Göke Michael N, Schneider Maren, Beil Winfried, Manns Michael P
Division of Gastroenterology, Hepatology and Endocrinology, Medizinische Hochschule Hannover, Carl-Neuberg-Strasse 1, 30625, Hannover, Germany.
Regul Pept. 2002 May 30;105(3):203-14. doi: 10.1016/s0167-0115(02)00023-x.
Glucocorticoids are effective agents in the management of inflammatory bowel diseases. However, information about their effects on repair mechanisms of the intestinal epithelium is incomplete.Therefore, the aim was to analyse in vitro effects of glucocorticoids on proliferation, restitution, and apoptosis as well as their effects on activity and expression of nuclear factor (NF)-kappaB, a known regulator of apoptosis and inflammation, in intestinal epithelial cells.Non-transformed rat jejunum epithelial cells (IEC-6) were cultured in the presence and absence of various concentrations of prednisolone and budesonide. IEC-6 cell proliferation was assessed by [3H]-thymidine incorporation. Restitution was analysed by an IEC-6 in vitro assay. Apoptosis was evaluated by ELISA and fluorescence microscopy. DNA binding activity and nuclear expression of NF-kappaB was determined by electrophoretic mobility shift assays and Western blotting, respectively. Prednisolone and budesonide stimulated IEC-6 cell proliferation at low to medium pharmacologic concentrations (prednisolone: 10(-9) to 10(-6) M; budesonide: 10(-11) to 10(-8) M). In contrast, high concentrations (>5 x 10(-5) M) had inhibitory effects on proliferation. 10(-7) M prednisolone and 10(-8) M budesonide increased restitution of IEC-6 cells, whereas high concentrations (10(-4) M) of prednisolone and budesonide decreased restitution. Apoptosis of IEC-6 cells was substantially enhanced by 10(-4) M budesonide; apoptosis was slightly increased by the highest prednisolone concentration used (5 x 10(-4) M). Furthermore, both glucocorticoids inhibited DNA binding activity and nuclear NF-kappaB expression in IEC-6 cells in a dose- and time-dependent fashion. In conclusion, prednisolone and budesonide modulate repair mechanisms of intestinal epithelial cells in vitro in a dose-dependent manner and profoundly modulate the inflammatory regulator NF-kappaB.
糖皮质激素是治疗炎症性肠病的有效药物。然而，关于它们对肠上皮修复机制影响的信息并不完整。因此，本研究旨在分析糖皮质激素对肠上皮细胞增殖、修复和凋亡的体外影响，以及它们对已知的凋亡和炎症调节因子核因子（NF）-κB活性和表达的影响。将未转化的大鼠空肠上皮细胞（IEC-6）在存在和不存在不同浓度泼尼松龙和布地奈德的情况下进行培养。通过[3H]-胸腺嘧啶核苷掺入法评估IEC-6细胞增殖。通过IEC-6体外试验分析修复情况。通过酶联免疫吸附测定（ELISA）和荧光显微镜评估凋亡。分别通过电泳迁移率变动分析和蛋白质免疫印迹法测定NF-κB的DNA结合活性和核表达。泼尼松龙和布地奈德在低至中等药理浓度（泼尼松龙：10(-9)至10(-6)M；布地奈德：10(-11)至10(-8)M）时刺激IEC-6细胞增殖。相反，高浓度（>5×10(-5)M）对增殖有抑制作用。10(-7)M泼尼松龙和10(-8)M布地奈德可增加IEC-6细胞的修复，而高浓度（10(-4)M）的泼尼松龙和布地奈德则降低修复。10(-4)M布地奈德可显著增强IEC-6细胞的凋亡；所用最高浓度的泼尼松龙（5×10(-4)M）可使凋亡略有增加。此外，两种糖皮质激素均以剂量和时间依赖性方式抑制IEC-6细胞中的DNA结合活性和核NF-κB表达。总之，泼尼松龙和布地奈德在体外以剂量依赖性方式调节肠上皮细胞的修复机制，并深刻调节炎症调节因子NF-κB。
