中药复方溃结灵抑制NF-κ#27963;化的Iκ#27867;素化机制研究

项目名称： 中药复方溃结灵抑制NF-κ#27963;化的Iκ#27867;素化机制研究

项目编号： No.30873421

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 杜群

作者单位： 广州中医药大学

项目金额： 30万元

中文摘要： 对溃疡性结肠炎起主要致病作用的细胞因子在转录水平受到NF-κ#30340;调控，Iκ#30340;泛素化是NF-κ#27963;化的必经环节。本研究采用TNBS灌肠制作UC大鼠模型，采用TNF-α12289;IL-1β35825;导caco-2细胞制备炎症细胞模型，观察溃结灵及其含药血清对两种模型NF-κ#27963;化的作用；并采用酶联免疫、PCR、免疫印迹等方法对NF-κ#27963;化的重要环节Iκ#945;、IKKβ12289;泛素系统的泛素、E1、E2、E3蛋白表达和基因表达进行了检测，以期探讨该方作用的深层次机理。整体动物实验结果表明，溃结灵能明显升高UC大鼠Iκ#945;的蛋白和基因表达水平，降低IKKβ34507;白表达，降低泛素、E1、E2、E3蛋白表达及降低泛素和E2基因表达。细胞实验结果表明，溃结灵含药血清能明显抑制TNF-α21644;IL-1β35825;导的caco-2炎症细胞NF-κP65蛋白DNA结合活性，提高Iκ#945;蛋白和基因表达，降低TNF-α35825;导的炎症细胞的IKKβ12289;泛素、E1、E2、E3蛋白表达和E3基因表达，降低IL-1β35825;导的炎症细胞的泛素、E1、E2、E3蛋白表达和IKKβ12289;E3的基因表达。提示抑制泛素系统功能是溃结灵抑制NF-κ#27963;化的部分作用机制。

中文关键词： 溃疡性结肠炎；溃结灵；核因子-κ#65307;Iκ#65307;泛素化

英文摘要： The cytokines which play a main pathogenic role in ulcerative colitis (UC) are regulated by Nuclear factor kappa B (NF-kB) on the transcription level while the ubiquitinylation of IkB is a key part of NF-KB activation.In this study,we use the TNBS clyster to establish the UC rat model and preparation of caco-2 inflammatory cells model induced by tumor necrosis factor-αNF αand interleukin 1βL 1βin order to observe function of KuiJieling and it's medicated serum on NF-kB activation of two kinds models. protein and mRNA expression of Iκalpha, IKK βubiquitin,ubE1,ubE2 and ubE3, which are the important section of NF-kB activation are detected by euzymelinked immunosorbent assay、PCR and immunoblotting. The results show that KuiJieling can obviously increase the protein and mRNA expression of Iκalpha in UC rat model, decrease the protein expression of IKK βubiquitin, ubE1，ubE2 and ubE3，it also can down regulate the mRNA expression of ubE2 and ubiquitin. The result of cell experiment show that KuiJieling medicated serum can obviously inhibit activity of NF-kB p65 protein combined with DNA in caco-2 inflammatory cells model induced by TNF-αnd IL-βincrease the protein and mRNA expression of Iκ#945;. decrease the protein expression of IKKβ12289;Ubiquitin、E1、E2、E3 of TNF-αnduced inflammatory cells and the mRNA expression of E3；At the same time，it could also down regulate the protein expression of Ubiquitin、E1、E2、E3 of IL-1βnduced inflammatory cells and the mRNA expression of IKKβ12289;E3. The results indicate that, ,inhibiting the ubiquitin-proteasome system is the part of the mechanism of KuiJieling on restraining NF-kB activation.

英文关键词： Ulcerative Colitis; KuiJieling; Nuclear factor kappa B (NF-kB); Iκ Ubiquitination