目的：观察核因子NF-κB P65信号通路介导细胞因子IL-23、趋化因子巨噬细胞炎症蛋白-3α（CCL20）及其受体CCR6表达的影响，及健脾方防治三硝基苯磺酸（TNBS）诱导的克罗恩病中相关作用机制。方法：24只清洁级雄性SD大鼠随机分为：①正常对照组（NG，n=8）：从造模第2周起双蒸水灌胃，每日1次，灌胃量按0.1 mL?kg^-1，共3周；②TNBS模型组（MG，n=8）：从第1周开始TNBS灌肠，灌肠剂量（mL）=体质量（g）×0.003ml?g^-1，每周1次，造模持续共4周；③模型+健脾方防治组（JP，n=8）：从造模第2周开始，在TNBS灌肠后的第2天予以中药健脾方灌胃治疗，灌胃量按0.1mL?kg^-1，每日1次，灌胃持续3周；采用结肠组织学损伤评分及HE组织病理的方法观察健脾方的疗效，免疫荧光、ELISA、原位杂交等技术观察各组大鼠结肠黏膜中NF-κB P65、IL-23、CCL20、CCR6蛋白与基因的表达差异；结果：免疫荧光和原位杂交结果显示，CCL20及其受体CCR6在正常组大鼠结肠中呈低表达，在模型组中呈高表达状态（P<0.05）。健脾方可显著降低大鼠结肠趋化因子CCL20及其受体CCR6的表达水平（P<0.05）。正常组大鼠结肠中NF-κB P65呈低表达，在模型组中呈高表达状态（P<0.05），健脾方可显著降低大鼠结肠NF-κB P65表达水平（P<0.05）。ELISA结果显示IL-23在正常组大鼠结肠中呈低表达，在模型组中呈高表达状态（P<0.05）。健脾方可显著降低大鼠结肠细胞因子IL-23的表达水平（P<0.05）。结论：健脾方可减轻TNBS诱导的CD大鼠结肠炎症程度，其潜在的机制与抑制NF-κB P65、IL-23和CCL20/CCR6的表达有关。

This study was aimed to observe the effect of nuclear factor NF-κB P65 signaling pathway on cytokine IL-23,chemokine macrophage inflammatory protein-3(CCL20)and its receptor CCR6 expression,and related mechanisms of spleen-invigorating prescription in the prevention and treatment of Crohn’s disease(CD)induced by trinitrobenzene sulfonic acid(TNBS).A total of 24 male SD rats were randomly divided into:(1)normal control group(NG,n=8):rats in this group were given distilled water from the second week,once a day,the amount of gastric lavage according to 0.1 mL?kg^-1,a total of 3 weeks;(2)model of TNBS group(MG,n=8):rats in this group were given TNBS enema from the first week,enema dose(mL)=body weight(g)×0.003ml?g^-1,once a week,the molding lasted a total of 4 weeks;(3)model of TNBS and spleen-invigorating prescription treatment group (JP,n=8):rats in this group were treated with traditional Chinese medicine(TCM)of spleen-invigorating prescription in the second day from the beginning of the second week after TNBS enema,the amount of gastric lavage treatment according to 0.1mL?kg^-1,once a day by gavage for 3 weeks.The curative effect of the method of the colon tissue damage and the pathology of HE were used to observe the curative effect of the spleen-invigorating prescription.Immunofluorescence,ELISA and in situ hybridization were used to observe the expression of NF-κB P65,IL-23,CCL20,CCR6 protein and gene in colon mucosa of rats in each group.The results of immunofluorescence and in situ hybridization showed that CCL20 and CCR6 in the normal group rat’s colon were less expressed,while in the model group showed high expression(P<0.05).Spleen-invigorating prescription can significantly reduce the expression of CCL20 and its receptor CCR6 in rat’s colon(P<0.05).NF-κB P65 in normal rat’s colon showed low expression,while in the model group showed high expression(P<0.05).And spleen-invigorating prescription can significantly reduce rat’s colon NF-κB P65 expression level(P<0.05).ELISA results showed that the expression of IL-23 was low in the colon of rats in the normal group,while in the model group showed high expression(P<0.05).Spleen-invigorating prescription can significantly reduce the rat colonic cytokine IL-23 expression levels(P<0.05).It was concluded that spleen-invigorating prescription can reduce the degree of colitis in CD rats induced by TNBS.Its underlying mechanism was related to the inhibition of the NF-κB P65 signaling pathway,and then affect the expression of IL-23 and CCL20/CCR6.

上海市教委优秀青年教师科研基金（zzszy12011）：克罗恩病模型制备及清热健脾方调节作用研究，负责人：吴璐一；上海中医药大学上海市针灸推拿学重点学科科研项目（JZ2012019）：灸药结合治疗克罗恩病的研究，负责人：吴璐一。