Abstract：
Methotrexate (MTX)-induced gastrointestinal mucositis is a common adverse effect characterized by redox imbalance and overproduction of inflammatory mediators that perturb intestinal integrity. Currently, there is no definitive treatment for this condition and its prevention is still far beyond comprehension. Because of its pleiotropic pharmacological actions, we aimed to explore the potential mechanisms through which cilostazol (CILO) can protect against MTX-induced intestinal mucositis. Wistar rats were allocated into 4 groups, control, CILO (100 mg/kg, p.o for 14 days), MTX (7.5 mg/kg for 4 successive days), and CILO + MTX. The improving effect of CILO on the morphological structure was confirmed by an upturn in the histopathological and transition electron microscope examinations evidenced by the increased jejunal villus height/width and the crypt depth besides the maintenance of tight junctions. These findings were verified biochemically; on the molecular level, CILO reduced the MTX-induced lipid peroxidation, cleaved caspase-3, p53, and the inflammatory parameters (TLR-2, NF-κB, IL-23, TNF-α, IL-1β), while increasing the anti-inflammatory marker IL-10 and the antioxidant enzyme SOD. Moreover, CILO decreased the injurious axis AKT/GSK-3β/cyclin-D1, and CD44+, but increased the immunoexpression of the cell proliferating marker PCNA. CILO also upheld the intestinal barrier by enhancing the tight junction molecules (ZO-1, claudin-4) and the E-cadherin/β-catenin complex while abating the mesenchymal marker vimentin. In conclusion, CILO protected gut integrity by reducing the epithelial-mesenchymal transition process, the MTX-induced oxidative, apoptotic, and inflammatory mediators, and turning off the CD44/AKT/GSK-3β/cyclin D1 trajectory and intensifying the expression of PCNA.
摘要：
甲氨蝶呤（MTX）引起的胃肠道粘膜炎是一种常见的不良反应，其特征是氧化还原失衡和过度产生的炎症介质会扰乱肠道完整性。目前,这种情况没有明确的治疗方法，其预防仍然远远无法理解。由于其多效的药理作用，我们旨在探讨西洛他唑(CILO)预防MTX诱导的肠黏膜炎的潜在机制.Wistar大鼠分为4组,control,CILO（100mg/kg，p.ofor14days),MTX（连续4天7.5mg/kg），和CILO+MTX。组织病理学和过渡电子显微镜检查的上升证实了CILO对形态结构的改善作用，除了维持紧密连接外，空肠绒毛高度/宽度和隐窝深度也增加了。这些发现得到了生化验证；在分子水平上，CILO降低了MTX诱导的脂质过氧化，裂解的caspase-3，p53和炎症参数（TLR-2，NF-κB，IL-23,TNF-α,IL-1β)，同时增加抗炎标志物IL-10和抗氧化酶SOD。此外，CILO降低损伤轴AKT/GSK-3β/细胞周期蛋白-D1和CD44+，但增加了细胞增殖标志物PCNA的免疫表达。CILO还通过增强紧密连接分子（ZO-1，claudin-4）和E-cadherin/β-catenin复合物来维持肠屏障，同时减少间充质标记物波形蛋白。总之,CILO通过减少上皮-间质转化过程保护肠道完整性，MTX诱导的氧化，凋亡，和炎症介质，关闭CD44/AKT/GSK-3β/cyclinD1轨迹，增强PCNA的表达。
