针刀疏筋解结术对类风湿关节炎家兔滑膜炎症的影响及作用机制研究  

Study on the Mechanism and Effects of Needle-knife Tendon-sparing and Knot-dissolving Technique on Synovial Inflammation of Rheumatoid Arthritis Rabbits

作　　者：陈平[1] 王海东[1] 杜小正[2] 井维尧 刘翠 李浩林 陶鹏飞 CHEN Ping;WANG Haidong;DU Xiaozheng;JING Weiyao;LIU Cui;LI Haolin;TAO Pengfei(Rheumatism and Bone Disease Center,Gansu Provincial Hospital of Traditional Chinese Medicine,Lanzhou 730050,China;Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃省中医院风湿骨病中心,甘肃兰州730050 [2]甘肃中医药大学,甘肃兰州730000

出　　处：《中国中医药信息杂志》2024年第7期91-99,共9页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金地区科学基金(82260880);甘肃省自然科学基金(22JR11RA198);甘肃省针刀医学产学研合作平台建设(2022-3-33);王海东甘肃省名中医传承工作室建设项目(2022年)。

摘　　要：目的观察针刀疏筋解结术对类风湿关节炎(RA)模型家兔膝关节滑膜组织NF-κB/Bcl-2通路活性及肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6、Bax、caspase-3表达的影响,探讨其抑制RA滑膜炎症的作用机制。方法24只新西兰白兔随机分为正常组、模型组、药物组和针刀组,每组6只,除正常组外,其余组采用卵蛋白+弗氏完全佐剂膝关节腔注射复制RA模型,分别予相应干预,连续18 d。测定家兔膝关节痛阈、膝关节周径,超声观察关节腔积液、滑膜厚度及内部血流信号,HE染色观察膝关节滑膜组织形态,TUNEL染色观察滑膜组织成纤维样滑膜细胞(FLS)凋亡情况,免疫组化检测滑膜组织TNF-α、IL-1β、IL-6表达,RT-PCR检测滑膜组织核因子(NF)-κBp65、Bcl-2mRNA表达,Westernblot检测滑膜组织NF-κBp65、p-NF-κBp65、Bcl-2、Bax、caspase-3蛋白表达。结果与正常组比较,模型组家兔膝关节痛阈降低、膝关节周径增加,超声评分和滑膜组织病理评分增加;滑膜组织FLS凋亡率降低,TNF-α、IL-1β、IL-6表达升高,NF-κBp65、Bcl-2 mRNA和蛋白及p-NF-κBp65蛋白表达升高,Bax、caspase-3蛋白表达降低,差异均有统计学意义(P<0.05)。与模型组比较,药物组和针刀组家兔膝关节痛阈增加、膝关节周径减小,超声评分和滑膜组织病理评分减少;滑膜组织FLS凋亡率升高,TNF-α、IL-1β、IL-6表达降低,NF-κBp65、Bcl-2m RNA和蛋白及p-NF-κBp65蛋白表达降低,Bax、caspase-3蛋白表达升高,差异均有统计学意义(P<0.05)。结论针刀疏筋解结术可能通过降低滑膜组织NF-κB/Bcl-2通路活性,促进FLS凋亡,减少TNF-α、IL-1β、IL-6产生,抑制RA滑膜炎症,减轻膝关节肿胀、提升痛阈。Objective To observe the effects of needle-knife tendon-sparing and knot-dissolving technique on NF-κB/Bcl-2 pathway activity and the expression of TNF-α,IL-1β,IL-6,Bax and caspase-3 in synovial tissue of knee joints of rabbits with rheumatoid arthritis(RA)model;To explore the mechanism of its inhibitory effect on RA synovial inflammation.Methods Totally 24 New Zealand white rabbits were randomly divided into normal group,model group,drug group and needle-knife group,with 6 rabbits in each group.Except for the normal group,the other groups were injected with ovalbumin and Freund’s complete adjuvant into the knee joint cavity to replicate the RA model,and corresponding interventions were given for 18 consecutive days.The knee pain threshold and knee circumference were measured,ultrasound was used to observe the joint cavity effusion,synovial thickness and internal blood flow signals,the synovial tissue morphology were observed by HE staining,the apoptosis of fibroblast like synoviocytes(FLS)in synovial tissue was observed by TUNEL staining,the expression of TNF-α,IL-1βand IL-6 in synovial tissue were detected by immunohistochemistry,RT-PCR was used to detect NF-κBp65 and Bcl-2 mRNA expression in synovial tissue,Western blot was used to detect the protein expression of NF-κBp65,p-NF-κBp65,Bcl-2,Bax and caspase-3 in synovial tissue.Results Compared with the normal group,the knee pain threshold decreased,knee circumference increased,ultrasonic score and pathological score of synovial tissue increased in the model group;FLS apoptosis rate decreased,the expression of TNF-α,IL-1βand IL-6 in synovial tissue increased,the expressions of NF-κBp65,Bcl-2 mRNA and protein,and p-NF-κBp65 protein increased,while the expression of Bax,caspase-3 protein decreased,with statistical significance(P<0.05).Compared with the model group,the knee pain threshold increased in the drug group and the needle knife group,the circumference of the knee joint decreased,the ultrasonic score and pathological score of synovial tissue w

关 键 词：针刀疏筋解结术 类风湿关节炎 成纤维样滑膜细胞 滑膜炎症 兔 

分 类 号：R245[医药卫生—针灸推拿学]