文献类型：期刊文献

中文题名：热补针法对类风湿关节炎寒证家兔膝关节滑膜Keap1-Nrf2/ARE/HO-1信号通路的影响

英文题名：Heat-tonifying acupuncture relieves pain and synovial inflammatory injury by regulating Keap1-Nrf2/ARE/HO-1signaling pathway in rabbits with cold syndrome type rheumatoid arthritis

作者：张枫帆[1];郑欣[2];余俊玲[3];年芳红[1];袁博[4];金芳梅[1];刘雪君[1];田雪梅[1];王海东[1];杜小正[3]

第一作者：张枫帆

机构：[1]甘肃省中医院风湿骨病中心,兰州730000;[2]河西学院医学院,甘肃张掖734000;[3]甘肃中医药大学研究生院,兰州730000;[4]甘肃中医药大学附属医院针灸中心,兰州730000

第一机构：甘肃省中医院风湿骨病中心,兰州730000

年份：2023

卷号：48

期号：7

起止页码：650

中文期刊名：针刺研究

外文期刊名：Acupuncture Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；PubMed;

基金：国家自然科学基金地区科学基金项目(No.82060891)

语种：中文

中文关键词：类风湿关节炎;热补针法;氧化应激;Kelch样环氧氯丙烷相关蛋白-1-核因子E2相关因子2/抗氧化元件/血红素加氧酶信号转导通路

外文关键词：Rheumatoid arthritis;Heat-tonifying needling;Oxidative stress;Keap1-Nrf2/ARE/HO-1signaling pathway

摘要：目的:观察热补针法对类风湿关节炎(RA)寒证家兔膝关节滑膜组织Kelch样环氧氯丙烷相关蛋白-1(Keap1)-核因子E2相关因子2(Nrf2)/抗氧化元件(ARE)/血红素加氧酶(HO-1)信号转导通路的影响,从氧化应激角度探讨热补针法治疗RA的机制。方法:新西兰兔随机分为正常组、模型组、平补平泻组、捻转补法组、热补针法组,每组6只。通过卵蛋白诱导联合低温冷冻法复制RA寒证模型。平补平泻组、捻转补法组、热补针法组分别于双侧“足三里”施以平补平泻法、捻转补法及热补针法,每次30 min,1次/d,治疗7 d。治疗后观察各组家兔的一般情况;测定各组家兔膝关节周径、痛阈值;彩色超声诊断仪观察家兔膝关节滑膜变化并进行影像评级;HE染色观察各组家兔膝关节滑膜组织病理形态并评分;荧量定量PCR法检测各组家兔膝关节滑膜组织Keap1、Nrf2、HO-1、谷胱甘肽氧化酶(GSH-PX)1 mRNA表达水平;Western blot法检测各组家兔膝关节滑膜组织HO-1蛋白表达量。结果:正常组家兔摄食、饮水正常,活动灵敏,精神状态良好;模型组家兔肢体蜷缩,精神萎靡,脱毛严重,活动量小,饮食、饮水量少,皮肤青紫,双后腿蜷曲跛行,触之躲避行为明显。针刺后,各针刺组家兔较模型组一般情况均好转、活动量增大。与正常组比较,模型组家兔膝关节周径明显增大(P<0.05);与模型组比较,各针刺组膝关节周径缩小(P<0.05)。与正常组比较,模型组家兔的膝关节病理积分,滑膜组织Nrf2 mRNA、HO-1 mRNA及蛋白表达升高(P<0.05),滑膜组织Keap1、GSH-PX1 mRNA表达量和痛阈降低(P<0.05)。与模型组比较,各针刺组痛阈,滑膜组织Nrf2、GSH-PX1、HO-1 mRNA及HO-1蛋白表达升高(P<0.05),膝关节周径、病理积分及滑膜组织Keap1 mRNA表达量降低(P<0.05)。热补针法组痛阈,Nrf2、GSH-PX1、HO-1 mRNA及HO-1蛋白表达高于平补平泻组和捻转补法组(P<0.05),膝关节病理积分及滑膜Keap1 mRNA表达量低于平补平泻组和捻转补法组(P<0.05)。结论:热补针法可能通过调控Keap1-Nrf2/ARE/HO-1信号转导通路提高机体抗氧化应激能力,是其治疗RA的机制之一。
Objective To observe the effect of heat-tonifying needling on Keap1-Nrf2/ARE/HO-1 signal transduction pathway in knee synovium in rabbits with cold syndrome type rheumatoid arthritis(RA),so as to explore its mechanisms underl-ying improvement of RA.Methods New Zealand rabbits were randomly divided into normal control,RA model,uniform reinforcing-reducing acupuncture,twisting reinforcing acupuncture and heat-tonifying acupuncture groups,with 6 rabbits in each group.The cold syndrome type RA model was established by subcutaneous injection of mixture fluid of ovalbumin and Freund's complete adjuvant at the shoulder-back as well as injection of mixture of ovalbumin and normal saline into knee-joint cavity combined with ice-compress freezing.Acupuncture stimulation(uniform reinforcing-reducing,or twisting reinforcing or heat-tonifying)was applied to bilateral“Zusanli”(ST36)for 1 min with the needle retained for 30 min,once a day for 7 consecutive days.The general conditions of rabbits in each group were recorded,the thermal pain threshold(TPT)and perimeter of knee joints was measured.Conditions of the synovium in the knee cavity,hydrops,blood flow signal,articular surface,and related muscles were observed by using a color Doppler ultrasonic diagnostic apparatus,and the blood flow signals inside the synovium(image scores)were divided into 0(no signals),I(1 or 2 dot-like signal),II(less than half)ad III(more than half).After H.E.staining,the pathological changes(0-3 points)were assessed according to the state of inflammatory cell infiltration,and hyperplasia of synovial matrix and coating cells.The expression levels of Keap1,Nrf2,HO-1 and GSH-PX1 mRNAs in the knee synovium were detected by quantitative real-time PCR,and the expression of knee synovial HO-1 protein was measured by Western blot.Results In comparison with the normal control group,the model group had a significant increase in the perimeter,pathological score,expression of Nrf2,HO-1 mRNAs and HO-1 protein(P<0.05),and an obvious decrease in the TPT,expression levels of Keap1 and GSH-PX1 mRNAs(P<0.05).Relevant to the model group,all the three acupuncture maneuvers reversed modeling-induced increase of perimeter and pathological score(P<0.05),decrease of TPT and expression of GSH-PX1 mRNA(P<0.05),further down-regulated expression of Keap1 mRNA(P<0.05),further up-regulated the expression of Nrf2,HO-1 mRNAs and HO-1 protein(P<0.05).The heat-reinforcing manipulation was significantly superior to uniform reinforcing-reducing and twirling reinforcing manipulations in up-regulating TPT,and expression of Nrf2 mRNA,GSH-PX1 mRNA,HO-1 mRNA and protein(P<0.05),and in down-regulating pathological score and Keap1 mRNA expression(P<0.05).Conclusion Heat-tonifying,uniform reinforcing-reducing and twirling reinforcing needling manipulations may relieve pain and improve pathological state in RA rabbits,which may be associated with their functions in raising the ability of anti-oxidative stress by regulating Keap1-Nrf2/ARE/HO-1 signaling pathway,the therapeutic effect of heat-tonifying needling is superior to that of uniform reinforcing-reducing and twirling reinforcing needling.