火麻仁木脂素酰胺的抗神经炎症活性研究

发布时间：2018-06-24 20:12

本文选题：火麻仁木脂素酰胺 + Grossamide　；参考：《山东大学》2017年硕士论文

【摘要】：目前,神经退行性疾病(阿尔茨海默病,帕金森病和多发性硬化等)的发病率越来越高并趋于低龄化。这些疾病的发生、发展与氧化损伤、免疫炎症、线粒体功能性障碍、兴奋性毒素和细胞凋亡等机制有关。研究发现在神经退行性疾病的发生、发展过程中,小胶质细胞介导的炎症相关反应起着非常重要作用。细胞性毒素、化学、物理刺激等均会激活小胶质细胞,并释放大量的炎症因子,加剧炎症反应。小胶质细胞诱导神经炎症的发生并活化很多相关信号通路,比如:Toll样受体,JAK/STAT,NF-κB和MAPK等。其中NF-κB是介导炎症发生的重要信号通路,NF-κB的激活会诱导产生大量的炎症细胞因子,炎症因子反过来刺激细胞,使细胞发生持续的激活,进而控制大量细胞因子的基因表达。在小胶质细胞激活过程中,NF-κB会与其他炎症相关通路共同作用,调节炎症反应。所以,发现有效的控制NF-κB信号通路活化的化合物将有望成为神经退行性相关疾病治疗的潜在方法。火麻仁提取物在神经保护方面具有较好的活性,本课题组前期对火麻仁进行了化学成分研究,获得了一系列木脂素酰胺类化合物。本论文通过建立脂多糖(LPS)刺激BV2小胶质细胞的神经炎症模型,对这些化合物进行了抗炎活性筛选,并对筛选出的活性化合物grossamide、cannabisin F是否抑制LPS引起的NF-κB信号通路的激活进行研究,以发现其通过NF-κB信号通路干预来治疗神经炎症相关疾病的潜力。研究结果表明grossamide表现出潜在的抗炎活性。ELISA及qRT-PCR结果显示grossamide可以抑制LPS诱导的BV2细胞的炎症因子IL-6和TNF-α的表达。Western blot和免疫荧光等检测结果显示grossamide可以抑制IκBα及NF-κBp65蛋白的磷酸化,阻止NF-κBp65表达入核,同时阻止了 NF-κB上游蛋白TLR4及MyD88的表达。说明grossamide可以通过抑制TLR4介导的NF-κB的信号通路起到保护小胶质细胞的作用。研究结果表明cannabisin F同样表现出抗炎能力。细胞内活性氧检测结果显示cannabisin F可以抑制BV2细胞产生ROS。Western blot结果显示cannabisin F可以促进SIRT1,Nrf-2及下游蛋白HO-1的表达,同时可以抑制IκBα及NF-κBp65蛋白的磷酸化表达。ELISA及RT-PCR的结果显示cannabisin F可以抑制LPS刺激BV2细胞产生的炎症因子IL-6和TNF-α的表达。说明cannabisin F可通过激活SIRT1/Nrf-2信号通路,抑制NF-κB信号通路起到神经保护作用。
[Abstract]:At present, the incidence of neurodegenerative diseases (Alzheimer's, Parkinson's and multiple sclerosis, etc.) is becoming higher and lower. The occurrence and development of these diseases are related to oxidative damage, immune inflammation, mitochondrial dysfunction, excitatory toxins, and apoptosis. In the course of development, microglia mediated inflammation related reactions play a very important role. Cytotoxin, chemistry, physical stimulation, etc. can activate microglia and release a large number of inflammatory factors to aggravate the inflammatory response. Microglia induces neuroinflammation and activates many related signaling pathways, such as Toll like receptor, JAK /STAT, NF- kappa B and MAPK, in which NF- kappa B is an important signaling pathway to mediate the occurrence of inflammation, the activation of NF- kappa B induces a large number of inflammatory cytokines, which in turn stimulate the cells to enable the cell to continue to activate, and then control the gene expression of a large number of cytokines. In the process of microglia activation, NF- kappa B will be associated with the others. The inflammation related pathway acts together to regulate the inflammatory response. Therefore, the discovery of effective compounds to control the activation of the NF- kappa B signaling pathway is expected to be a potential method for the treatment of neurodegenerative diseases. A series of lignan amide compounds were obtained. In this paper, the inflammatory models of BV2 microglia were stimulated by lipopolysaccharide (LPS), and the anti-inflammatory activity of these compounds was screened. The activated compound grossamide, whether cannabisin F inhibited the activation of NF- kappa B signaling pathway caused by LPS, was studied. The potential of the treatment of neuroinflammatory related diseases through the intervention of NF- kappa B signaling pathway is present. The results show that grossamide shows the potential anti-inflammatory activity.ELISA and qRT-PCR results show that grossamide can inhibit the expression of IL-6 and TNF- a of BV2 cells induced by LPS, and the results of.Western blot and immunofluorescence are shown. Amide can inhibit the phosphorylation of I kappa B alpha and NF- kappa Bp65 protein, prevent the expression of NF- kappa Bp65 into nucleus, and prevent the expression of TLR4 and MyD88 of the upstream protein of NF- kappa B. The results of intracellular active oxygen detection showed that cannabisin F could inhibit the production of ROS.Western blot by BV2 cells and that cannabisin F could promote the expression of SIRT1, Nrf-2 and downstream protein HO-1. The expression of inflammatory factors IL-6 and TNF- alpha produced by the cells indicates that cannabisin F can inhibit the neuroprotective effect of the NF- kappa B signaling pathway by activating the SIRT1/Nrf-2 signaling pathway.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285

【参考文献】