Şen Ali, Özbeyli Dilek, Teralı Kerem, Göger Fatih, Yıldırım Aybeniz, Ertaş Büşra, Doğan Ahmet, Bitiş Leyla, Şener Göksel
Department of Pharmacognosy, Faculty of Pharmacy, Marmara University, Istanbul, Turkey.
Department of Medical Services and Techniques, Vocational School of Health Services, Marmara University, Istanbul, Turkey.
Chem Biol Interact. 2023 Jan 5;369:110263. doi: 10.1016/j.cbi.2022.110263. Epub 2022 Nov 12.
The aim of this study was to evaluate the therapeutic effect of active ethanol extract obtained from the leaves of Rubus tereticaulis (RTME) against colitis, and to purify major compounds from this extract by bioassay-directed isolation. Rats with colitis induced via intra-rectal acetic acid administration (5%, v/v) received RTME or sulfasalazine for three consecutive days. On day four, all rats were decapitated, and the colonic tissue samples were collected for macroscopic score, colon weight, reduced glutathione (GSH), myeloperoxidase (MPO), and malondialdehyde (MDA) analyses. The active compounds and chemical composition of RTME were determined by bio-guided isolation and LC-MS/MS, respectively. Compared to the colitis group, the rats treated with RTME displayed significantly lowered macroscopic scores and colon wet weights (p < 0.001). These effects were confirmed biochemically by a decrease in colonic MPO activity (p < 0.001), MDA levels (p < 0.001), and an increase in GSH levels (p < 0.001). Kaempferol-3-O-β-d-glucuronide (RT1) and quercetin-3-O-β-d-glucuronide (RT2) were found to be the major compounds of RTME, as evidenced by in vitro anti-inflammatory and antioxidant activity-guided isolation. Their anti-inflammatory/antioxidant activities were also predicted by docking simulations. Additionally, quinic acid, 5-caffeoylquinic acid, quercetin pentoside, quercetin glucoside, quercetin-3-O-β-d-glucuronide, kaempferol-3-O-β-d-glucuronide, and kaempferol rutinoside were identified in RTME via using LC-MS/MS. RT2, along with other compounds, may be responsible for the observed protective action of RTME against colitis. This study represents the first report on the beneficial effects of RTME in an experimental model of colitis and highlights the potential future use of RTME as a natural alternative to alleviate colitis.
本研究的目的是评估悬钩子叶活性乙醇提取物（RTME）对结肠炎的治疗效果，并通过生物活性导向分离法从该提取物中纯化主要化合物。通过直肠内注射乙酸（5%，v/v）诱导结肠炎的大鼠连续三天接受RTME或柳氮磺胺吡啶治疗。在第四天，所有大鼠断头处死，收集结肠组织样本进行宏观评分、结肠重量、还原型谷胱甘肽（GSH）、髓过氧化物酶（MPO）和丙二醛（MDA）分析。分别通过生物导向分离和液相色谱-串联质谱法（LC-MS/MS）测定RTME的活性化合物和化学成分。与结肠炎组相比，接受RTME治疗的大鼠宏观评分和结肠湿重显著降低（p < 0.001）。结肠MPO活性降低（p < 0.001）、MDA水平降低（p < 0.001）以及GSH水平升高（p < 0.001）从生化角度证实了这些效果。山奈酚-3-O-β-D-葡萄糖醛酸苷（RT1）和槲皮素-3-O-β-D-葡萄糖醛酸苷（RT2）被发现是RTME的主要化合物，体外抗炎和抗氧化活性导向分离证明了这一点。它们的抗炎/抗氧化活性也通过对接模拟进行了预测。此外，通过LC-MS/MS在RTME中鉴定出了奎尼酸、5-咖啡酰奎尼酸、槲皮素戊糖苷、槲皮素葡萄糖苷、槲皮素-3-O-β-D-葡萄糖醛酸苷、山奈酚-3-O-β-D-葡萄糖醛酸苷和山奈酚芸香糖苷。RT2与其他化合物一起，可能是RTME对结肠炎具有观察到的保护作用的原因。本研究是关于RTME在结肠炎实验模型中的有益作用的首次报告，并突出了RTME作为缓解结肠炎的天然替代品的潜在未来用途。
