废弃孜然根茎叶抗消化性溃疡的物质基础与作用机制研究

In China, The most of Cuminum cyminum are grown at Xinjiang, where has many large planting bases that account for more than 90% of total cumin planting area of this country. The Cuminum cyminum has been used in Uygur traditonal medicine for variaous diseases. Right now, the utilization of C. cyminum are focus on their fruits rather than other tissue (like root, stem and leaves). The C. cyminum other tissue is abandoned, which cause a huge waste of resources. Naphtha in other tissue (root, stem and leaves) of C. cyminum had been analysed for the first time in our previous study, the results showed that the main chemical composition of fruit also exists in the root, stem and leaves, in stem which the content of cuminaldehyde is almost twice as much as in fruit. The C. cyminum cuminaldehyde has been proved effective in therapy of several gastric ulcer models in rats by the mechanism of gastric mucosal recovery and protection, gastric acid reduction and proteolytic enzyme inhibition. The chemical constitutes and bioactivities of C. cyminum entire plant tissue (fruits, root, stem and leaves) had been investigated with pharmacology in this study to screen the lead compound and active constituent for the treatment of peptic ulcer and illustrate its possible mechanism and their interrelation. The objective of this study is to provide scientific basis for deep exploitation and clinical application of C. cyminum entire plant tissue (fruits, root, stem and leaves) and Uygur medicine, and meanwhile to promote the comprehensive application and wide development of C. cyminum and this potential medicine.

新疆是我国孜然的主要种植基地，占全国种植总面积的90％以上，资源非常丰富且又是新疆民族药。目前，孜然的利用主要集中在果实上，果实采摘后的根茎叶则废弃，不予重视，造成资源的极大浪费。前期研究中，我们首次对孜然根茎叶的挥发油进行了分析，结果发现孜然果实中主要的化学成分也存在于根茎叶中，其中对大鼠胃溃疡模型具明显的治疗作用的枯茗醛类物质，在根茎叶中含量均高于果实，而根中含量接近果实的2倍。因此，本项目拟对孜然全株（果实、根、茎、叶）展开系统的植物化学和生物活性研究，结合药效学研究，筛选出活性部位，寻找抗消化性胃溃疡活性成分，发现活性先导化合物，以期阐明其抗胃溃疡活性的物质基础、作用机制与构效关系，将为其在药物中的深度开发与临床应用提供重要的科学依据，同时可以为开发以孜然为主要成分的药品奠定一定的基础。希望能够以此来推动孜然的综合利用与发展。

本项目广泛地开展孜然药材的采集、鉴定及资源评价工作，开展孜然基因组测序和多样性分析，组装获得了孜然叶绿体和线粒体基因组序列，为孜然的遗传育种提供了基础资料；通过GC-MS对孜然果实、根和茎化学成分进行分析，共分离鉴定出30种成分，其中枯茗醛、2-Caren-10-al和4a,7,7,10a-Tetramethyldodecahydrobenzo[f] chromen-3-ol是孜然根茎最主要的抗菌、抗氧化和抗胃溃疡效应的重要活性成分；进一步比较评价孜然根茎不同的提取部位的药效活性差异，明确了孜然根茎抑菌活性主要集中在石油醚部位和乙酸乙酯部位，抗氧化活性主要集中在乙酸乙酯部位，抗胃溃疡的活性部位主要为乙酸乙酯部位和乙醇提取物部位，证实了孜然根茎化学成分和药效作用贡献的潜在效应物质；进一步，采用幽门结扎型大鼠胃溃疡模型，评价孜然根茎乙醇提取物抗胃溃疡作用效果与机制，结果发现根茎乙醇提取物对溃疡的有一定的保护作用，且能降低血清和胃组织SOD活性，抑制血清MDA生成，增加血清NO水平，增加血清GSH水平，同时升高胃组织NO来有效预防胃溃疡炎症的发生和发展；同时对孜然根茎中含量较高、具有潜在的活性成分进行提取纯化，通采用2种动物模型，考察了孜然中枯茗醛成分的对胃溃疡保护和机制，结果发现枯茗醛能够降低乙醇诱导的大鼠急性胃黏膜损伤模型的溃疡面积和胃黏膜损伤状况，增强机体抗氧化能力，抵抗炎症反应，同时加强胃黏膜防御的作用，达到抗胃溃疡的作用；同时，枯茗醛还能减缓吲哚美辛诱导的大鼠急性胃溃疡模型的炎症反应，具有较好的抗炎活性，其对胃黏膜保护作用与PGE2和NO有关；最后，通过高通量测序分析，构建长期灌胃枯茗醛对大鼠肠道微生物群落结构的相互作用网络，结果显示枯茗醛成分可以改善大鼠肠道菌群的结构，对肠道微生物的平衡发挥有益的影响，维持机体肠道微生物的稳态，对宿主的健康起积极作用。本项目对孜然果实和根茎中主要活性成分和活性部位有了较深入的体外和体内实验研究，系统比较研究了孜然根茎不同极性部位和活性成分对抗胃溃疡的作用效果和贡献程度，阐明了孜然根茎抗胃溃疡的药效学基础，包括有效的化学部位（乙酸乙酯部位和乙醇部位）和主要的单体化合物（枯茗醛），完成了孜然根茎的物质基础研究。本研究为揭示孜然根茎关键活性成分的挖掘与抗胃溃疡作用奠定了基础，为废弃孜然根茎资源化利用提供了新的方法。