艾灸对佐剂性关节炎大鼠足趾滑膜组织自噬和凋亡的影响  

Effect of moxibustion on the indicators of autophagy and apoptosis in synovium of rats with adjuvant arthritis

作　　者：刘梨[1] 张亮[2] 艾坤[2] 祁芳[2] 李鑫[3] 王文怡 徐美丽 龚志贤[1] LIU Li;ZHANG Liang;AI Kun;QI Fang;LI Xin;WANG Wen-yi;XU Mei-li;GONG Zhi-xian(The First Affiliated Hospital/The First Clinical College of Hunan University of Chinese Medicine,Changsha 410007,China;College of Acupuncture-Moxibustion and Tuina,Changsha 410208;Hunan Provincial Key Laboratory of Diagnostics of Chinese Medicine,Changsha 410208;College of Nursing,Hunan University of Chinese Medicine,Changsha 410208)

机构地区：[1]湖南中医药大学第一附属医院/第一中医临床学院,长沙410007 [2]湖南中医药大学针灸推拿学院,长沙410208 [3]湖南中医药大学中医诊断学湖南省重点实验室,长沙410208 [4]湖南中医药大学护理学院,长沙410208

出　　处：《针刺研究》2023年第3期253-258,共6页Acupuncture Research

基　　金：国家自然科学基金项目(No.81804204、82074565、82274627);湖南省自然科学基金项目(No.2020JJ5433)。

摘　　要：目的:观察艾灸对佐剂性关节炎(AA)大鼠足趾滑膜组织自噬和凋亡相关指标的影响,探讨艾灸治疗类风湿性关节炎的作用机制。方法:SD大鼠随机分为空白组、模型组、艾灸组、甲氨蝶呤组、雷帕霉素组,每组9只。采用弗氏完全佐剂注射法制备AA大鼠模型。艾灸组予艾灸“足三里”“关元”,每次20 min,每日1次;甲氨蝶呤组予甲氨蝶呤(0.35 mg/kg)灌胃,每周2次;雷帕霉素组予雷帕霉素腹腔注射(1 mg/kg),隔日1次;3组均干预3周。采用足趾容积测量仪检测大鼠左后肢足趾容积,ELISA法检测大鼠血清中白细胞介素(IL)-1、肿瘤坏死因子(TNF)-α含量,透射电镜观察大鼠足趾关节滑膜细胞自噬体,Western blot法检测大鼠足趾滑膜组织中哺乳动物雷帕霉素靶蛋白(mTOR)C1、磷酸化(p)-mTORC1、半胱氨酸天冬氨酸蛋白酶(Caspase)-3、死亡受体(Fas)、死亡受体配体(FasL)蛋白表达量。结果:与空白组比较,模型组大鼠足趾容积增大(P<0.01),血清中IL-1、TNF-α含量显著升高(P<0.001),足趾滑膜组织中p-mTORC1蛋白表达量升高(P<0.01),Caspase-3、Fas、FasL蛋白表达量降低(P<0.05,P<0.01),滑膜细胞内自噬小体数量减少。与模型组比较,艾灸组、甲氨蝶呤组大鼠足趾容积明显减小(P<0.05),血清中IL-1、TNF-α含量显著降低(P<0.01,P<0.05,P<0.001),p-mTORC1蛋白表达量降低(P<0.05),Caspase-3、Fas、FasL蛋白表达量升高(P<0.05),滑膜细胞内可见较多自噬小体;雷帕霉素组Caspase-3蛋白表达量升高(P<0.05),滑膜细胞可见较多自噬小体。结论:艾灸能改善AA大鼠关节肿胀,降低血清IL-1、TNF-α含量,其作用机制可能与调节滑膜组织p-mTORC1、Caspase-3、Fas、FasL蛋白表达量从而促进自噬和凋亡有关。Objective To observe the effect of moxibustion on the indicators of autophagy and apoptosis in the synovium of toes of rats with adjuvant-induced arthritis(AA),so as to explore the underlying mechanism of moxibustion in the treatment of rheumatoid arthritis.Methods Forty-five SD rats were randomly divided into the blank control group,model group,moxibustion group,methotrexate group and rapamycin group,with 9 rats in each group.The rat model of AA was established by injecting Freund’s complete adjuvant.Rats in the moxibustion group received moxibustion treatment at“Zusanli”(ST36)and“Guanyuan”(CV4)for 20 min,once a day.The methotrexate group was given methotrexate intragastrically(0.35 mg/kg)twice a week.The rapamycin group was given rapamycin by intraperitoneal injection(1 mg/kg),once every other day.The toe volume of the left hind limb was measured by the toe volume measuring instrument after 3-day modeling and 3-week intervention respectively.The contents of interlukin(IL)-1 and tumor necrosis factor(TNF)-αin serum were detected by ELISA.The autophagosomes of synovial cells of the toe joint were observed under transmission electron microscope.The expressions of mammalian target of rapamycin(mTOR)C1,p-mTORC1,Caspase-3,Fas and FasL in synovial tissue were detected by Western blot.Results Under transmission electron microscope,the model group showed decreased autophagosomes in synovial tissues,but the moxibustion,methotrexate,and rapamycin groups showed increased autophagosomes.Compared with the blank control group,the toe volume,the contents of IL-1 and TNF-αin serum and the expression of p-mTORC1 protein in synovial tissue were significantly increased(P<0.01,P<0.001),while the expressions of Caspase-3,Fas and FasL proteins in synovial tissue were significantly decreased(P<0.05,P<0.01)in the model group.Compared with the model group,the toe volume,the contents of IL-1 and TNF-αin the serum,and expression of p-mTORC1 protein were significantly decreased(P<0.05,P<0.01,P<0.001)in the moxibustion group an

关 键 词：艾灸 类风湿性关节炎 滑膜细胞 自噬 细胞凋亡 

分 类 号：R245.81[医药卫生—针灸推拿学]