茭白壳提取物对小鼠溃疡性结肠炎的预防作用

本研究通过乙醇浸提、硅胶色谱层析得到茭白壳提取物（Zizania latifolia shell extract，Zlse），探究其对小鼠溃疡性结肠炎的影响，并通过网络药理学对Zlse预防小鼠溃疡性结肠炎的机制进行初步分析。实验设置正常组、模型组、阳性对照组和Zlse低、中、高剂量处理组（Zlse-L组、Zlse-M组、Zlse-H组），模型组、阳性对照组、Zlse组通过灌胃3.5%（质量分数）葡聚糖硫酸钠溶液建立溃疡性结肠炎模型，阳性对照组和Zlse组分别进行美莎拉嗪和Zlse灌胃处理，正常组和模型组给予0.5%（质量分数）羧甲基纤维素钠溶液灌胃，每天记录小鼠健康情况，进行疾病活动指数（disease active index，DAI）评分，比较6 组小鼠体质量变化率、DAI评分、结肠长度、苏木精-伊红染色切片、组织病理学评分、炎症因子（白细胞介素（interleukin，IL）-6、IL-8、IL-1β、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α））水平、髓过氧化物酶（myeloperoxidase，MPO）活力、超氧化物歧化酶（superoxide dismutase，SOD）活力；通过GeneCards、OMIM、TTD和DAVID数据库筛选关键靶点及重要京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）通路，构建“活性成分-靶点-KEGG通路”网络图。结果表明，相较于模型组，Zlse组体质量降低趋势显著减缓（P＜0.05），各组结肠长度有所恢复，结肠损伤减轻，此外，Zlse组血清中各促炎症因子水平显著降低（P＜0.05），结肠组织MPO活力显著降低（P＜0.05），SOD活力显著升高（P＜0.05）；通过网络药理学分析获得8 个活性成分、77 个交集靶点和119 条KEGG通路，关键活性成分为小麦黄素及小麦黄素-4’-O-丁香醇醚，其中重要通路包括磷脂酰肌醇3-激酶/苏氨酸激酶信号通路、受体激活和活性氧积累等，与动物实验结果相印证。结果表明Zlse能够通过抑制促炎症因子的释放，调控结肠抗氧化防御能力，预防葡聚糖硫酸钠盐诱导的小鼠溃疡性结肠炎，且其功效与浓度呈正相关。

Preventive Effect of Zizania latifolia Shell Extract on Ulcerative Colitis in Mice

In this study, Zizania latifolia shell extract (Zlse) was obtained by ethanol extraction and silica gel chromatography, and its effect on ulcerative colitis in mice was evaluated. The underlying mechanism was analyzed by network pharmacology. Six groups of mice were established: normal control (NC), model control (MC), positive control (PC), low-dose Zlse (Zlse-L), medium-dose Zlse (Zlse-M) and high-dose Zlse (Zlse-H). Ulcerative colitis was induced by intragastric administration of 3.5% sodium dextran sulfate solution in the MC, PC and Zlse groups. The PC and Zlse groups were intragastrically administrated with mesalazine and Zlse, respectively. The NC and MC groups were intragastrically administrated with 0.5% sodium carboxymethylcellulose (CMC-Na) solution. The health status of mice was recorded daily, and the disease activity index (DAI) score was calculated. The percentage body mass change, DAI score, colon length, hematoxylin and eosin (HE)-stained colonic sections, colonic histopathology score, serum inflammatory factors (IL-6, IL-8, IL-1β, and TNF-α) levels, and the activities of colonic myeloperoxidase (MPO) and superoxide dismutase (SOD) were compared between the six groups. Based on the key targets and the important Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways obtained from the GeneCards, OMIM, TTD and DAVID databases, a ‘active ingredient-target-KEGG pathway’ network diagram was constructed. The results showed that compared with the MC group, the trend of body mass loss in the Zlse treatment groups was significantly slowed down (P < 0.05), the colon length was restored, and the colon injury was reduced. In addition, the levels of serum proinflammatory factors in the Zlse groups were significantly decreased (P < 0.05); colonic MPO activity was significantly decreased (P < 0.05), while SOD activity was increased (P < 0.05). Eight active components, 77 intersection targets and 119 KEGG pathways were obtained by network pharmacology analysis. The key active components were tricin and tricin-4’-O-syringyl alcohol, and the important pathways included the P13K/Akt signaling pathway, receptor activation and reactive oxygen species accumulation, which were confirmed by animal experiments. The results showed that Zlse can concentration-dependently prevent DSS-induced ulcerative colitis in mice by inhibiting the release of proinflammatory factors and regulating the antioxidant defense capacity of the colon.