针刺研究

2015, v.40(03) 199-204

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艾灸对类风湿性关节炎大鼠关节滑膜组织Toll样受体4-骨髓样分化因子88-核转录因子-κB信号通路的影响
Influence of Moxibustion on TLR 4-MyD 88-NF-κB Signal Transduction Pathway of Synovial Tissue in Rheumatoid Arthritis Rats

袁娟;胡玲;宋小鸽;唐照亮;张传英;余爽;尤元梅;
YUAN Juan;HU Ling;SONG Xiao-ge;TANG Zhao-liang;ZHANG Chuan-ying;YU Shuang;YOU Yuan-mei;Anhui University of Traditional Chinese Medicine;

摘要(Abstract):

目的:观察艾灸对类风湿性关节炎(RA)大鼠关节滑膜组织Toll样受体4-骨髓样分化因子88-核转录因子-κB(TLR 4-MyD 88-NF-κB)信号通路的影响,进一步探讨艾灸治疗RA的作用机制。方法 :40只SD大鼠随机分为正常组、模型组、艾灸组和药物组,每组10只。采用风、寒、湿环境因素结合弗氏完全佐剂的方法复制RA模型。艾灸组以艾条悬灸大鼠"足三里""肾俞"穴,每天1次,连续治疗15d。药物组以雷公藤溶液灌胃(8.75mL/kg),每天1次,共15d。观察关节滑膜组织病理形态学变化,放射免疫法检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素1(IL-1)含量,实时荧光定量PCR方法检测滑膜组织TLR 4、MyD 88、肿瘤坏死因子受体相关因子6(TRAF-6)mRNA的表达,免疫组化方法检测NF-κB p 65的表达。结果:与正常组比较,模型组滑膜组织有明显的病理改变,大量单核细胞及淋巴细胞等炎性细胞浸润,滑膜表面不整齐,滑膜增生变厚;与模型组比较,艾灸组和药物组均能明显缓解炎性细胞对滑膜的浸润,改善滑膜细胞增生反应。与正常组比较,模型组血清TNF-α、IL-1含量升高(P<0.01),滑膜TLR 4mRNA、MyD 88mRNA、TRAF-6mRNA及NF-κB p 65表达均升高(P<0.01);与模型组比较,艾灸组和药物组血清中TNF-α、IL-1含量降低(P<0.01),滑膜TLR 4mRNA、MyD 88mRNA、TRAF-6mRNA及NF-κB p 65表达均降低(P<0.01,P<0.05)。结论:艾灸具有调节细胞因子的作用,其机制可能与艾灸抑制滑膜组织TLR 4-MyD 88-NF-κB信号通路的异常激活有关。
Objective To observe the effect of moxibustion intervention on the expression of toll-like receptor-4(TLR 4),myeloid differentiation factor 88(MyD 88),and nuclear factor kappa B p 65(NF-κB p 65)genes of knee-joint synovial cells in rheumatoid arthritis(RA)rats,so as to explore its molecular mechanism underlying improving RA.Methods Forty SD rats were randomly divided into normal control,RA model,moxibustion and medication groups(n=10).The RA model was duplicated by raising the rats in a windy,cold and wet environment,followed by injecting Freund's complete adjuvant(0.15mL)into the rat's foot.Moxibustion was applied to"Shenshu"(BL 23)and"Zusanli"(ST 36)for 20 min,once daily for 15 days.Rats of the medication group were treated by intragastric administration of tripterygium wilfordii(8.75mL/kg),once daily for 15 days.Pathological changes of the synovial tissues were detected by H.E.stain,the contents of serum TNF-αand IL-1detected by radioimmunoassay,and the expression levels of TLR 4mRNA,MyD 88 mRNA and TRAF-6mRNA of synovial tissue were analyzed by real time-PCR,and the NF-κB p 65 immunoactivity was assayed by immunohistochemistry.Results In comparison with the normal control group,the synovial tissue of the knee-joint was impaired remarkably and infiltrated by numerous inflammatory cells,and the synovial surface got thickening because of hyperplasia in the model group.Following moxibustion and medication,these situations such as synovial infiltration of inflammatory,synovial cell proliferation were alleviated.The serum TNF-αand IL-1contents and synovial TLR 4mRNA,MyD 88 mRNA,TRAF-6mRNA and NF-κB p 65 expression levels were remarkably higher in the model group than in the control group(P<0.01).After moxibustion and medication,the contents of serum TNF-αand IL-1,the expression levels of TLR 4mRNA,MyD 88 mRNA,TRAF-6mRNA and NF-κB p 65 were significantly down-regulated(P<0.01,P<0.05).Conclusion Moxibustion intervention can improve pathological changes of the knee-joint synovial membrane tissue in RA rats,which may be related with its effect in inhibiting abnormal activation of TLR 4-MyD 88-NF-κB pathway in synovial tissue.

关键词(KeyWords): 艾灸;类风湿性关节炎;滑膜组织;TLR 4-MyD 88-NF-κB通路
Moxibustion;Rheumatoid arthritis;Synovial tissue;TLR 4-MyD 88-NF-κBpathway

Abstract:

Keywords:

基金项目(Foundation): 国家重点基础研究发展计划“973计划”(No.2009CB 522905);; 安徽中医药大学校级自然科学基金(No.2015zr 013)

作者(Authors): 袁娟;胡玲;宋小鸽;唐照亮;张传英;余爽;尤元梅;
YUAN Juan;HU Ling;SONG Xiao-ge;TANG Zhao-liang;ZHANG Chuan-ying;YU Shuang;YOU Yuan-mei;Anhui University of Traditional Chinese Medicine;

DOI: 10.13702/j.1000-0607.2015.03.005

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