新型布洛芬衍生物的抗炎活性筛选及机制初步研究

发布时间：2018-05-21 00:19

本文选题：布洛芬-2-芳基吗啉乙酯衍生物 + 鞭毛蛋白　；参考：《中南大学》2014年硕士论文

【摘要】：目的：通过鞭毛蛋白诱导的THP-1细胞株为炎症模型,筛选出具有抗炎活性的布洛芬-2-芳基吗啉乙酯衍生物及初步探讨其是否通过抑制NF-κB活性发挥抗炎作用,为开发新的非甾体抗炎药提供基础的理论支持。方法：分别用同一浓度的布洛芬、布洛芬-2-芳基吗啉乙酯衍生物孵育THP-11h后,继而用10ng/mL鞭毛蛋白刺激,测定各组THP-1细胞分泌TNF-α和IL-1p的能力,筛选出具有抗炎活性的布洛芬吗啉乙酯衍生物。继而测定NF-κB活化情况及NF-κB信号通路上游TLR5的表达水平。ELISA检测细胞上清液中TNF-α和IL-1p的水平；Westernblotting检测NF-κB p65蛋白和TLR5蛋白表达水平；EMSA检测胞核内NF-κB的DNA结合活性；RT-PCR检测TLR5mRNA的表达水平。结果： 1.与正常对照组相比,鞭毛蛋白孵育THP-1细胞株24h后,分泌TNF-α和IL-1β的能力、NF-κBp65蛋白及DNA结合活性、TLR5mRNA表达水平、THP-1细胞上TLR5蛋白表达水平有显著差异(P0.05)。 2.与鞭毛蛋白组相比,布洛芬、布洛芬-2-芳基吗啉乙酯衍生物A和B能下调TNF-α、IL-1β的水平,布洛芬-2-芳基吗啉乙酯B下调作用更明显(P0.01)。 3.与鞭毛蛋白组相比,布洛芬、布洛芬-2-芳基吗啉乙酯A及B均能抑制鞭毛蛋白诱导的TLR5和NF-κBp65表达。 4.与鞭毛蛋白组相比,布洛芬-2-芳基吗啉乙酯A和B不同程度上抑制核内NF-κB的表达及DNA结合活性。结论： 1.鞭毛蛋白可激活THP-1细胞TLR5/NF-κB信号通路进而促进炎症因子分泌。 2.布洛芬-2-芳基吗啉乙酯衍生物A和B可阻断鞭毛蛋白激活TLR5/NF-κB信号通路的作用,从而抑制炎症因子的分泌。 3.布洛芬-2-芳基吗啉乙酯衍生物A和B可能具有COX-2/NF-κB双重抑制活性,可为炎症性疾病的基本治疗提供理论支持。本实验：图17幅,表格19个,文献88篇。
[Abstract]:Aim: to screen ibuprofen 2-aryl morpholine ethyl ester derivatives with anti-inflammatory activity and to explore whether they play an anti-inflammatory role by inhibiting NF- 魏 B activity in THP-1 cell line induced by flagellin. To provide theoretical support for the development of new non-steroidal anti-inflammatory drugs. Methods: THP-11h was incubated with ibuprofen and ibuprofen 2-aryl morpholine ethyl ester derivatives at the same concentration, and then stimulated by 10ng/mL flagellin. The ability of THP-1 cells to secrete TNF- 伪 and IL-1p was measured. Ibuprofen morpholine ethyl ester derivatives with anti-inflammatory activity were screened out. Then the activation of NF- 魏 B and the expression level of TLR5 upstream of NF- 魏 B signaling pathway were determined. The levels of TNF- 伪 and IL-1p in the supernatant were detected by Elisa. The expression levels of NF- 魏 B p65 protein and TLR5 protein in the supernatant were detected by Western blotting. The DNA binding activity of NF- 魏 B in nucleus was detected by EMSA and the expression of TLR5mRNA was detected by RT-PCR. Results: 1. After incubated with flagellin for 24 hours, the ability of secreting TNF- 魏 Bp65 protein and DNA binding activity of TLR5 mRNA in THP-1 cells was significantly higher than that in normal control group. There was significant difference in the expression of TLR5 protein in THP-1 cells. 2. Compared with flagellin group, ibuprofen -2-aryl morpholine ethyl ester derivatives A and B could down-regulate the level of TNF- 伪 and IL-1 尾, and ibuprofen -2-aryl morpholine ethyl ester B could down-regulate the level of TNF- 伪 -N-IL-1 尾. 3. Compared with flagellin group, Ibuprofen -2-aryl morpholine ethyl ester A and B could inhibit the expression of TLR5 and NF- 魏 Bp65 induced by flagellin. 4. Compared with flagellin group, ibuprofen-2-aryl morpholine ethyl ester A and B inhibited the expression of NF- 魏 B and DNA binding activity in nuclei to varying degrees. Conclusion: 1. Flagellin activates the TLR 5 / NF- 魏 B signaling pathway in THP-1 cells and promotes the secretion of inflammatory factors. 2. Ibuprofen -2-arylmorpholinyl ethyl ester derivatives A and B blocked the activation of TLR5 / NF- 魏 B signaling pathway by flagellin, which inhibited the secretion of inflammatory factors. 3. Ibuprofen-2-aryl morpholine ethyl derivatives A and B may have dual inhibitory activity of COX-2 / NF- 魏 B, which may provide theoretical support for the basic treatment of inflammatory diseases. This experiment: 17 pictures, 19 tables, 88 references.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96

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