PDE4抑制剂FCPR03对LPS诱导小鼠抑郁样行为的改善作用及其机制研究

发布时间：2018-01-26 12:27

本文关键词： FCPR03 PDE4抑制剂抑郁症脂多糖细胞因子神经炎症　出处：《南方医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:抑郁症是由多种因素引起的,以持续心境低落为主要临床特征的情感性精神障碍,是一个世界性的重大心理健康问题。近年来,随着现代生活节律的不断加快,社会压力的逐渐增大,抑郁症的发病率也呈逐年升高的趋势。许多的研究认为抑郁症的发病与中枢的炎症过程有着密切联系,同时越来越多的研究也表明炎症在抑郁症的发生和发展过程中具有重要作用,抑郁症和中枢炎症之间有着某种密切联系。大量研究显示PDE4抑制剂在体内外均具有显著的抗炎作用,同时还能够通过上调cAMP/PKA/CREB信号通路产生抗抑郁的作用,然而因具有严重的恶心呕吐不良反应,限制了其在临床上的进一步应用。FCPR03是本课题组自主设计并合成的新型PDE4抑制剂,前期的实验结果显示对PDE4具有很高的选择性。基于抑郁症的神经炎症假说,本课题利用LPS建立炎症诱导的小鼠抑郁症模型,评价FCPR03对LPS诱导的小鼠抑郁样行为的改善作用,同时利用LPS诱导BV-2小胶质细胞的活化作为体外神经炎症模型,探讨FCPR03的抗神经炎症作用及其机制研究,为寻找能够改善由炎症诱导抑郁症的药物提供理论基础。方法:(1)采用抑郁症行为绝望模型中悬尾和强迫游泳实验初步探讨FCPR03的抗抑郁作用。(2)建立LPS诱导小鼠抑郁样行为动物模型,对小鼠腹腔注射1.2mg/kg LPS作为炎症诱导的小鼠抑郁症模型,评价FCPR03对LPS诱导小鼠抑郁样行为的改善作用及其相关机制的研究。(3)利用1 μg/mlLPS诱导BV-2小胶质细胞活化模拟体外神经炎症模型,评价FCPR03的体外抗神经炎症作用。(4)通过观察氯胺酮/赛拉嗪联合诱导小鼠翻正反射从消失到恢复所需时间以及比格犬的致呕吐实验来评价FCPR03的致呕吐潜能。结果:(1)和正常组相比,小鼠腹腔注射不同剂量FCPR03 30 min后,在旷场实验中的水平得分和垂直得分均没有统计学差异;而在悬尾和强迫游泳实验中,小鼠不动时间显著降低,表明FCPR03具有潜在的抗抑郁作用。(2)对小鼠腹腔注射1.2mg/kgLPS24h后,小鼠体重显著降低,同时在悬尾和强迫游泳实验中小鼠不动时间显著增加,糖水偏嗜度明显降低,而连续7天灌胃给予FCPR03后,能够降低小鼠在行为学中的不动时间,增加小鼠的糖水偏嗜度,提示FCPR03可以改善LPS诱导的小鼠抑郁样行为。(3)进一步的研究发现,FCPR03可能是通过上调cAMP/PKA/CREB信号通路,增加皮层和海马内BDNF的表达,降低p38和JNK的磷酸化水平,抑制NF-κB的活化,从而抑制炎症因子的释放,抑制中枢的神经炎症,改善LPS诱导的小鼠抑郁样行为。(4)和正常组相比,FCPR03对赛拉嗪和氯胺酮联合麻醉后小鼠翻正反射恢复时间没有统计学差异,且不引起比格犬的呕吐反应,提示FCPR03具有较低的致呕吐潜能。结论:FCPR03能够上调cAMP/PKA/CREB信号通路,降低p38和JNK的磷酸化水平,抑制NF-κB的活化,抑制炎症因子的释放,降低中枢的神经炎症,进而改善LPS诱导的小鼠抑郁样行为。同时,FCPR03具有较低的致呕吐潜能,提示FCPR03是一个潜在的抗抑郁候选药物。
[Abstract]:Objective: depression is caused by many factors, with sustained low mood affective disorders as the main clinical features, is one of the major health problems of psychological. In recent years, with the development of modern life rhythm speeding up, increasing social pressure, depression incidence is increasing year by year. Many studies suggest that the inflammatory process and the incidence of central depression are closely related, but more and more studies show that inflammation plays an important role in the occurrence and development of depression, between depression and central inflammation have a close relationship. A large number of studies show that PDE4 inhibitors have significant anti-inflammatory effect in vivo, at the same time can also produce antidepressant effect through upregulation of cAMP/PKA/CREB signaling pathway, however, because of the serious adverse reaction of nausea and vomiting, limiting its clinical. The further application of.FCPR03 is the independent research group design and synthesis of novel PDE4 inhibitors, the experimental results show that the high selectivity of PDE4. The nerve inflammation hypothesis of depression based on LPS to establish the animal model of depression induced by inflammation of the subject, to evaluate the improving effect of FCPR03 on mice depression like behavior induced by LPS, and the LPS BV-2 induced the activation of microglia as inflammation model in vitro of FCPR03 nerve, anti neuroinflammatory effect and its mechanism of study can provide a theoretical basis for the improvement of drugs by inflammation induced depression. Methods: (1) the depression model of behavioral despair tail suspension and forced swimming tests to explore the antidepressant effect of FCPR03. (2) establish the LPS induced depression like behavior in the animal model of mice after intraperitoneal injection of 1.2mg/kg LPS as a mice model of depression induced by inflammation, Study on the effect evaluation of FCPR03 on LPS induced depression like behaviors of mice and its related mechanism. (3) BV-2 induced microglial activation in vitro by nerve inflammation model simulated 1 g/mlLPS, anti inflammation effect evaluation of FCPR03 nerve in vitro. (4) through the observation of ketamine / xylazine induced righting reflex disappeared from the recovery time and beagle dogs were used to evaluate the FCPR03 induced vomiting induced vomiting potential. Results: (1) compared with the normal group, different doses of FCPR03 mice by intraperitoneal injection of 30 min, there were no statistical differences in the open field test score level and vertical score; and in the tail suspension and forced swimming test in the real time of mice decreased significantly, showed that FCPR03 has a potential antidepressant effect. (2) in mice after intraperitoneal injection of 1.2mg/kgLPS24h, body weight of mice was significantly reduced, at the same time in the tail suspension and forced swimming tests in mice Significant increase in real time, sucrose preference is significantly reduced, and 7 consecutive days after intragastric administration of FCPR03 in mice, can reduce behavior in real time, increase in sucrose preference, suggesting that FCPR03 can improve the LPS induced depression like behavior. (3) further research found that FCPR03 may be mediated by cAMP/PKA/CREB signaling pathway, increased expression of cortex and hippocampus in BDNF, p38 and JNK decreased the phosphorylation level of NF-, inhibiting the activation of NF kappa B, thereby inhibiting the release of inflammatory cytokines, suppression of central nerve inflammation, improve LPS induced rat depression like behavior. (4) compared with the normal group. FCPR03 of xylazine and ketamine combined anesthesia after righting reflex recovery time was not statistically significant, and does not lead to vomiting in beagle dogs, suggesting that FCPR03 has lower potential induced vomiting. Conclusion: FCPR03 can increase cAMP/PKA/CREB signaling pathway P38 and JNK, decreased the phosphorylation level of NF-, inhibiting the activation of NF kappa B, inhibiting the release of inflammatory cytokines, reducing inflammation of the central nerve, and improve the LPS induced depression like behavior. At the same time, FCPR03 has lower induced vomiting potential, suggesting that FCPR03 is a potential anti depression drug candidate.

【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R965

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