香兰素分子反向虚拟筛选及经吸嗅抗抑郁机制探讨

发布时间：2018-04-19 14:32

  本文选题：香兰素 + 反向虚拟筛选　； 参考：《安徽医科大学》2014年硕士论文

【摘要】：研究背景及目的 MDD的干预目前主要采用5-HT再摄取抑制剂（SSRI）等抗抑郁药物。在肯定这些新型抗抑郁药研究成果的同时，其疗效、毒副作用及给药途径等依然需要研究者的继续改进。香兰素分子具有奶香的气息，是被广泛使用的可食用香料。研究显示香兰素具有抗焦虑、抗菌、抗氧化等作用。自然界及人体内香兰素有哪些蛋白作用靶点？ 由于嗅觉通路与情感中枢杏仁核、海马等有广泛的纤维联系，这成为嗅觉通路调节情感中枢的理论基础，经嗅觉通路对神经系统进行干预的报道也越来越多。嗅觉通的给药途径起效迅速、无外周毒、副作用、途径方便。香兰素分子“令人愉快”的气味对MDD患者能否起到干预作用，，其机制是什么？ 本实验通过计算机反向虚拟筛选，探讨香兰素可能作用的蛋白晶体，并通过让MDD大鼠吸嗅含香兰素的气雾剂，研究香兰素吸嗅的抗抑郁作用及机制。 第一部分香兰素分子药用靶标计算机反向虚拟筛选 药物与蛋白质分子（受体）的结合类似于钥匙与锁的识别关系。为了探讨香兰素可能作用的受体蛋白，利用分子对接软件Surflex对sc-PDB蛋白数据库进行反向虚拟筛选，软件对筛选结果进行打分，打分结果大于8分的说明与靶标有较大的相关性，分值越高的蛋白与香兰素分子的相关性也就越高，这种虚拟筛选为香兰素的作用受体的研究及药用价值的开发提供了指导意义。 结果 筛选出来的大于8分的蛋白晶体为：羧乙基精氨酸β-内酰胺酶、金属β内酰胺酶、结合分支杆菌水杨酸合成酶、β内酰胺酶（IMP-1）。打分前20位的蛋白晶体见表3-1。筛选结果与香兰素抗菌、抗氧化作用的报道基本一致。 第二部分MDD大鼠吸嗅香兰素神经行为学观察 CUMS+孤养的方法建立大鼠MDD模型。根据干预的方式将建模成功后的大鼠分为：香兰素吸嗅组、氟西汀灌胃组、纯水吸嗅组，以及OBX+香兰素吸嗅组和假手术组（Sham）。每组大鼠给予相应的干预4w，观察4w后各组大鼠糖水消耗量、强迫游泳不动时间较干预前的变化。 结果 与干预前相比，干预4w后香兰素吸嗅组和氟西汀灌胃组大鼠FST不动时间、SCT糖水消耗有显著性差异；纯水吸嗅组和OBX+香兰素吸嗅组未见显著性差异。 第三部分香兰素吸嗅对大鼠脑内5-HT、DA、NE浓度的影响 神经行为学的观察结果显示，香兰素能经嗅觉通路改善大鼠的抑郁样行为。为了探索其作用机制，将大鼠麻醉处死后取脑，通过UPLC FLR （超高压液相色谱 荧光检测器）检测各组大鼠脑匀浆液内5 HT、 DA、 NE量，并通过蛋白试剂盒（BCA protein assay kits）检测各组大鼠脑匀浆液可溶性蛋白含量（Pr）作为内标。 结果 香兰素吸嗅组和氟西汀灌胃组大鼠脑匀浆液5-HT、DA的量显著高于纯水吸嗅组，而NE未见显著性差异。OBX+香兰素吸嗅组大鼠脑匀浆液5-HT、DA、NE均显著低于Sham组。 结论 1香兰素反向虚拟筛选的结果与其抑菌、抗氧化等报道一致 2香兰素吸嗅能改善SD大鼠抑郁样行为 3香兰素抗抑郁作用可能是提升了脑5-HT、DA所实现
[Abstract]:Background and purpose of study

At the same time , the therapeutic effect , toxic side effect and administration route of the novel antidepressants still need to be improved by the investigator . The vanillin molecule has the flavor of milk fragrance and is widely used edible spice . The research shows that vanillin has anti - anxiety , antibacterial , anti - oxidation , etc . The research shows that the vanillin has anti - anxiety , antibacterial , anti - oxidation and the like .

The olfactory pathway has a wide range of fiber connections , such as almond kernel , hippocampus and so on , which is the theoretical basis of olfactory pathway regulating the emotion center . The olfactory pathway has more and more reports on the nervous system intervention . The olfactory pathway has rapid effect , no peripheral toxicity , side effect and convenient route . The pleasant smell of vanillin molecule can interfere with the patients with MDD . What is the mechanism ?

This experiment was carried out by computer reverse virtual screening to investigate the possible protein crystals of vanillin , and to study the antidepressant effect and mechanism of Vanillin ' s absorption and sniffing by inhalation of Vanillin - containing aerosol in MDD rats .

Reverse virtual screening of the first part of vanillin molecular medicine target computer

In order to explore the possible receptor protein of vanillin , a molecular docking software Surflex was used to reverse virtual screening of sc - PDB protein database .

Results

The screened protein crystals were carboxyethylarginine . beta . - lactamase , metal beta - lactamase , combined mycobacterial salicylic acid synthetase , beta - lactamase ( IMP - 1 ) . The protein crystals of the first 20 positions were shown in Table 3 - 1 . The screening results were consistent with the reported anti - bacterial and anti - oxidation effects of vanillin .

Neurobehavioral Observation of Olfactory Vanillin in the Second Part of MDD Rats

The rats were divided into three groups : vanillin inhalation group , fluoxetine group , pure water absorption and sniffing group , OBX + vanillin inhalation group and sham operation group ( Sham ) .

Results

Compared with the prior intervention , there was a significant difference in SCT sugar and water consumption after the intervention of 4w after 4w treatment of vanillin inhalation group and fluxitin irrigation group .
There was no significant difference in the sniffing group of pure water and OBX + Vanillin .

Effects of the third part of vanillin on the concentration of 5 - HT , DA and NE in rat brain

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