溃疡性结肠炎患者肠粘膜κ基因结合核因子的活化及抗炎药物的作用

目的：探讨溃疡性结肠炎患者肠粘膜κ基因结合核因子（NF－κB）的活化以及柳氮磺胺吡啶（SASP）和糖皮质激素对其的影响。方法：31例溃疡性结肠炎患者中17例使用过药物（SASP或SASP＋糖皮质激素）治疗,14例未用过任何与溃疡性结肠炎治疗相关的药物,11例同期结肠癌患者（取其癌旁正常组织（作为对照。采用凝胶电泳迁移率改变分析（EMSA）检测NF－κB DNA结合活性,Western蛋白印迹分析检测NF－κB p65蛋白表达情况,免疫组化方法原位检测肠粘膜组织中NF－κBp65蛋白的表达情况,荧光双标激光共聚焦显微镜确定表达NF－ Bp65的细胞类型。结果：溃疡性结肠炎患者肠粘膜NF－κBp65的表达和NF－ B DNA结合活性与对照组比较明显升高（P＜0．05）,且与炎症程度有关;溃疡性结肠炎患者肠粘膜巨噬细胞,T淋巴细胞,B淋巴细胞以及隐窝上皮细胞均有NF－κBP65的活化;糖皮质激素和SASP明显抑制NF－ κB的活性及表达,结论：NF－κB活性和表达的增加可能与溃疡性结肠炎的发生发展有关;糖皮质激素和SASP的抗炎作用可能是通过抑制NF－κ κB的活性实现的。NF－κ B可能为溃疡性结肠炎新药的研究与开发提供了一个有价值的靶标。

Activation of nuclear factor-kappaB and effects of anti-inflammatory treatment thereon in intestinal mucosa of patients with ulcerative colitis

OBJECTIVE: To investigate the activation and expression of nuclear factor-kappaB (NF-kappaB) and effects of anti-inflammatory treatment on NF-kappaB in the intestinal mucosa of patients with ulcerative colitis (UC). METHODS: Ten pieces of colon mucosal biopsy specimens were obtained from 31 cases with UC, 17 of which received sulphasalazine (SASP) or SASP plus glucocorticoid and 14 of which received no medication. Samples of normal mucosa around the lesion taken from 11 patients with colon cancer were used as controls. NF-kappaB DNA binding activity was evaluated by electrophoretic mobility shift assay. NF-kappaB p65 expression was determined by Western blot analysis and immunohistochemical staining with a NF-kappaB p65 antibody. The type of cells containing activated NF-kappaBp65 was identified by double immunofluorescence confocal laser scanning microscopy. RESULTS: The expression of NF-kappaB p65 and NF-kappaB DNA binding activity were significantly higher in patients with UC than in the control (P < 0.05), and were correlated with the degree of inflammation. The NF-kappaB expression was significantly stronger in the nuclei than in the cytoplasm in patients with UC without pharmacotherapy. The NF-kappaB expression in nuclei was significantly stronger in the group without pharmacotherapy than in the group with pharmacotherapy (P < 0.05). Only a few NF-kappaB p65 positive cells were seen in the controls. NF-kappaBp65 expression was found in all major subsets of mononuclear cells, including macrophages, B lymphocytes, T lymphocytes, and cryptal epithelial cells. CONCLUSION: The increased activation of NF-kappaB and increased expression of NF-kappaB may be involved in the pathogenesis of UC. Glucocorticoids and SASP strongly inhibited NF-kappaB activation and expression. The inhibition of NF-kappaB activation may be a central part of the anti-inflammatory action of glucocorticoids and SASP, which might represent an important pharmacological mechanism in treatment of patients with UC. NF-kappaB will be an important target for cytokine-based therapy of UC.