四甲基吡嗪干预实验性自身免疫性葡萄膜炎的作用及其机制的研究

发布时间：2018-03-29 21:01

本文选题：四甲基吡嗪　切入点：眼内炎症　出处：《重庆医科大学》2017年硕士论文

【摘要】：背景:葡萄膜炎在临床上是常见的多发致盲性眼部疾病,主要由感染和自身免疫异常所致,发病病程长,难治易复发。长期反复持续的炎症可导致患者的视力产生不可逆且永久性的损害。目前发现自身免疫是葡萄膜炎发生的最主要致病病因,临床治疗的药物及方案一直是眼科领域里的重要研究课题。目的:本实验探讨了四甲基吡嗪(Tetramethylpyrazine,TMP)对实验性自身免疫性葡萄膜炎(Experimental autoimmune uveitis,EAU)动物模型的抗炎作用及其潜在的机制。方法:在B10RIII小鼠腿部及尾基部皮下注射IRBP肽诱导EAU动物模型。预防阶段的处理:对B10RIII小鼠分别使用TMP或vehicle腹腔注射3天后开始皮下注射IRBP制作EAU模型,再分别每天连续使用TMP或vehicle腹腔注射14天。效应阶段的处理:对B10RIII小鼠皮下注射IRBP制作EAU模型后第8天开始分别每天连续使用TMP或vehicle腹腔注射直至第14天。观察每组小鼠前房炎症再分别进行临床评分和组织学评分,评估EAU小鼠眼内炎症的严重情况,检测小鼠视网膜功能状态。提取每组小鼠视网膜的m RNA样本,并使用real-time PCR分别检查炎性因子IL-6、TNF-α、IL-1β、MCP-1,抗炎因子IL-10以及T细胞相关的因子IFN-γ和IL-17的表达变化。采用Western blotting检测p-STAT3和p-STAT4的蛋白表达水平。结果:在预防和效应这两个阶段中,TMP显著减轻EAU小鼠的前房及后房炎症,抑制IL-6、TNF-α、IL-1β、MCP-1、IFN-γ和IL-17的m RNA表达水平,并提高IL-10的m RNA水平。同时,经TMP治疗后的EAU小鼠ERG中a波和b波振幅明显升高,且p-STAT3和p-STAT4的蛋白表达水平明显下调。结论:全身应用TMP可明显减轻EAU小鼠的眼内炎症和视网膜功能障碍。这种抗炎作用与抑制STAT3和STAT4通路的活化有关。因此TMP可能成为治疗葡萄膜炎的新方案。
[Abstract]:Background: uveitis is a common ocular disease with multiple blindness in clinic. It is mainly caused by infection and autoimmune abnormality, and the course of the disease is long. Persistent inflammation can lead to irreversible and permanent visual impairment. Autoimmunity is now found to be the leading cause of uveitis. The drugs and protocols of clinical treatment have been an important research topic in ophthalmology. Objective: to investigate the anti-inflammatory effect and potential of Tetramethylpyrazine tetramethylpyrazine (TMP) on experimental experimental autoimmune uveitis (EAU) animal model. Methods: EAU animal model was induced by subcutaneous injection of IRBP peptide into the legs and base of tail of B10RIII mice. During the prevention stage, B10RIII mice were injected TMP or vehicle intraperitoneally for 3 days to make EAU model. The treatment of the effect stage: on the 8th day after the B10RIII mice were subcutaneously injected with IRBP to make EAU model, TMP or vehicle were injected intraperitoneally until the 14th day, respectively. The clinical and histological scores of anterior chamber inflammation were evaluated in each group. To evaluate the severity of intraocular inflammation in EAU mice, to detect the retinal function of mice, and to extract m RNA samples from the retina of each group. Real-time PCR was used to detect the expression of IL-6TNF- 伪 -IL-1 尾, anti-inflammatory factor IL-10 and T-cell related factors IFN- 纬 and IL-17, respectively. Western blotting was used to detect the expression of p-STAT3 and p-STAT4. Results: in the two stages of prevention and effect, the expression of p-STAT3 and p-STAT4 was detected. TMP significantly alleviated anterior chamber and posterior chamber inflammation in EAU mice. It inhibited the expression of m RNA in MCP-1tIFN- 纬 and IL-17, and increased the m RNA level of IL-10 in IL-6 TNF- 伪 -IL-1 尾. At the same time, the amplitudes of a and b waves in ERG of EAU mice treated with TMP were significantly increased. The protein expression of p-STAT3 and p-STAT4 was significantly down-regulated. Conclusion: systemic application of TMP can significantly reduce the intraocular inflammation and retinal dysfunction in EAU mice. This anti-inflammatory effect is related to the inhibition of the activation of STAT3 and STAT4 pathways. It can be a new treatment for uveitis.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R773.9

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