METHODS: The DSS-induced colitis mouse model was established by administration of 2.5% DSS for 5 days. Mice were orally administered with IA-0130 (0.01 mg/kg or 0.1 mg/kg) or cyclosporin A (CsA; 30 mg/kg). Body weight, disease activity index score and colon length of mice were calculated and histological features of mouse colonic tissues were analyzed using hematoxylin and eosin staining. The expression of inflammatory cytokines and tight junction (TJ) proteins were analyzed using quantitative real-time PCR and enzyme-linked immunosorbent assay. The expression of interleukin-6 (IL-6) signaling molecules in colonic tissues were investigated using Western blotting and immunohistochemistry (IHC).
RESULTS: IA-0130 (0.1 mg/kg) and CsA (30 mg/kg) prevented colitis symptom, including weight loss, bleeding, colon shortening, and expression of pro-inflammatory cytokines in colon tissues. IA-0130 treatment regulated the mouse intestinal barrier permeability and inhibited abnormal TJ protein expression. IA-0130 down-regulated IL-6 expression and prevented the phosphorylation of signaling molecules in colonic tissues.
CONCLUSIONS: This study demonstrated that IA-0130 suppressed colitis progression by inhibiting the gp130 signaling pathway and expression of pro-inflammatory cytokines, and maintaining TJ integrity.
方法:通过给予2.5%DSS5天建立DSS诱导的结肠炎小鼠模型。小鼠口服给予IA-0130(0.01mg/kg或0.1mg/kg)或环孢菌素A(CsA;30mg/kg)。体重,计算小鼠的疾病活动指数评分和结肠长度,并使用苏木精和伊红染色分析小鼠结肠组织的组织学特征。使用定量实时PCR和酶联免疫吸附试验分析炎性细胞因子和紧密连接(TJ)蛋白的表达。使用蛋白质印迹和免疫组织化学(IHC)研究结肠组织中白细胞介素-6(IL-6)信号分子的表达。
结果:IA-0130(0.1mg/kg)和CsA(30mg/kg)可预防结肠炎症状,包括减肥,出血,结肠缩短,和促炎细胞因子在结肠组织中的表达。IA-0130处理调节小鼠肠屏障通透性并抑制TJ蛋白异常表达。IA-0130下调IL-6表达并阻止结肠组织中信号分子的磷酸化。
结论:本研究表明IA-0130通过抑制gp130信号通路和促炎细胞因子的表达来抑制结肠炎的进展,保持TJ的完整性。