电针透刺法通过TLR4/MyD88/NF-κB信号通路改善膝关节骨关节炎滑膜炎性反应的机制研究  

Mechanism of electroacupuncture penetration needling for relieving synovial inflammation of knee osteoarthritis through TLR4/MyD88/NF-κB signal pathway

作　　者：周子齐 杨永菊 马贤德[1] 张师侥 关雪峰 ZHOU Zi-qi;YANG Yong-ju;MA Xian-de;ZHANG Shi-yao;GUAN Xue-feng(Graduate School of Liaoning University of Traditional Chinese Medicine,Shenyang 110000;No.2 Department of Orthopedic Rehabilitation,Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110000)

机构地区：[1]辽宁中医药大学研究生学院,沈阳110000 [2]辽宁中医药大学附属医院骨伤康复二科,沈阳110000

出　　处：《针刺研究》2023年第4期353-358,365,共7页Acupuncture Research

基　　金：辽宁省中医药科学院平台建设项目(No.2019JH6/10100002)。

摘　　要：目的:观察电针透刺法对大鼠滑膜Toll样受体4/髓样分化因子88/核转录因子-κB(TLR4/MyD88/NF-κB)信号通路及血清相关炎性因子的影响,探讨电针透刺法对膝关节骨关节炎(KOA)大鼠滑膜炎性反应的影响机制。方法:雄性SD大鼠随机分为假手术组、模型组、电针透刺组、普通电针组,每组16只。采用前交叉韧带离断术结合术后强迫运动制备KOA大鼠模型。电针透刺组于右侧“犊鼻”“内膝眼”予相互透刺5~8 mm,“血海”“梁丘”予相互透刺5~8 mm;普通电针组于右侧“犊鼻”“内膝眼”予向上与皮肤呈30°角斜刺3~5 mm,“血海”“梁丘”予直刺3~5 mm。两组均同样连接电针,疏密波,频率2 Hz/10 Hz,强度0.5~1.5 mA,1次/d,每次20 min,10 d为1个疗程,每疗程间休息2 d,共2个疗程。肌骨超声观察大鼠膝关节积液情况;ELISA法检测大鼠血清IL-1β、IL-6及TNF-α的含量;HE染色法观察大鼠滑膜形态学变化;免疫组织化学法检测大鼠滑膜组织中NF-κB p65的阳性表达;Western blot法检测大鼠滑膜组织中TLR4、MyD88、肿瘤坏死因子受体相关因子-6(TRAF-6)及NF-κB p65蛋白表达水平。结果:与假手术组比较,模型组膝关节积液明显增多,滑膜内衬层细胞分布不规则,细胞排列紊乱,生成大量血管翳,可见大量炎性细胞浸润,血清IL-1β、IL-6、TNF-α含量及滑膜组织中NF-κB p65阳性表达增加(P<0.05),TLR4、MyD88、TRAF-6、NF-κB p65蛋白表达水平升高(P<0.05);与模型组比较,电针透刺组和普通电针组膝关节积液明显减少,可见滑膜内衬层细胞增生、少量的炎性细胞浸润和血管翳生成,血清IL-1β、IL-6、TNF-α含量及滑膜组织NF-κB p65阳性表达减少(P<0.05),TLR4、MyD88、TRAF-6、NF-κB p65蛋白表达水平降低(P<0.05);与电针透刺组比较,普通电针组膝关节积液较多,血清IL-1β、IL-6、TNF-α含量及滑膜组织TLR4、MyD88、NF-κB p65蛋白表达水平均升高(P<0.05)。结论:电针透刺法�Objective To observe the effects of electroacupuncture(EA) penetration needling on Toll-like receptors 4/myeloid differentiation factor 88/nuclear factor-kappa B(TLR4/MyD88/NF-κB) signaling pathway in rat synovium and the serum-related inflammatory factors,so as to explore the mechanism of EA penetration needling on synovial inflammation in rats with knee osteoarthritis(KOA).Methods SD male rats were randomly divided into sham-operation group,model group,EA+penetration needling group,and conventional EA group,with 16 rats in each group.The rats model was prepared by anterior cruciate ligment transection and these rats were forced to exercise for 8 weeks after operation.After successful modeling,in the EA+penetration needling group,the needles were inserted at “Dubi”(ST35) “Neixiyan”(EX-LE4),and at “Xuehai”(SP10) “Liangqiu”(ST34) on the right hind limb,towards each other,5-8 mm in depth,respectively.In the conventional EA group,the needles were inserted at ST35 and EX-LE4 on the right hind limb,obliquely,at 30° angle to the skin,3-5 mm in depth;and were inserted at SP10 and ST34 on the right hind limb perpendicularly,3-5 mm in depth.In these two groups,electric stimulation was operated with dense-disperse wave,2 Hz/10 Hz in frequency and 0.5-1.5 mA in intensity,retained for 20 min in each treatment.The treatment was given once daily,10 days as 1 course of treatment,and 2 courses were required at the interval of 2 days.After the intervention,the knee joint effusion was observed by musculoskeletal ultrasound;the contents of IL-1β,IL-6 and TNF-α in serum were determined by ELISA;the morphological changes in the synovium were observed after H.E.staining;the positive expression of NF-κB p65 in the synovial membrane was detected by immunohistochemical method;the expression levels of TLR4,MyD88,TRAF-6 and NF-κB p65 proteins in the synovial membrane were determined by Western blot.Results Compared with the sham-operation group,in the model group,the knee joint effusion was obviously increased,the syn

关 键 词：膝关节骨关节炎 电针 透刺 Toll样受体4 髓样分化因子88 核转录因子-κB 滑膜 

分 类 号：R245.97[医药卫生—针灸推拿学]