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IL-17对艾灸调控实验性RA大鼠RORγ-t、IκBζ信号转导因子含量影响的研究

发表时间:2016-12-15  浏览量:1356  下载量:280
全部作者: 高梓珊,甘君学,郭江燕,傅淑平,甘可
作者单位: 南京中医药大学第二临床医学院;南京中医药大学第一临床医学院
摘 要: 目的:探索艾灸治疗类风湿性关节炎(rheumatoid arthritis,RA)抗炎效应的白介素17(interleukin 17,IL-17)信号转导机制。方法:采用雄性SD大鼠66只,随机分为空白组、空白艾灸组、RA模型组(模型组)、RA模型艾灸组(艾灸组)、RA模型+IL-17沉默艾灸组、RA模型+IL-17过表达艾灸组、RA模型+IL-17沉默组、RA模型+IL-17过表达组。前6组每组各9只,后2组每组各6只。大鼠右后足垫部皮内注射完全弗氏佐剂0.15 mL进行RA造模。造模后采用慢病毒注射建立IL-17基因沉默、IL-17基因过表达模型。造模7 d后,艾灸大鼠“足三里”、“肾俞”两穴各5壮,每日1次,治疗18 d. 测量大鼠造模前后、艾灸治疗后足跖肿胀度;采用ELISA、Western blotting法检测大鼠血清IL-17、踝关节孤核受体(retinoid related orphan receptor gamma-t,RORγ-t)、核因子κB抑制因子ζ(recombinant inhibitory subunit of NF kappa B zeta,IκBζ)含量。结果:造模7 d后,大鼠右后足肿胀度明显高于空白组(P<0.01),艾灸后艾灸组鼠右后足肿胀度显著低于模型组(P<0.01)。在进行IL-17慢病毒注射之后,IL-17过表达组与模型组相比,IL-17含量明显上升,差异有统计学意义(P<0.05);IL-17沉默组与模型组相比,IL-17含量明显下降,差异有统计学意义(P<0.05)。艾灸治疗18 d后,艾灸组与模型组相比,艾灸能明显降低完全弗氏佐剂造模形成的佐剂性关节炎(adjuvant arthritis,AA)大鼠显著升高的IL-17、RORγ-t含量(P<0.05);艾灸能明显升高AA大鼠显著降低的IκBζ含量(P<0.01)。艾灸后IL-17过艾组与艾灸组相比,血清IL-17、转录因子RORγ-t含量明显上升(P<0.05);艾灸后IL-17沉艾组IL-17、RORγ-t含量明显下降(P<0.01)。艾灸后IL-17过艾组、IL-17沉艾组大鼠IκBζ含量与艾灸组相比无显著性差异。结论:艾灸可能通过调控IL-17,从而调节RA大鼠踝关节转录因子RORγ-t的含量,与IκBζ一起对RA发挥抗炎作用。
关 键 词: 针灸学;艾灸;类风湿性关节炎;白介素17;RORγ-t;IκBζ;抗炎
Title: Research on the effects of IL-17 on the contents of RORγ-t、IκBζ signal transduction factors in moxibustion regulation experimental RA rats
Author: GAO Zishan, GAN Junxue, GUO Jiangyan, FU Shuping, GAN Ke
Organization: Second School of Clinical Medicine, Nanjing University of Chinese Medicine; First School of Clinical Medicine, Nanjing University of Chinese Medicine
Abstract: Objective: To explore the interleukin17 (IL-17) signaling mechanism regarding to anti-inflammation effect of moxibustion in rheumatoid arthritis (RA) rats. Methods: 66 male SD rats were randomly assigned to control group, control+moxa group, RA model group (model), RA model moxa group (moxa), IL-17 knockdown+moxa group, IL-17 overexpression+moxa group, IL-17 knockdown group, IL-17 overexpression group. In the former 6 groups, each group has 9 SD rats, while 6 SD rats each in the latter two groups. 0.15 mL complete Freund’s adjuvant were injected into the right paw of SD rats for establishing RA model and using lentivirus injection to establish IL-17 knockdown/overexpression models. 18 days moxibustion treatments were conducted after 7 days injection. The treatment was that 5 times everyday moxibustion in each “Zusanli” and “Shenyu” points in SD rats. Changes of swelling degrees in RA rats’ right posterior foot before and after the model establishing, and then the moxibustion treatment were measured. Levels of IL-17, retinoid related orphan receptor gamma-t (RORγ-t) and recombinant inhibitory subunit of NF kappa B zeta (IκBζ) were measured using ELISA and Western blotting. Results: After 7 days injection, the swelling degree of RA rats’ right posterior foot was significantly higher than control group (P<0.01). After moxibustion treatment, swelling degree became significantly lower than model group (P<0.01). After IL-17 injection, IL-17 concentration were obviously higher in the IL-17 overexpression group than in the model group (P<0.05), while the concentrration of IL-17 got lower in IL-17 knockdown group than model group (P<0.05). After 18 days moxibustion treatment, comparing to model group, moxa group could significantly decrease the IL-17 and RORγ-t concentration that were increased by adjuvant arthritis (AA) rats after establishing complete Freund’s adjuvant model (P<0.05), while increase the IκBζ level (P<0.01). After moxibustion treatment, levels of IL-17 and RORγ-t in the serum of IL-17 overexpression+moxa group were increased than in moxa group (P<0.05); while these levels drop in the IL-17 knockdown+moxa group (P<0.01); however, there had shown no significant change in the IκBζ level among overexpression, knockdown and moxa groups. Conculsion: Through regulating IL-17 level, moxibustion treatment could have played a role in adjusting the RORγ-t and IκBζ concentration in RA rats’ ankle joint, thus demonstrated a cooperative anti-arthritis effect.
Key words: acupuncture; moxibustion; rheumatoid arthritis; interleukin 17; RORγ-t; IκBζ; anti-inflammation
发表期数: 2016年12月第23期
引用格式: 高梓珊,甘君学,郭江燕,等. IL-17对艾灸调控实验性RA大鼠RORγ-t、IκBζ信号转导因子含量影响的研究[J]. 中国科技论文在线精品论文,2016,9(23):2479-2488.
 
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