本文選題：布洛芬-2-芳基嗎啉乙酯衍生物 + 鞭毛蛋白��； 參考：《中南大學(xué)》2014年碩士論文

【摘要】：目的：通過鞭毛蛋白誘導(dǎo)的THP-1細(xì)胞株為炎癥模型,篩選出具有抗炎活性的布洛芬-2-芳基嗎啉乙酯衍生物及初步探討其是否通過抑制NF-κB活性發(fā)揮抗炎作用,為開發(fā)新的非甾體抗炎藥提供基礎(chǔ)的理論支持。 方法：分別用同一濃度的布洛芬、布洛芬-2-芳基嗎啉乙酯衍生物孵育THP-11h后,繼而用10ng/mL鞭毛蛋白刺激,測定各組THP-1細(xì)胞分泌TNF-α和IL-1p的能力,篩選出具有抗炎活性的布洛芬嗎啉乙酯衍生物。繼而測定NF-κB活化情況及NF-κB信號(hào)通路上游TLR5的表達(dá)水平。ELISA檢測細(xì)胞上清液中TNF-α和IL-1p的水平；Westernblotting檢測NF-κB p65蛋白和TLR5蛋白表達(dá)水平；EMSA檢測胞核內(nèi)NF-κB的DNA結(jié)合活性；RT-PCR檢測TLR5mRNA的表達(dá)水平。 結(jié)果： 1.與正常對(duì)照組相比,鞭毛蛋白孵育THP-1細(xì)胞株24h后,分泌TNF-α和IL-1β的能力、NF-κBp65蛋白及DNA結(jié)合活性、TLR5mRNA表達(dá)水平、THP-1細(xì)胞上TLR5蛋白表達(dá)水平有顯著差異(P0.05)。 2.與鞭毛蛋白組相比,布洛芬、布洛芬-2-芳基嗎啉乙酯衍生物A和B能下調(diào)TNF-α、IL-1β的水平,布洛芬-2-芳基嗎啉乙酯B下調(diào)作用更明顯(P0.01)。 3.與鞭毛蛋白組相比,布洛芬、布洛芬-2-芳基嗎啉乙酯A及B均能抑制鞭毛蛋白誘導(dǎo)的TLR5和NF-κBp65表達(dá)。 4.與鞭毛蛋白組相比,布洛芬-2-芳基嗎啉乙酯A和B不同程度上抑制核內(nèi)NF-κB的表達(dá)及DNA結(jié)合活性。 結(jié)論： 1.鞭毛蛋白可激活THP-1細(xì)胞TLR5/NF-κB信號(hào)通路進(jìn)而促進(jìn)炎癥因子分泌。 2.布洛芬-2-芳基嗎啉乙酯衍生物A和B可阻斷鞭毛蛋白激活TLR5/NF-κB信號(hào)通路的作用,從而抑制炎癥因子的分泌。 3.布洛芬-2-芳基嗎啉乙酯衍生物A和B可能具有COX-2/NF-κB雙重抑制活性,可為炎癥性疾病的基本治療提供理論支持。本實(shí)驗(yàn)：圖17幅,表格19個(gè),文獻(xiàn)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- 偽 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- 偽 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- 偽 and IL-1 尾, and ibuprofen -2-aryl morpholine ethyl ester B could down-regulate the level of TNF- 偽 -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.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R96

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本文編號(hào)：1916889