电针“足三里”“关元”穴对佐剂性关节炎大鼠滑膜细胞凋亡及相关凋亡蛋白表达的影响  

Effect of electroacupuncture of“Zusanli”(ST36)and“Guanyuan”(CV4)on apoptosis and expression of apoptosis-related proteins of synoviocytes in adjuvant-induced arthritis rats

作　　者：刘梨[1] 周巍 黎铭玉 周兰 张亮[2] 王文怡 龚志贤[1] 艾坤[2] LIU Li;ZHOU Wei;LI Ming-yu;ZHOU Lan;ZHANG Liang;WANG Wen-yi;GONG Zhi-xian;AI Kun(The First Affiliated Hospital/the First Clinical College 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)

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]湖南中医药大学护理学院,长沙410208

出　　处：《针刺研究》2022年第8期696-702,共7页Acupuncture Research

基　　金：国家自然科学基金青年基金项目(No.81804204);国家自然科学基金面上项目(No.82074565);湖南省自然科学基金项目(No.2020JJ5433)。

摘　　要：目的:观察电针“足三里”“关元”对佐剂性关节炎(AIA)大鼠滑膜组织细胞凋亡率及其他凋亡蛋白含量的影响,探讨电针通过死亡受体(Fas)凋亡途径调节滑膜细胞凋亡和抗炎治疗类风湿关节炎(RA)的机制。方法: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。选用弗氏完全佐剂注射法制备AIA大鼠模型。造模后第3天开始干预,甲氨蝶呤组按照0.35 mg/kg的剂量进行灌胃,每周2次;电针组予以电针“足三里”“关元”,每次20 min,每周6次;两组均干预4周。用足趾容积测量仪测量各组大鼠左后肢足趾容积,HE染色观察踝关节滑膜组织形态变化,ELISA法检测大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1(IL-1)含量,TUNEL染色观察踝关节滑膜细胞凋亡率,Western blot法检测踝关节滑膜组织中凋亡相关基因Fas、死亡受体配体(FasL)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)的蛋白表达变化。结果:与正常组比较,模型组大鼠同时点左足趾容积增加(P<0.01),血清IL-1、TNF-α含量升高(P<0.01),滑膜细胞凋亡率及滑膜组织Fas、FasL、Caspase-3蛋白表达降低(P<0.01)。与模型组比较,甲氨蝶呤组、电针组大鼠第17、24天足趾容积减小(P<0.01),血清IL-1、TNF-α含量降低(P<0.01),滑膜细胞凋亡率增加(P<0.01),甲氨蝶呤组滑膜组织Fas、FasL蛋白表达升高(P<0.01,P<0.05),电针组滑膜组织Fas、Caspase-3蛋白表达升高(P<0.05)。与甲氨蝶呤组比较,电针组滑膜组织FasL蛋白表达降低(P<0.05)。HE染色显示模型组大鼠滑膜组织滑膜衬里层增生明显,滑膜细胞排列紊乱,部分细胞出现水肿,有纤维组织增生,炎性细胞浸润;甲氨蝶呤组大鼠滑膜部分增厚,炎性细胞浸润较模型组减少;电针组大鼠滑膜形态较模型组显著改善,滑膜增生情况较模型组减轻,炎性细胞浸润减少。结论:电针“足三里”“关元”能增加AIA大鼠滑膜细胞凋�Objective To observe the effect of electroacupuncture(EA)of“Zusanli”(ST36)and“Guanyuan”(CV4)on the apoptosis rate of synoviocytes and protein expression of Fas,FasL and Caspase-3 in synovial tissue of adjuvant-induced arthritis(AIA)rats,so as to explore its mechanisms underlying improvement of rheumatoid arthritis(RA).Methods A total of 24 rats were randomly divided into normal,model,medication and EA groups,with 6 rats in each group.The AIA model was established by injection of complete Freund’s adjuvant(CFA,0.1 mL)into the left hindlimb paw.The rats in the medication group received intraperitoneal injection of 0.35 mg/kg of methotrexate,twice a week for 4 weeks.The rats in the EA group received EA stimulation of ST36 and CV4(20 Hz/50 Hz,1 mA)for 20 min,6 times a week for 4 weeks.The left hind paw volume was measured using a paw volume meter,and histopathological changes of synovial tissue were observed by light microscope after H.E.staining.The serum contents of tumor necrosis factor-α(TNF-α)and interleukin-1(IL-1)were measured by ELISA.The apoptosis of synoviocytes was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL),and the expression of apoptosis-associated proteins Fas,FasL and Caspase-3 in synovium were detected by Western blot.Results Compared with the normal group,the left hind paw volume from day 3 to 24 after administration of CFA,serum IL-1 and TNF-αcontents were significantly increased(P<0.01),while the expressions of Fas,FasL and Caspase-3 proteins and apoptotic rate of synoviocytes were significantly decreased in the model group(P<0.01).In comparison with the model group,the paw volume from day 17 to 24 after modeling,the serum IL-1 and TNF-αcontents were significantly reduced(P<0.01),while the apoptotic rate of synoviocytes,expressions of Fas protein in both medication and EA groups,Caspase-3 protein in the acupuncture group and FasL protein in the medication group were increased(P<0.01,P<0.05).Compared with the medication group,the expression of F

关 键 词：电针 类风湿关节炎 滑膜细胞凋亡 死亡受体 死亡受体配体 半胱氨酸天冬氨酸蛋白酶-3 

分 类 号：R245.2[医药卫生—针灸推拿学]