroacupuncture penetrating needle group and 3-MA group showed significantly low Lequesne MG score， Mankin score， and levels of TRACP， CTX-I， p62， and IL-1β（P＜0.05）， and significantly high levels of BGP，ULK1，Beclin-1，LC3B，ERRα，SOX9，and Col2α1（P＜0.05）. Compared with the 3-MA group，the electroacupuncture penetrating needle group showed significantly low Lequesne MG score， Mankin score， and levels of TRACP，CTX-I，p62，and IL-1β（P＜0.05），and significantly high levels of BGP，ULK1，Beclin-1， LC3B， ERRα， SOX9， and Col2α1（P＜0.05）. Conclusion Electroacupuncture with penetrating needle technique can improve bone metabolism， promote autophagy， and alleviate pathological damage in knee joint tissue in KOA rats. Its mechanism of action may be related to the inhibition of the IL-1β/ERRα/SOX9/Col2α1 signaling pathway.

R245

广东省医学科学技术研究基金项目（编号：A202208201）