谷氨酸能信号在类风湿关节炎中的作用及其机制的研究

发布时间：2019-06-25 14:39

【摘要】：目的：建立大鼠胶原诱导型关节炎(Collagen-induced arthritis, CIA)模型,以离子型谷氨酸受体为研究靶点,分别应用受体拮抗剂MK-801及NBQX,观察不同受体拮抗剂对大鼠行为学的影响；观察应用拮抗剂后血清及组织液中COX-2, JAK3等表达的下调及上调；检测类风湿关节炎(Rheumatoid arthritis, RA)患者的血清谷氨酸水平,分析推断血液中谷氨酸浓度升高能否做为早期RA骨破坏的独立危险因素,从而研究探讨谷氨酸能信号通路在类风湿关节炎中的相关作用。方法：以Wistar大鼠作为实验对象,将大鼠随机分为6组,每组6只。第一组,空白对照组；第二组,CIA组；第三组,CIA+MK-801组；第四组,CIA+NBQX组；第五组,CIA+MK-801+NBQX组；第六组,8.7%DMS0+CIA组。观察研究CIA大鼠在注射不同拮抗剂后行为学改变,行为学评估内容包括触痛情况——使用机械性缩足阈值来评价以及肿胀情况的变化,肿胀情况使用排水法测足容积来代表。然后由大鼠心脏采血,并取组织匀浆液,检测血清及组织液中在拮抗剂使用后COX-2, JAK3等表达的下调及上调情况。同时,收集类风湿关节炎患者及健康人的血标本,分别检测其血清谷氨酸浓度,分析探讨其与抗CCP抗体,ESR,以及X线、B超分级等的关系。结果：对CIA大鼠分别注射不同的谷氨酸受体拮抗剂后,单独使用MK-801或NBQX以及联合使用这两种拮抗剂后均能表现出很好的恢复机械性缩足阈值(PWT)即镇痛作用。且镇痛作用持续大于24h,在镇痛作用方面,NBQX比MK-801表现更优。但无论是单独还是联合使用这两种拮抗剂,均不能改变CIA大鼠的足跖肿胀。NMDA受体拮抗剂MK-801可以降低CIA大鼠COX-2的表达,而对于Jak3,无论是单独亦或是联合使用此两种拮抗剂,对于CIA所致的Jak3表达增高情况均不能产生影响。谷氨酸浓度在类风湿关节炎患者与正常对照之间有显著统计学差异(P0.01),血清谷氨酸浓度在RA患者,与B超分级呈显著负相关(P0.01)。结论：谷氨酸受体拮抗剂MK-801及NBQX均能很好的减轻疼痛,NBQX比MK-801表现更优,但均对肿胀无效；NMDA受体与COX-2炎症通路有一定的交叉作用,而对于Jak3目前由该实验尚不能发现其与谷氨酸信号通路存在交叉作用；谷氨酸浓度与早期骨侵蚀有一定关系,可用作RA早期诊断的指标,其在类风湿关节炎中的检测中具有科研价值及临床应用价值。
[Abstract]:Objective: to establish a rat model of collagen-induced arthritis (Collagen-induced arthritis, CIA). The effects of different receptor antagonists on behavior of rats were observed by MK-801 and NBQX, respectively, and the expression of COX-2, JAK3 in serum and tissue fluid was down-regulated and upregulated after the application of antagonists. The serum glutamic acid level was measured in patients with rheumatoid arthritis (Rheumatoid arthritis, RA). It was analyzed whether the increase of glutamic acid concentration in blood could be an independent risk factor for early Ra bone destruction, so as to explore the related role of glutamic acid signaling pathway in rheumatoid arthritis. Methods: Wistar rats were randomly divided into 6 groups with 6 rats in each group. The first group, blank control group, the second group, CIA group, the third group, CIA MK-801 group, the fourth group, CIA NBQX group, the fifth group, CIA MK-801 NBQX group, the sixth group, 8.7%DMS0 CIA group. The behavioral changes of CIA rats after injection of different antagonists were observed and studied. the behavioral assessment included tenderness, which was evaluated by mechanical foot contraction threshold and the changes of swelling, and the swelling was represented by foot volume measured by drainage method. Then the blood was collected from the rat heart and the tissue homogenate was taken to detect the down-regulation and up-regulation of the expression of COX-2, JAK3 in serum and tissue fluid after the use of antagonists. At the same time, the blood samples of patients with rheumatoid arthritis and healthy people were collected to detect the serum glutamic acid concentration, and the relationship with anti-CCP antibody, ESR, X-ray, B-ultrasound grade and so on was analyzed and discussed. Results: after injection of different glutamate receptor antagonists into CIA rats, MK-801 or NBQX alone and combined use of these two antagonists could restore the analgesic effect of mechanical foot contraction threshold (PWT). The analgesic effect of NBQX was more than 24 hours, and NBQX was better than MK-801 in analgesic effect. However, either alone or in combination with these two antagonists could not change the paw swelling of CIA rats. NMDA receptor antagonist MK-801 could decrease the expression of COX-2 in CIA rats, but had no effect on the increase of Jak3 expression induced by CIA either alone or in combination with these two antagonists. The concentration of glutamic acid in patients with rheumatoid arthritis was significantly different from that in normal controls (P 0.01). The concentration of glutamic acid in serum was negatively correlated with B-ultrasound grade in patients with RA (P 0.01). Conclusion: glutamate receptor antagonists MK-801 and NBQX can relieve pain, NBQX is better than MK-801, but both of them have no effect on swelling. NMDA receptor and COX-2 inflammatory pathway have certain cross effect, but for Jak3, it can not be found that there is a cross effect with glutamic acid signaling pathway. Glutamic acid concentration is related to early bone erosion and can be used as an index for early diagnosis of RA. Glutamic acid concentration has scientific research value and clinical application value in the detection of rheumatoid arthritis.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.22

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