摘要
目的:观察电针对类风湿关节炎(RA)大鼠踝关节滑膜组织中自噬体、unc-51样自噬激活激酶1(ULK1)、Beclin1和微管相关蛋白轻链3(LC3)表达及滑膜细胞超微结构的影响,探讨电针通过细胞自噬途径发挥调节细胞增殖作用的机制。方法:将SD大鼠按照随机数字表法分为正常组、模型组、甲氨蝶呤组、电针组,每组6只。采用弗氏完全佐剂造模法制备RA大鼠模型。甲氨蝶呤组予甲氨蝶呤灌胃(0.35 mg/kg),每周2次,治疗4周。电针组予电针"足三里""关元",留针20 min,每日1次,每周休息1 d,治疗4周。足趾容积测量仪测量大鼠左后肢足趾容积,HE染色法观察大鼠踝关节滑膜组织形态,透射电镜观察大鼠踝关节滑膜细胞自噬小体情况,ELISA检测大鼠血清中白细胞介素(IL)-1、肿瘤坏死因子(TNF)-α的含量,Western blot法检测大鼠踝关节滑膜组织ULK1、Beclin1、LC3蛋白的表达。结果:与正常组比较,模型组大鼠左侧足趾容积明显增加(P<0.01),血清中IL-1、TNF-α含量显著升高(P<0.01),踝关节滑膜组织ULK1、Beclin1、LC3蛋白表达显著升高(P<0.01,P<0.05),滑膜增生明显,滑膜细胞可见部分散在分布的双层膜自噬体。与模型组比较,甲氨蝶呤组、电针组左侧足趾容积显著缩小(P<0.01),血清中IL-1、TNF-α含量显著降低(P<0.01),踝关节滑膜组织内ULK1、LC3蛋白表达量降低(P<0.05,P<0.01),滑膜增生改善,滑膜细胞未见明显自噬小体。与甲氨蝶呤组比较,电针组ULK1蛋白表达量显著下降(P<0.01)。结论:电针能改善RA大鼠关节肿胀和踝关节滑膜细胞损伤,其作用机制可能与调节ULK1、LC3、Beclin1等表达,抑制滑膜细胞自噬,调节细胞增殖有关。
Objective To observe the effects of electroacupuncture(EA)on autophagosomes,unc-51-like autophage activating kinase 1(ULK1),Beclin1,and microtubule-associated protein light chain 3(LC3)expression,and synoviocyte ultrastructure in the synovial tissues of rheumatoid arthritis(RA)rats,and to explore the mechanism of EA in regulating the proliferation of synoviocytes via the autophagy pathway.Methods The SD rats were randomly divided into a normal group,a model group,a methotrexate(MTX)group,and an EA group,with 6 rats in each group.Following RA modeling with Freund’s complete adjuvant,rats in the MTX group were treated with intragastric administration of 0.35 mg/kg MTX,twice a week,for 4 weeks,while those in the EA group received 20-min EA stimulation at“Zusanli”(ST36)and“Guanyuan”(CV4),once per day,for 4 weeks,with an interval of one day between weeks.The rat left hind toe volume was measured using the toe volume measuring instrument.HE staining was conducted for detecting the morphology of rat synovial tissues,followed by the observation of autophagosomes under the transmission electron microscope.The levels of serum interleukin(IL)-1 and tumor necrosis factor(TNF)-αwere assayed by ELISA,and the protein expression levels of ULK1,Beclin1,and LC3 were detected by Western blot.Results Compared with the normal group,the left hind toe volume of the model group increased significantly(P<0.01),while serum IL-1 and TNF-α(P<0.01),synovial ULK1,Beclin1,and LC3 protein expression(P<0.01,P<0.05)up-regulated.HE stain and electron microscope showed obvious synovial hyperplasia,and doublemembrane autophagosomes scattering in the synoviocytes.The comparison with the model group showed that MTX and EA remarkably decreased the left hind toe volume(P<0.01),serum IL-1 and TNF-α(P<0.01),down-regulated the protein expression levels of ULK1 and LC3 in synovial tissues(P<0.05,P<0.01),and inhibited the synovial hyperplasia,with no obvious autophagosomes observed in the synoviocytes.The protein expression of ULK1 in the EA gro
作者
刘梨
周巍
黎铭玉
周兰
王文怡
龚志贤
艾坤
LIU Li;ZHOU Wei;LI Ming-yuz;ZHOU Lan;WANG Wen-yi;GONG Zhi-xian;AI Kun(The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;College of Acupuncture-moxibustion and Tuina,Hunan University of Chinese Medicine,Changsha 410208;College of Nursing,Hunan University of Chinese Medicine,Changsha 410208)
出处
《针刺研究》
CAS
CSCD
北大核心
2021年第12期1023-1028,共6页
Acupuncture Research
基金
国家自然科学基金项目(No.81804204,82074565)
湖南省自然科学基金项目(No.2020JJ5433)。
关键词
电针
类风湿性关节炎
滑膜细胞
自噬
自噬相关分子
Electroacupuncture
Rheumatoid arthritis
Synoviocytes
Autophagy
Autophagy-related molecules
