23-乙酰泽泻醇B抑制IgE/Ag介导的肥大细胞的激活和过敏反应
发布时间:2021-10-11 14:57
目的:本文利用小鼠骨髓来源肥大细胞(bone marrow-derived mast cells,BMMCs)、嗜碱性白血病细胞【rat basophilic leukemia(RBL),RBL-2H3】和人肥大细胞(human mast cell,HMC-1)对三萜类化合物23-乙酰泽泻醇B(alisol B 23-acetate,AB23A)的体内外抗炎作用及机制进行研究,为新型抗炎药物的开发提供了理论依据和实验基础。方法:1.利用MTT方法检测不同浓度AB23A在BMMCs、RBL-2H3和HMC-1中无细胞毒浓度范围。2.利用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)测定AB23A对免疫球蛋白E(immunoglobulin E,Ig E)/抗原(antigen,Ag)激活的BMMCs中白三烯C4(leukotriene C4,LTC4)合成、组胺(histamine)释放、白细胞介素-6(interleukin-6,IL-6)生成等的影响。利用免疫荧光法测定AB23A对激活BMMCs内Ca2+动员的影响。3.利用Weste...
【文章来源】:天津医科大学天津市 211工程院校
【文章页数】:83 页
【学位级别】:硕士
【部分图文】:
AB23A抑制IgE/Ag刺激的BMMCs中PLCγ的磷酸化、组胺释放和Ca2+动员
31图 3. AB23A 通过 Akt/IκB/NF-κB 通路抑制 IL-6 的产生和 COX-2 的表达。(a)AB23A 对 IgE/Ag 介导的 BMMCs 中 IL-6 产生的影响;(a) AB23A 对 IgE/Ag 介导的 BMMCs 中 COX-2 产生的影响;(c) AB23A 对 IgE/Ag 介导的 BMMCs 中Akt/IκB/NF-κB 中相关蛋白表达及磷酸化水平的影响;(d-e) 相关蛋白灰度统计。Figure 3. AB23A inhibits the production of IL-6 and the expression of COX-2through part of the Akt/IκB/NF-κB pathway. (a) IgE-sensitized BMMCs werepre-treated with AB23A or Bay 61-3606 for 1 h and stimulated with DNP-HSA for 6h. IL-6 were measured by ELISA. (b) IgE-sensitized BMMCs were pre-incubatedwith aspirin for 2 h to eliminate the activity of COX-1. After washing, the BMMCswere treated for 1 h with the AB23A or Bay 61-3606 and then stimulated withDNP-HSA for another 7 h. Cell lysates were immunoblotted with the antibody for
AB23A抑制Syk相关通路
本文编号:3430738
【文章来源】:天津医科大学天津市 211工程院校
【文章页数】:83 页
【学位级别】:硕士
【部分图文】:
AB23A抑制IgE/Ag刺激的BMMCs中PLCγ的磷酸化、组胺释放和Ca2+动员
31图 3. AB23A 通过 Akt/IκB/NF-κB 通路抑制 IL-6 的产生和 COX-2 的表达。(a)AB23A 对 IgE/Ag 介导的 BMMCs 中 IL-6 产生的影响;(a) AB23A 对 IgE/Ag 介导的 BMMCs 中 COX-2 产生的影响;(c) AB23A 对 IgE/Ag 介导的 BMMCs 中Akt/IκB/NF-κB 中相关蛋白表达及磷酸化水平的影响;(d-e) 相关蛋白灰度统计。Figure 3. AB23A inhibits the production of IL-6 and the expression of COX-2through part of the Akt/IκB/NF-κB pathway. (a) IgE-sensitized BMMCs werepre-treated with AB23A or Bay 61-3606 for 1 h and stimulated with DNP-HSA for 6h. IL-6 were measured by ELISA. (b) IgE-sensitized BMMCs were pre-incubatedwith aspirin for 2 h to eliminate the activity of COX-1. After washing, the BMMCswere treated for 1 h with the AB23A or Bay 61-3606 and then stimulated withDNP-HSA for another 7 h. Cell lysates were immunoblotted with the antibody for
AB23A抑制Syk相关通路
本文编号:3430738
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