针刺改善Ⅱ型胶原蛋白诱导性关节炎大鼠炎性反应的机制研究  

Effect of acupuncture on arthritic pathological injury in rats with type Ⅱ collagen-induced rheumatoid arthritis

作　　者：李秀芳 马学红[2] 周蓉 刘晨 刘淑清 LI Xiu-fang;MA Xue-hong;ZHOU Rong;LIU Chen;LIU Shu-qing(The Second Clinical College of Beijing University of Chinese Medicine,Beijing 100029,China;Department of Acupuncture-moxibustion,Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078)

机构地区：[1]北京中医药大学第二临床医学院,北京100029 [2]北京中医药大学东方医院针灸科,北京100078

出　　处：《针刺研究》2023年第11期1103-1110,共8页Acupuncture Research

基　　金：北京市科技计划项目(No.Z121100005512024);北京中医药大学校级课题纵向科研发展基金项目(No.2022BZYLL0606)。

摘　　要：目的:观察针刺“阳陵泉”“足三里”“悬钟”对Ⅱ型胶原蛋白诱导性关节炎(CIA)大鼠关节炎指数(AI)、关节滑膜病理形态、血清相关免疫炎性因子及关节滑膜组织突变型p53(mt-p53)、核转录因子-κB(NF-κB)p65、滑膜过氧化物酶体增殖物激活受体-γ(PPARγ)蛋白表达的影响,探讨针刺治疗类风湿性关节炎(RA)的可能机制。方法:随机选取6只雄性SD大鼠作为正常组,其余大鼠采用牛源性Ⅱ型胶原蛋白和完全弗氏佐剂建立CIA模型。将造模成功的18只大鼠随机分为模型组、来氟米特组、针刺组,每组6只。来氟米特组予来氟米特1.9 mg/kg灌胃,每日1次;针刺组针刺“阳陵泉”“足三里”“悬钟”,留针30 min,每日1次。以上均干预12周。每周进行关节炎指数(AI)评分;ELISA法检测血清中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-17含量;HE染色法观察大鼠踝关节的病理学变化;Western blot法检测膝关节滑膜组织中mt-p53、NF-κB p65、PPARγ蛋白表达水平;荧光定量PCR法检测膝关节滑膜组织中mt-p53、NF-κB p65、PPARγ mRNA表达水平。结果:与正常组比较,模型组大鼠AI评分升高(P<0.01,P<0.05),血清中TNF-α、IL-6、IL-17含量升高(P<0.01),膝关节滑膜组织中mt-p53、NF-κB p65、PPARγ蛋白及mRNA表达水平均升高(P<0.01,P<0.05)。与模型组比较,针刺组和来氟米特组大鼠AI评分降低(P<0.01,P<0.05),血清TNF-α、IL-6及IL-17含量降低(P<0.01),关节滑膜组织中NF-κB p65蛋白及mRNA表达水平均降低(P<0.01),PPARγ蛋白表达水平升高(P<0.05);来氟米特组mt-p53蛋白和mRNA表达水平降低(P<0.01),PPARγ mRNA表达水平升高(P<0.01)。与来氟米特组比较,针刺组大鼠滑膜组织中mt-p53、NF-κB p65蛋白及mRNA表达水平升高(P<0.01),PPARγ mRNA表达水平下降(P<0.01)。HE染色结果显示,模型组大鼠踝关节软骨增生,关节腔缩小,周围软组织见轻度纤维增生和炎性细胞浸润;针刺组和来氟米�Objective To observe the effect of acupuncture stimulation of“Yanglingquan”(GB34),“Zusanli”(ST36)and“Xuanzhong”(GB39)on arthritis index(AI),joint synovial membrane pathology,serum-related immuno⁃inflammatory factors,and expressions of tumor suppressor gene mt-p53,nuclear factor kappa B(NF-κB)and peroxi⁃some proliferator activated receptor gamma(PPARγ)in knee joint synovial tissue of rats with typeⅡcollagen-induced arthritis(CIA),so as to explore its possible mechanisms underlying improvement of rheumatoid arthritis(RA).Methods Male SD rats were used in the present study.The CIA model was established by subcutaneous injection of collagen emulsion(200μL/rat)in the tail root region on the first day and repeat(100μL/rat)once on the 9th day.Eigh⁃teen successful CIA rats were randomized into model,medication and acupuncture groups,with 6 rats in each group.Other 6 normal rats were used as the normal control group.For rats of the medication group,leflunomide(1.9 mg/kg)was administrated by gavage,once a day,and for rats of the acupuncture group,manual acupuncture stimulation was applied to bilateral GB34,ST36,GB39 for 30 min,once a day,for 12 weeks.The arthritis index(AI)score(0—4 points)was evaluated once every week.The contents of IL-6,IL-17 and TNF-αin the serum were determined by ELISA.Histopathological changes of the ankle joint were observed by H.E.staining.The protein and mRNA expression levels of mt-p53,NF-κB p65,and PPARγin the knee joint synovial tissue were determined by Western blot and quanti⁃tative real time PCR,separately.Results Compared with the normal control group,the AI scores at different timepoints after modeling,contents of serum TNF-α,IL-6 and IL-17,expression levels of mt-p53,NF-κB p65,PPARγpro⁃teins and mRNAs were significantly increased in the model group(P<0.01,P<0.05).In comparison with the model group,the AI scores at the 10th week in the medication group and at the 3rd,9th and 10th week in the acupuncture group,contents of serum TNF-α,IL-6 and IL-17,and

关 键 词：类风湿性关节炎 针刺 炎性因子 过氧化物酶体增殖物激活受体-Γ 核转录因子-ΚB 突变型P53 

分 类 号：R246.2[医药卫生—针灸推拿学]