文献类型：期刊文献

中文题名：基于OPG/RANKL/NF-κB信号通路探讨针刀疏筋解结术抑制胶原诱导性关节炎大鼠骨破坏的作用机制

英文题名：Study on the underlying mechanism of acupotomy in inhibiting bone destruction in collageninduced arthritis rats based on OPG/RANKL/NF-κB signaling pathway

作者：张智敏[1];王海东[2];李浩林[1];金芳梅[2];阚丽丽[2];陈平[2];杨娟娟[1];杨玉萍[1];程伟刚[1];柏倩[1]

第一作者：张智敏

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省中医院,兰州730050

第一机构：甘肃中医药大学

年份：2026

卷号：51

期号：1

起止页码：68

中文期刊名：针刺研究

外文期刊名：Acupuncture Research

收录：;北大核心:【北大核心2023】；

基金：国家自然科学基金项目(No.82460945);兰州市科技计划项目(No.2022-3-33);甘肃省卫生健康委员会王海东甘肃省名中医传承工作室建设项目(No.甘卫中医函[2022]50号);甘肃省中医药科研项目(No.GZKG-2024-45、GZKG-2024-4);泾川县科技计划项目(No.JC-2024-13)。

语种：中文

中文关键词：针刀疏筋解结术;类风湿关节炎;OPG/RANKL/NF-κB信号通路;骨破坏

外文关键词：Acupotomy tendon-sparing and knot-dissolving technique;Rheumatoid arthritis;OPG/RANKL/NFκB signaling pathway;Bone destruction

摘要：目的:探讨针刀疏筋解结术抑制胶原诱导性关节炎(CIA)大鼠骨破坏的作用机制。方法:24只SPF级SD雌性大鼠,随机选取8只为空白组,剩余大鼠采用牛Ⅱ型胶原联合弗氏不完全佐剂诱导CIA模型,成模后随机分为模型组和针刀组,每组8只。针刀组予通蔽启机术和通阳启枢术交替治疗,1次/3 d,共治疗9次。干预结束后,各组大鼠行关节炎指数(AI)评分;Micro-CT观察大鼠踝关节骨破坏情况并分析骨密度(BMD)、骨表面体积比(BS/BV)、骨体积分数(BV/TV)、骨小梁数量(Tb.N)、骨小梁厚度(Tb.Th)和骨小梁分离度(Tb.Sp);抗酒石酸酸性磷酸酶(TRAP)染色法观察大鼠踝关节组织中破骨细胞(OC)数量变化;HE染色法观察大鼠踝关节病理变化;ELISA法检测大鼠血清肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-6、IL-1β含量;实时荧光定量PCR法和Western blot法分别检测大鼠踝关节组织中骨保护素(OPG)、核转录因子-κB受体活化因子配体(RANKL)、核转录因子-κB(NF-κB)p65、核转录因子-κB抑制蛋白激酶β(IKKβ)mRNA和蛋白表达。结果:与空白组比较,模型组AI评分显著升高(P<0.01);Micro-CT显示踝关节表面粗糙,间隙增大,骨量明显减少,骨侵蚀严重,BMD、BV/TV、Tb.Th和Tb.N均显著降低(P<0.01),BS/BV和Tb.Sp均显著升高(P<0.01);踝关节组织中OC数量显著增加(P<0.01);HE染色显示踝关节组织滑膜细胞增生明显,大量炎性细胞浸润;血清TNF-α、IL-6、IL-1β含量显著升高(P<0.01);踝关节组织中OPG蛋白和mRNA相对表达量显著下降(P<0.01),RANKL、NF-κB p65、IKKβ蛋白和mRNA相对表达量显著升高(P<0.01)。与模型组比较,针刀组干预14、21、28 d后AI评分显著降低(P<0.01);Micro-CT显示踝关节表面密度改善,骨量增加,骨侵蚀减轻;BMD、BV/TV、Tb.Th和Tb.N均显著升高(P<0.01),BS/BV和Tb.Sp均显著降低(P<0.01);踝关节组织中OC数量显著减少(P<0.01);HE染色显示踝关节组织滑膜细胞增生不明显,少量炎性细胞浸润;血清TNF-α、IL-6、IL-1β含量显著降低(P<0.01);踝关节组织中OPG蛋白和mRNA相对表达量升高(P<0.01,P<0.05),RANKL、NF-κB p65、IKKβ蛋白和mRNA相对表达量显著降低(P<0.01)。结论:针刀疏筋解结术能够减轻CIA大鼠炎性反应,抑制骨破坏,可能与调控OPG/RANKL/NF-κB信号通路进一步抑制OC分化有关。
Objective To explore the mechanism of acupotomy in inhibiting bone destruction in rats with collagen-induced arthritis(CIA).Methods Twenty-four SD female rats were randomly allocated to blank control,model and acupotomy groups,with 8 rats in each group.The CIA model was established by subcutaneous injection of bovine type Ⅱ collagen+Freund’s incomplete adjuvant at the base of the tail(on day 0 and 7).The rats of the acupotomy group received alternative relaxing treatment of Tongbi Qiji technique(sternum-xiphoid)and Tongyang Qishu technique(the region around Dazhui[GV14])with needle knife,once every 3 d,for a total of 9 sessions.The rats of the blank control and model groups received grabbing and fixing as those of the acupotomy group.After the intervention,the arthritis index(AI)score(0—4 points for each limb,16 points in total)was evaluated.Micro-CT was used to observe the bone destruction of the ankle joint and to analyze the related bone parameters including bone mineral density(BMD),bone surface/bone volume(BS/BV),bone volume/total volume(BV/TV),bone trabecular number(Tb.N),trabecular thickness(Tb.Th)and trabecular separation/spacing(Tb.Sp).The tartrate-resistant acid phosphatase(TRAP)staining was used to observe changes of the number of osteoclast(OC)in the ankle tissue,and the HE staining was used to observe histopathological changes of the ankle joint.The contents of serum tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-1βwere detected by ELISA.The mRNA and protein expression levels of osteoprotegin(OPG),receptor activator of nuclear factor-κB ligand(RANKL),nuclear factor-κB(NF-κB)p65,and nuclear factor-κB inhibitory protein kinaseβ(IKKβ)in the ankle tissue were detected by real-time fluorescent quantitative PCR and Western blot,respectively.Results Compared with the blank control group,the AI score,BS/BV,Tb.Sp,serum contents of TNF-α,IL-6 and IL-1β,and the expression levels of RANKL,NF-κB p65 and IKKβmRNA and protein were significantly increased(P<0.01),while BMD,BV/TV,Tb.Th and Tb.N,and the expression levels of OPG mRNA and protein were all significantly decreased in the model group(P<0.01).After acupotomy intervention,both the increase of the AI score from day 14 to 28 after intervention,BS/BV,Tb.Sp,serum contents of TNF-α,IL-6 and IL-1β,and the expression levels of RANKL,NF-κB p65 and IKKβmRNA and protein,and the decrease of BMD,BV/TV,Tb.Th and Tb.N,and the expression levels of OPG mRNA and protein were reversed in the acupotomy group(P<0.01,P<0.05).Micro-CT showed rough surface of the ankle joint,enlarged gap,significantly reduced bone mass and serious bone erosion in the model group,and improved surface density of the ankle joint,increased bone mass and decreased bone erosion in the acupotomy group.HE staining displayed an obvious proliferation of synovial cells in the ankle joint tissue,and infiltration of a large number of inflammatory cells in the model group;but no apparent proliferation of the synovial cells in the ankle joint tissue,and only a small amount of inflammatory cells in the acupotomy group.Conclusion Acupotomy can reduce inflammation,inhibit bone destruction in RA rats,which may be related to its function in regulating the OPG/RANKL/NF-κB signaling pathway,further inhibiting OC differentiation.