Toll样受体9和核因子-κBp65在大鼠溃疡性结肠炎模型结肠组织中的表达及其相关性的研究

Expression of toll like receptor 9 andNF-κBp65 in colonic mucosa of rats with ulcerative colitis

目的观察Toll样受体9（TLR9）、核因子-κBp65（NF-κBp65）在溃疡性结肠炎（ulcerativecolitis，UC）大鼠模型结肠粘膜中的表达情况，探讨二者在UC发病机制中的作用及其相关性。方法运用复合法（2，4-二硝基氯苯＋乙酸）制备细胞免疫反应性UC大鼠模型，造模成功后，观察和评估结肠黏膜的大体形态和组织学变化；应用免疫组化Elivision法检测UC模型组和正常对照组大鼠结肠粘膜中TLR9和NF-κBp65表达，并分析其与病理组织学分级的关系。结果UC大鼠结肠粘膜中TLR9阳性表达率明显高于正常对照（P〈0．01）。在UC大鼠结肠粘膜组织学分级I、Ⅱ、Ⅲ级中的TLR9表达分别为13．75±9．78、49．33±17．76、82．4±29．46。表达的TLR9也存在显著差异，其中TLR9在I级与Ⅲ级UC大鼠中的表达有显著差异（P〈0．01），而Ⅱ级与Ⅰ级、Ⅱ级与Ⅲ级UC大鼠表达的TLR9也存在显著差异（P〈0．05）。NF-κB；p65在UC大鼠结肠粘膜中的阳性表达率明显高于正常对照组（P〈0．01）。在UC组织学分级为Ⅰ、Ⅱ、Ⅲ级的三组大鼠结肠粘膜中，NF-κBp65的表达分别为8．75±7．18、41．83±11．99、68．4±26．76。其中，I级与Ⅲ级UC大鼠相比，NF-κBp65的表达有极显著差异（P〈0．01），比较Ⅱ级与Ⅰ级、Ⅱ级与Ⅲ级UC大鼠NF-κBp65的表达也有显著差异（P〈0．05）。UC大鼠结肠粘膜中TLR9表达与NF-κBp65表达呈显著正相关（rs=0．971，P〈0．01）。结论UC大鼠结肠粘膜中TLR9、NF-κBp65表达明显升高，且与疾病的组织学分级呈正相关，提示其可能与UC的发生、发展有关。TLR9和NF-κBp65在UC结肠粘膜中的表达呈显著正相关，表明NF-κBp65是TLR9信号通路活化的产物。

Objective To observe the expression of toll-like receptol9 （ TLR9 ） and Nuclear Factor-κBp65 （NF-κBp65）in ulcerative colitis（UC） rat model, and to investigate their role in UC pathogenesis. Methods UC rat model was made by the treatment of 2, 4-dinitrochlorobenzene and acetic acid. Colonic mucosa and histological changes were observed and assessed. The expression of TLR9 and NF-κBp65 in UC rats were measured by immunohistochemical Elivision, and their relationship with the histological stages of UC was assessed. Results TLR9 level was significantly higher in UC rats compared with the control（ P 〈 0.01 ）. The expressions of TLR9 were 13.75 ± 9.78 in UC histological Stage Ⅰ , 49.33 ± 17.76 in UC histological Stage Ⅱ and 82.4 ± 29.46 in UC histological Stage m The expression of TLR9 was significantly different between Stage Ⅰ and Stage 0283 （P 〈0.0l ）. Significant difference between Stage Ⅱ and Stage Ⅰ , Stage Ⅱand Stagem （P〈0.05） was also observed. NF-κBp65 level was significantly higher in UC group compared with the control group（ P 〈 0.01 ）. The expressions of NF-κBp65 were 8.75 ± 7.18 ira UC histological Stage Ⅰ , 41.83 ± 11.99 in UC histological Stage Ⅱ and 68.4 ± 26.76 in UC histological Stage m . The expression of NF-κBp65 was significantly different between Stage Ⅰ and Stage Ⅲ （ P 〈 0.01 ）. Significant difference between Stage Ⅱ and Stage Ⅰ , Stage Ⅱ and Stage Ⅱ （ P 〈 0.05 ） was also observed. The expressions of TLR9 and NF-κBp65 were positively correlated with UC group （ rs = 0.971, P 〈 0.01 ）. Conclusion The high expression levels of TLR9 and NF-κBp65 in UC suggest that they may be related to the occurrence and development of UC . The positive correlation of TLR9 and NF-κBp65 with UC show that NF-κBp65 is the consequence of TLR9 signaling pathway activation.