School of Pharmacy, Key Laboratory of Molecular Pharmacology and Drug Evaluation (Yantai University), Ministry of Education, Collaborative Innovation Center of Advanced Drug Delivery System and Biotech Drugs in Universities of Shandong, Yantai University, 30 Qingquan Road of Laishan District, Yantai 264003, Shandong, PR China.
Department of Endocrinology, Yantaishan Hospital, Yantai 264000, PR China.
Phytomedicine. 2019 May;58:152864. doi: 10.1016/j.phymed.2019.152864. Epub 2019 Feb 18.
Ulcerative colitis (UC) is a chronic inflammatory condition of the intestines and is difficult to cure once diagnosed. The efficacy of the current clinical treatment for UC is limited. Common anti-inflammatory drugs are prone to adverse effects, while novel biological agents are expensive, although tolerated by patients. Therefore, an urgency exists to find more safe and effective drugs to treat UC. Osthole is an active constituent isolated from the fruit of Cnidium monnieri (L.) Cuss. Osthole has anti-inflammatory activities and offers certain intestinal protection. These characteristics indicate that osthole has the potential to inhibit UC.
The study was conducted to investigate the anti-inflammatory potential of osthole in LPS-induced RAW 264.7 cells and dextran sulphate sodium (DSS)-induced ulcerative colitis in mice.
In in vitro experiments, mouse monocyte-macrophage RAW 264.7 cells were stimulated by 1 μg/ml LPS to produce inflammatory mediators. Griess reagent was used to determine Nitric Oxide (NO) production, and ELISA kits were used to determine the levels of PGE TNF-α, and IL-6. The anti-inflammatory mechanisms of osthole were detected using western blot. In in vivo experiments, UC was induced via the intragastric administration of 3.5% DSS to BALB/C mice for 7 days. During the experiment, clinical signs and body weight were monitored and recorded daily to calculate the DAI score. At the end of the experiment, the colon lengths were measured. The colonic histopathological lesions were evaluated. MPO activity and TNF-α levels were determined using the corresponding kits. The protein expression of TNF-α and NF-κB pathways were analysed using western blot.
In an in vitro study, osthole inhibited the production of NO, PGE2, TNF-α, and IL-6 in LPS-induced RAW 264.7 cells. The results of western blot showed that osthole inhibited the expression of iNOS, COX-2, p38 MAPK and IκB α in RAW 264.7 cells. On this basis, in DSS-induced UC mice, it was found that osthole relieved the symptoms of UC by inhibiting weight loss, colon shortening and the DAI score, and simultaneously alleviating colon tissue lesions. It was also found that osthole reduced the levels of TNF-α in serum and colon tissues and effectively inhibited the activity of MPO. The western blot results showed that osthole reduced the expression of NF-κB p65 and p-IκB α and increased the content of IκB α in colon tissues.
Osthole exerted anti-inflammatory effects by blocking the activation of the NF-κB and MAPK/p38 pathways. Additionally, osthole possesses therapeutic potential in the treatment of UC.
溃疡性结肠炎（UC）是一种慢性肠道炎症性疾病，一旦确诊很难治愈。目前 UC 的临床治疗效果有限。常用的抗炎药容易产生不良反应，而新型生物制剂虽然患者耐受，但价格昂贵。因此，迫切需要寻找更安全有效的药物来治疗 UC。蛇床子素是从蛇床子（Cnidium monnieri（L.）Cuss）的果实中分离得到的一种活性成分。蛇床子素有抗炎作用，并能提供一定的肠道保护。这些特征表明蛇床子素具有抑制 UC 的潜力。
本研究旨在探讨蛇床子素在脂多糖（LPS）诱导的 RAW 264.7 细胞和葡聚糖硫酸钠（DSS）诱导的溃疡性结肠炎小鼠模型中的抗炎作用。
在体外实验中，用 1μg/ml LPS 刺激小鼠单核巨噬细胞 RAW 264.7 细胞产生炎症介质。用 Griess 试剂测定一氧化氮（NO）的产生，用 ELISA 试剂盒测定前列腺素 E2（PGE2）、肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的水平。用 Western blot 检测蛇床子素的抗炎机制。在体内实验中，通过灌胃 3.5%DSS 7 天诱导 UC 模型。实验过程中，每天监测和记录临床症状和体重，计算 DAI 评分。实验结束时，测量结肠长度。评估结肠组织病理学损伤。用相应试剂盒测定髓过氧化物酶（MPO）活性和 TNF-α 水平。用 Western blot 分析 TNF-α和 NF-κB 通路的蛋白表达。
在体外研究中，蛇床子素抑制 LPS 诱导的 RAW 264.7 细胞中 NO、PGE2、TNF-α 和 IL-6 的产生。Western blot 结果显示，蛇床子素抑制 RAW 264.7 细胞中 iNOS、COX-2、p38MAPK 和 IκBα的表达。在此基础上，在 DSS 诱导的 UC 小鼠中，发现蛇床子素通过抑制体重减轻、结肠缩短和 DAI 评分来缓解 UC 症状，同时减轻结肠组织损伤。还发现蛇床子素降低了血清和结肠组织中 TNF-α 的水平，并有效抑制了 MPO 的活性。Western blot 结果显示，蛇床子素降低了 NF-κB p65 和 p-IκBα的表达，增加了结肠组织中 IκBα的含量。
蛇床子素通过阻断 NF-κB 和 MAPK/p38 通路的激活发挥抗炎作用。此外，蛇床子素在溃疡性结肠炎的治疗中具有治疗潜力。
