L-161982对实验性自身免疫性神经炎大鼠趋化因子表达的影响

发布时间：2018-05-29 07:21

本文选题：大鼠 + 实验性自身免疫性神经炎　；参考：《山东大学》2014年硕士论文

【摘要】：研究目的：吉兰-巴雷综合征(Guillain-Barre syndrome, GBS)是以周围神经髓鞘脱失和小血管炎性细胞浸润为病理特点的自身免疫性周围神经病,以肢体对称性弛缓性瘫痪为主要临床特征。实验性自身免疫性神经炎(experimental autoimmune neuritis,EAN)是国际公认的研究GBS的动物模型,是T辅助细胞1和T辅助细胞17(Thl/Th17)介导的自身免疫性疾病,以周围神经脱髓鞘和大量炎性细胞浸润为主要病理特点。这些浸润的炎性细胞主要包括单核巨噬细胞、淋巴细胞以及粒细胞,促炎性细胞因子如IFN-γ、TNF-α、NO及趋化因子MCP-1、MIP-1等在其发病中起关键作用。近年来研究认为前列腺素E2(Prostaglandin E2)是一种免疫激活剂,可能通过作用于EP4受体影响Thl和Th17这两个细胞亚群的分化来参与自身免疫性疾病的免疫炎症过程,由此我们推测PGE2-EP4受体拮抗剂对EAN具有治疗作用。在我们的前期研究中发现,应用PGE2-EP4拮抗剂L-161982干预EAN大鼠,可降低其坐骨神经中促炎性细胞因子IL-17及IFN-y的表达。基于趋化因子在GBS/EAN中的重要作用,本实验观察了PGE2-EP4拮抗剂L-161982对EAN大鼠趋化因子CXCL-12和MCP-1表达的影响。实验方法： 1.用周围神经鞘磷脂(BPM)免疫Lewis大鼠,制备EAN模型。 2.随机将21只EAN大鼠分为3组,治疗A组、治疗B组和对照组。两个治疗组均以剂量为5mg/kg的PGE2-EP4受体拮抗剂L-161982腹腔注射治疗。治疗A组于免疫阶段(免疫前1天至免疫后第8天)每日行L-161982处理,治疗B组于发病阶段(免疫后第5天至第14天)每日行L-161982处理。对照组在整个实验阶段每日给予相同体积的L-161982溶媒DMSO腹腔注射。 3.免疫后每日观察EAN大鼠的临床评分。于免疫后第15天处死EAN大鼠,取其坐骨神经,用免疫组化法检测坐骨神经中趋化因子CXCL-12及MCP-1的表达水平。 4.应用SPSS18.0软件对资料进行统计分析,数据以x±s表示。实验结果： 1.与对照组相比,不同阶段处理的两个治疗组均能延迟EAN的发病时间(P0.05),降低其高峰期临床评分(P0.05),减少坐骨神经中CXCL-12和MCP-1的表达(P0.05)。 2.两个治疗组间发病时间、高峰期临床评分以及趋化因子表达的比较,均可见显著差异(P0.05)。结论： 1.应用L-161982治疗可明显延迟EAN大鼠的发病时间,降低其高峰期临床评分,改善EAN大鼠的临床症状,说明其对周围神经自身免疫性疾病有治疗作用。 2.应用L-161982治疗可抑制EAN大鼠坐骨神经中趋化因子CXCL-12及MCP-1的表达,提示PGE2-EP4受体拮抗剂L-161982可通过抑制巨噬细胞和T细胞的迁移活化,减少CXCL-12和MCP-1的产生,从而抑制免疫炎症反应,这可能是L-161982的抗炎机制之一。 3.与发病阶段相比,免疫阶段应用PGE2-EP4受体拮抗剂L-161982更能显著延迟EAN大鼠的发病时间,降低其高峰期评分,减少坐骨神经中趋化因子CXCL-12和MCP-1的表达。因此,免疫阶段使用L-161982治疗效果更显著。
[Abstract]:Objectives of the study: Guillain-Barre syndromeGuillain-Barre syndrome (GBSs) is an autoimmune peripheral neuropathy characterized by peripheral nerve demyelination and small vascular inflammatory cell infiltration. The main clinical feature is limb symmetric flaccid paralysis. Experimental autoimmune neuritis (EAN) is an internationally recognized animal model for the study of GBS. It is an autoimmune disease mediated by T helper cell 1 and T helper cell 17 Thl / Th17. The main pathological features were demyelination of peripheral nerve and infiltration of inflammatory cells. These infiltrating inflammatory cells mainly include mononuclear macrophages, lymphocytes and granulocytes, pro-inflammatory cytokines such as IFN- 纬 -TNF- 伪 no and chemokine MCP-1 / MIP-1, which play a key role in its pathogenesis. In recent years, it has been suggested that prostaglandin E2(Prostaglandin E2 (PGE) is an immune activator, which may be involved in the immune inflammation of autoimmune diseases by affecting the differentiation of Thl and Th17 subsets by acting on EP4 receptors. Therefore, we speculate that PGE2-EP4 receptor antagonist has therapeutic effect on EAN. In our previous study, it was found that PGE2-EP4 antagonist L-161982 could reduce the expression of pro-inflammatory cytokines IL-17 and IFN-y in sciatic nerve of EAN rats. Based on the important role of chemokines in GBS/EAN, the effects of PGE2-EP4 antagonist L-161982 on the expression of chemokines CXCL-12 and MCP-1 in EAN rats were studied. Experimental methods: 1. Lewis rats were immunized with peripheral nerve sphingolipid (BPM) to establish EAN model. 2. Twenty-one EAN rats were randomly divided into three groups: group A, group B and control group. Both groups were treated by intraperitoneal injection of PGE2-EP4 receptor antagonist L-161982 (5mg/kg). Group A was treated daily with L-161982 during the immune phase (from 1 day before immunization to 8 days after immunization) and group B was treated daily with L-161982 at the onset stage (from the 5th to the 14th day after immunization). The control group was given intraperitoneal injection of L-16 1982 solvent DMSO daily during the whole phase of the experiment. 3. The clinical scores of EAN rats were observed daily after immunization. On the 15th day after immunization, EAN rats were killed and their sciatic nerves were removed. The expression of chemokine CXCL-12 and MCP-1 in sciatic nerve was detected by immunohistochemical method. 4. The data were analyzed by SPSS18.0 software, and the data were expressed as x 卤s. Experimental results: 1. Compared with the control group, the two treatment groups at different stages could delay the onset of EAN, decrease the peak clinical score and decrease the expression of CXCL-12 and MCP-1 in sciatic nerve. 2. There were significant differences in onset time, peak clinical score and chemokine expression between the two groups (P 0.05). Conclusion: 1. L-161982 treatment can significantly delay the onset of EAN rats, reduce its peak clinical score, improve the clinical symptoms of EAN rats, indicating that it has therapeutic effect on peripheral nerve autoimmune diseases. 2. L-161982 treatment could inhibit the expression of chemokine CXCL-12 and MCP-1 in sciatic nerve of EAN rats, suggesting that PGE2-EP4 receptor antagonist L-161982 could inhibit the production of CXCL-12 and MCP-1 by inhibiting the migration and activation of macrophages and T cells. This may be one of the anti-inflammatory mechanisms of L-161982. 3. PGE2-EP4 receptor antagonist L-161982 significantly delayed the onset of EAN rats, decreased its peak score, and reduced the expression of chemokines CXCL-12 and MCP-1 in sciatic nerve. Therefore, the effect of L-161982 was more significant in the immune phase.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R744.5

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