艾灸对佐剂性关节炎大鼠脊髓中N-甲基-D天冬氨酸受体-一氧化氮-环鸟苷酸通路的影响  

Effect of moxibustion on inflammatory pain and N-methyl-D aspartic acid receptor-nitric oxide-cyclic GMP pathway in spinal cord of adjuvant arthritis rats

作　　者：彭传玉[1] 胡玲 吴子建 蔡荣林[1] 王洁 PENG Chuan-yu;HU Ling;WU Zi-jian;CAI Rong-lin;WANG Jie(College of Acupuncture-moxibustion and Tuina,Anhui University of Chinese Medicine,Hefei 230038,China;Institute of Acupuncture and Meridian,Anhui Academy of Traditional Chinese Medicine,Hefei 230038)

机构地区：[1]安徽中医药大学针灸推拿学院,合肥230038 [2]安徽省中医药科学院针灸经络研究所,合肥230038

出　　处：《针刺研究》2022年第3期250-255,共6页Acupuncture Research

基　　金：安徽高校自然科学研究重点项目(No.KJ2018A0296、KJ2017A300);国家重点基础研究发展计划项目(No.2015CB554504)。

摘　　要：目的:观察艾灸对佐剂性关节炎(AA)大鼠脊髓中N-甲基-D天冬氨酸受体-一氧化氮-环鸟苷酸(NMDA-NO-cGMP)通路的影响,探讨艾灸改善类风湿性关节炎(RA)炎性痛的作用机制。方法:SD大鼠随机分为正常组、模型组、艾灸组、艾灸+NMDA受体拮抗剂(AP-5)组和艾灸+NMDA受体激动剂(NMDA)组,每组20只。采用风寒湿环境因素结合右后足跖注射完全弗氏佐剂建立AA大鼠模型。艾灸组采用艾条悬灸“足三里”“肾俞”,两穴交替使用;艾灸+AP-5组和艾灸+NMDA组分别给予腹腔注射AP-5(0.7 mg·kg^(-1)·d^(-1))和NMDA(5 mg·kg^(-1)·d^(-1))后进行与艾灸组相同的干预;3组均每日1次,共干预15 d。造模前后及干预后测定大鼠右后足底机械痛阈(PWT);荧光定量PCR法和Western blot法检测脊髓组织中诱导型一氧化氮合酶(iNOS)mRNA和蛋白的表达;ELISA法检测脊髓组织中cGMP的含量;比色法和硝酸还原酶法分别检测脊髓组织中NOS的活性和NO的含量。结果:造模后,与正常组比较,其余各组右后足底PWT均显著降低(P<0.01)。干预后,与正常组比较,模型组右后足底PWT显著降低(P<0.01),脊髓组织中iNOS mRNA和蛋白的表达及cGMP、NO含量和NOS活性均明显升高(P<0.01)。与模型组比较,各干预组PWT明显升高(P<0.01),脊髓组织中iNOS mRNA和蛋白的表达及cGMP、NO含量和NOS活性均显著降低(P<0.01,P<0.05)。与艾灸组比较,艾灸+AP-5组PWT明显升高(P<0.01),脊髓组织中iNOS mRNA和蛋白的表达及NO含量均显著降低(P<0.01,P<0.05);艾灸+NMDA组PWT显著降低(P<0.01),脊髓组织中iNOS mRNA和蛋白的表达及cGMP、NO含量和NOS活性均明显升高(P<0.01)。结论:艾灸可改善AA大鼠的炎性痛反应,其作用机制可能与下调脊髓组织中NMDA-NO-cGMP通路的功能有关。Objective To observe the effect of moxibustion on pain and N-methyl-D aspartic acid receptor/nitric oxide/cyclic guanosine monophosphate(NMDA-NO-cGMP)signaling pathway in the spinal cord of rats with adjuvant arthritis(AA),so as to explore its underlying mechanisms in relieving inflammatory pain of rheumatoid arthritis(RA).Methods SD rats were randomly divided into normal,model,moxibustion(Moxi),Moxi+NMDA receptor antagonist AP-5(Moxi+AP-5)and Moxi+NMDA receptor agonist(NMDA)groups,with 20 rats in each group.The AA model was established by placing the rats in a wind,cold and damp environment for 12 h,once daily for 20 days and by injection of complete Freund’s adjuvant into the right hind paw.Rats of the three Moxi groups received ignited moxa-stick stimulation of“Zusanli”(ST36)and“Shenshu”(BL23)alternately for 20 min,once a day for 15 days.The Moxi+AP-5 group and Moxi+NMDA group received intraperitoneal injection of AP-5(0.7 mg·kg^(-1)·d^(-1))and NMDA(5 mg·kg^(-1)·d^(-1)),respectively,once a day,for a total of 15 days.Mechanical pain thres-hold(MPT)was measured before and after modeling and interventions.The spinal cord tissue was sampled for detecting the expression of iNOS mRNA and protein,content of cGMP and NO,and the activity of NOS by using fluorescence quantitative PCR,Western blot,ELISA,nitrate reductase method and colorimetric method,respectively.Results Before modeling,there was no significant difference in MPT among all the 5 groups(P>0.05).After modeling,the MPT was remarkably decreased(P<0.01),the expression levels of iNOS mRNA and protein,the contents of cGMP and NO,the activity of NOS were significantly increased in the model group relevant to the normal group(P<0.01).After the interventions,the MPT was obviously increased(P<0.01),while the expression levels of iNOS mRNA and protein,the contents of cGMP and NO,the activity of NOS were significantly down-regulated in the Moxi,Moxi-AP-5 and Moxi+NMDA groups(P<0.05,P<0.01).The effect of Moxi+AP-5 group was significantly superior to that

关 键 词：炎性痛 艾灸 脊髓 类风湿性关节炎 N-甲基-D天冬氨酸受体 一氧化氮 环鸟苷酸 

分 类 号：R245.81[医药卫生—针灸推拿学]