摘要
目的:观察艾灸对类风湿性关节炎(RA)大鼠踝关节滑膜组织Toll样受体4/核因子-κB (TLR 4/NF-κB)信号通路的影响,分析艾灸治疗RA的生物学机制。方法:SD大鼠随机分为正常组、模型组、艾灸组、激动组、拮抗组,每组10只。风、寒、湿环境+弗氏完全佐剂复制RA模型。造模3d后对艾灸组、激动组和拮抗组大鼠双侧"肾俞""足三里"穴各施灸20min,激动组和拮抗组分别在艾灸前30min经尾静脉注射TLR 4激动剂或拮抗剂。采用Western blot法测定各组大鼠踝关节滑膜组织NF-κB抑制因子α(IκBα)、IκB激酶复合物β(IκKβ)、髓样分化因子88(MyD 88)、TLR 4、NF-κB p 65蛋白的表达水平。结果:与正常组比较,造模后大鼠右后踝关节肿胀显著(P<0.01),踝关节滑膜组织IκBα、IκKβ、TLR 4、MyD 88、NF-κB p 65的表达水平均显著升高(P<0.01)。与模型组比较,各治疗干预组大鼠右后踝关节肿胀度和踝关节滑膜组织IκKα、IκBβ、TLR 4、MyD 88、NF-κB p 65的表达水平均明显下降(P<0.01,P<0.05)。与艾灸组比较,激动组右后踝关节肿胀度及踝关节滑膜组织IκBα、IκKβ、TLR 4、MyD 88、NF-κB p 65表达水平均升高(P<0.05,P<0.01);拮抗组踝关节滑膜组织IκBα、TLR 4、MyD 88、NF-κB p 65的表达水平显著降低(P<0.01)。结论:艾灸能够显著减轻RA大鼠关节肿胀程度,RA的发病及艾灸对其治疗作用与TLR 4/NF-κB信号通路密切相关,艾灸可以有效抑制RA大鼠TLR 4/NF-κB活化的信号途径,下调相关炎性因子的表达。
Objective To observe the effect of moxibustion on Toll-like receptor 4/nuclear factor-κB（TLR 4/NF-κB）signaling in the synovial tissue of the ankle joint in rats with rheumatoid arthritis（RA）,so as to analyze its biological mechanism underlying improvement of RA.Methods Fifty male SD rats were randomly divided into normal control,model,moxibustion,moxibustion＋ TLR 4 agonist,and moxibustion＋ TLR 4 antagonist groups（n=10 rats in each）.The RA model was established by subcutaneous injection of Freund＇s complete adjuvant（FCA,0.1 mL/rat）at the right hind-paw and by being raised in a wind（air fan blowing）,cold（about 10℃）and wet（purling）environment for 20 days.After 3 days of modeling,mild moxibustion was applied to bilateral＂Shenshu＂（BL 23）and＂Zusanli＂（ST 36）for 20 minutes,once daily for successive 10 days.The TLR 4 agonist（lipopolysaccharide）or TLR 4 antagonist（TAK-242）（1 mg/mL）was separately administered via the tail vein 30 min before performing moxibustion every time in the agonist group and the antagonist group.The expression of NF-κB inhibitory factorɑ（IκBɑ）,IκB kinase complexβ（IκKβ）,myeloid differentiation factor 88（MyD 88）,TLR 4,and NF-κB p 65 proteins in the synovial tissue of the ankle joint was detected by using Western blot.Results Following modeling,the girth of the swollen ankle joint was obviously bigger（P〈0.01）,and the expression levels of IκBɑ,IκKβ,TLR 4,MyD 88,and NF-κB p 65 proteins in the synovial tissue were considerably increased in the model group relevant to the normal control group（P〈0.01）.After moxibustion intervention,the girth of the swollen ankle joint and the expression levels of IκBɑ,IκKβ,TLR 4,MyD 88,and NF-κB p 65 proteins were significantly down-regulated in the moxibustion,moxibustion＋TLR 4 agonist,moxibustion＋TLR 4 antagonist groups compared with the model group（P〈0.01,P〈0.05）.Comparison among the 3 moxibustion groups showed that the lowered levels of ankle-joint girth,and IκBɑ,IκKβ,TLR 4,MyD 88,and NF-κB p 65 expression were significantly smaller in the moxibustion＋TLR 4 agonist group than in the simple moxibustion and moxibustion＋TLR 4 antagonist groups（P〈0.05,P〈0.01）.No significant differences were found between the moxibustion and moxibustion＋TLR 4 antagonist groups in the decreased ankle joint girth and IκBɑ,IκKβ,TLR 4,MyD 88,and NF-κB p 65 expression levels（P〈0.05）,suggesting that activation of TLR 4 reduced the anti-inflammatory effect of moxibustion intervention.Conclusion Moxibustion can reduce the ankle joint swelling in RA rats,which may be closely associated with its effect in down-regulating the expression of IκBɑ,IκKβ,TLR 4,MyD 88,and NF-κB p 65 proteins and in inhibiting TLR 4/NF-κB signaling in the synovial tissue of the ankle joint.
作者
张传英
胡玲
蔡荣林
彭传玉
袁娟
ZHANG Chuan-ying;HU Ling;CAI Rong-lin;PENG Chuan-yu;YUAN Juan(School of Nursing;Institute of Acupuncture-moxibustion and Meridian,Anhui University of Chinese Medicine,Hefei 230012,China)
出处
《针刺研究》
CAS
CSCD
北大核心
2018年第11期687-691,共5页
Acupuncture Research
基金
安徽省2016年度自然科学基金项目(No.1608085QH 219)
国家重点基础研究发展计划("973"计划
No.2015CB 554504)
作者简介
张传英，医学硕士，副教授，研究方向：针灸抗炎免疫机制研究。E—mail：1047042901@qq．com
