The College of Pharmacy, Shaanxi University of Chinese Medicine, Xianyang, China.
The College of Life Sciences, Northwest University, Xi'an, China.
J Ethnopharmacol. 2024 Jan 10;318(Pt B):116961. doi: 10.1016/j.jep.2023.116961. Epub 2023 Aug 2.
Ilex rotunda Thunb. (IR) is widely used for gastrointestinal diseases by Yao physician, and it has a better clinical curative effect on ulcerative colitis (UC). However, the main active components and mechanism of IR in the treatment of UC remain to be clarified.
To investigate the main active components and mechanism of IR in the treatment of UC.
Ten biological active components of IR were quantified by UPLC-MS/MS. In vitro, Caco2 cell monolayers were stimulated by lipopolysaccharide, and were treated with 10 biologically active components individually to investigate the protective role of the components of IR in mucosal barrier damage. In vivo, a mouse model of UC was induced by dextran sulfate sodium and administered with the candidate active components of IR. On day 8, the serum and colon tissue were collected for histological and molecular analysis to investigate the main active components and mechanism of IR.
Ziyuglycoside I, ziyuglycoside II, syringin, and pedunculoside in IR reduced phenol red transmission of the monolayer, and inhibited the protein expression of oncostatin M and oncostatin M receptor in Caco2 cells. Notably, ziyuglycoside II and syringin decreased the transepithelial electrical resistance of the monolayer, and promoted the protein expression of Occludin, Claudin-1 and zonula occludens-1 (ZO-1) in Caco2 cells. In vivo, ziyuglycoside II and syringin improved the symptoms of UC mice, including body weight, disease activity score, shortening of colon length, damaging of acidic mucus layer, histopathological changes, and protein expression of Occludin, Claudin-1, and ZO-1. Pedunculoside reduced the neutrophils and inflammatory response in the UC mice. Moreover, when the combination of ziyuglycoside II, syringin and pedunculoside was used for the treatment of UC, syringin and pedunculoside enhanced the therapeutic effect of ziyuglycoside II. Finally, RNA sequencing and RT-qPCR analysis revealed that ziyuglycoside II + syringin + pedunculoside and IR coregulated up to 42.7% of genes, and mainly reduced the overexpression of C-X-C motif ligand 1(CXCL1), oncostatin M receptor (OSMR), interleukin 1 receptor type I (IL1R1), tumor necrosis factor receptor superfamily member 9 (TNFRSF9), C-X-C motif chemokine 13 (CXCL13), oncostatin M (OSM), and interleukin 6 (IL-6) in the cytokine-cytokine interaction pathways.
The combination of ziyuglycoside II, syringin, and pedunculoside protects against UC by modulating the intestinal mucosal barrier and inhibiting the cytokine-cytokine interaction pathways, and the effect is relatively equivalent to that of the water extract of Ilex rotunda Thunb.
苦丁茶（IR）被瑶医广泛用于治疗胃肠道疾病，对溃疡性结肠炎（UC）有较好的临床疗效。然而，IR 治疗 UC 的主要活性成分和机制仍有待阐明。
研究 IR 治疗 UC 的主要活性成分和机制。
采用 UPLC-MS/MS 对 IR 中的 10 种生物活性成分进行定量分析。在体外，用脂多糖刺激 Caco2 细胞单层，分别用 10 种生物活性成分处理，研究 IR 成分对黏膜屏障损伤的保护作用。在体内，用葡聚糖硫酸钠诱导 UC 小鼠模型，并给予 IR 的候选活性成分进行处理。第 8 天收集血清和结肠组织进行组织学和分子分析，以研究 IR 的主要活性成分和作用机制。
IR 中的梓醇苷 I、梓醇苷 II、丁香苷和潘托洛苷降低了单层中苯酚红的传递，抑制了 Caco2 细胞中肿瘤坏死因子-α（OSMR）和肿瘤坏死因子-α（OSM）受体的蛋白表达。值得注意的是，梓醇苷 II 和丁香苷降低了细胞单层的跨上皮电阻，促进了 Caco2 细胞中 Occludin、Claudin-1 和 zonula occludens-1（ZO-1）的蛋白表达。在体内，梓醇苷 II 和丁香苷改善了 UC 小鼠的症状，包括体重、疾病活动评分、结肠缩短、酸性黏液层损伤、组织病理学变化以及 Occludin、Claudin-1 和 ZO-1 的蛋白表达。潘托洛苷减少了 UC 小鼠的中性粒细胞和炎症反应。此外，当梓醇苷 II、丁香苷和潘托洛苷联合用于治疗 UC 时，丁香苷和潘托洛苷增强了梓醇苷 II 的治疗效果。最后，RNA 测序和 RT-qPCR 分析表明，梓醇苷 II+丁香苷+潘托洛苷和 IR 共同调节了多达 42.7%的基因，主要降低了细胞因子-细胞因子相互作用途径中细胞因子趋化因子配体 1（CXCL1）、肿瘤坏死因子-α（OSMR）受体（OSMR）、白细胞介素 1 受体 I（IL1R1）、肿瘤坏死因子受体超家族成员 9（TNFRSF9）、C-X-C 基序趋化因子 13（CXCL13）、肿瘤坏死因子-α（OSM）和白细胞介素 6（IL-6）的过度表达。
梓醇苷 II、丁香苷和潘托洛苷的组合通过调节肠道黏膜屏障和抑制细胞因子-细胞因子相互作用途径来预防 UC，其作用与苦丁茶水提取物相当。
