基于肺泡巨噬细胞网状调节的祖卡木颗粒组方配伍及作用靶点预测

The clinical value of Uighur Medicine is very important, but the experimental design of Uighur medicine prescription is relatively inadequate at present, and the research on its compatibility mechanism through objective indicators is also very scarce. Based on the previous work, this project introduces the theory of "Monarch Feudatory Ambassador " in Traditional Chinese Medicine and takes Zukamu Keli as an example drug to establish the research hypothesis of " Alveolar macrophages reticulocyte regulation ". It is proposed to carry out the work from two levels. On the one hand, compared with the phagocytosis-secrete-immune effect of Alveolar macrophages, the drug and the adjuvant drugs, the aim and the difference of the AM phagocytosis-secretory-immune were analyzed. By analyzing the moving tendency of Alveolar macrophages, the change of content, the ultrastructure. Observe MCP-1, MIP-1α, ET-1, Ang-2, VEGF, detection of Alveolar macrophages phagocytosis of erythrocytes and natural killer cells, analysis of T lymphocyte proliferation and transformation levels, to deduce the principle and compatibility of Zukamu Keli mechanism. On the other hand, using metabolomics and cellular immunology as research techniques, the key targets of Zukamu Keli are predicted from both the whole lung fibrosis and local two-dimensional alveolar macrophages. Through this study, following the experience of Uighur Medicine experts, using Traditional Chinese Medicine language to describe the combination of Weiwei drugs, integrating cell identification technology to predict the target, multi-disciplinary integration, and new perspectives, it is conducive to the construction of new methods for medical research.

维吾尔医药临床价值非常重要，但目前应用科学方法阐释维药组方的实验设计相对不足，通过客观指标表征配伍机理的研究比较匮乏。本项目在前期工作基础上，以祖卡木颗粒为示例药物，将中医“君臣佐使”理论融入维医药研究，建立“肺泡巨噬细胞网状调节”工作假说；体内实验比较君药、臣药、佐使药三组药物对AM“吞噬-分泌-免疫”递进式网状调节的差异，分析AM移动趋向、含量变化、超微结构、内膜流动性，观察MCP-1、MIP-1α、ET-1、Ang-2、VEGF，检测鸡红细胞和自然杀伤细胞、观察T淋巴细胞增殖转化，根据三组药对“网状调节”的特异性选择作用，推演祖卡木颗粒的组方原则和配伍机理；体外实验，比较含药血清对AM、IM形态增殖分泌影响，分析祖卡木颗粒对细胞作用机制。本研究充分尊重维医药专家经验，尝试用中医语言表述维药组方配伍，整合细胞辨识技术预测作用靶点，多学科融合、新角度诠释，有利于构建维医药研究新方法。

祖卡木颗粒具有独特的临床疗效，但目前应用科学方法阐释其药效机制的实验设计相对不足。本项目围绕肺泡巨噬细胞（AM）“吞噬-分泌-免疫”网状调节这一科学假说，推演祖卡木颗粒的组方原则和配伍机理，分析祖卡木颗粒的抗炎与免疫调节机制，主要科技创新体现在以下三方面：.第一，创新构建中药（民族药）复方“药性分析、质量控制、机理阐释和理论创新”四位一体的研究新模式。维吾尔药大多性热、无毒、有矫正药和代用药等，借鉴中医学“七情和合”“君臣佐使”理论，祖卡木颗粒既针对病因、病症使用疏散风热、清热解毒、止咳平喘的药物，同时又使用大枣、山奈以养护脾胃，后续研究应以实现“药性临床广泛认可，质控体系不断健全，作用机理深入阐释，理论应用创新发展”为重点。.第二，创新整合一种基于细胞因子平衡的双向免疫调节网络模块复合分析的研究新方法。以ALI、IPF为病理模型，祖卡木颗粒通过双向免疫调节发挥免疫稳态调节和器官保护的作用，急性损伤阶段使过度分泌的Th1细胞因子向Th2偏移，纤维化阶段使过度分泌的Th2细胞因子向Th1偏移。祖卡木颗粒能有效抑制ALI大鼠肺组织CD14和共刺激分子CD80、CD86 mRNA的表达，抑制机体过激的免疫炎症反应。.第三，创新提出一种通过调节AM“自噬-凋亡”的平衡治疗特发性肺纤维化的研究新策略。以IPF为病理模型，祖卡木颗粒能够抑制肺泡巨噬细胞增殖、降低其凋亡；干预Bcl-2与Beclin 1的结合、抑制Caspase 3的活化；刺激吞噬功能的增强，抑制TLR4/MyD88/NF-κB表面受体信号通路的高表达，调节肺泡炎期间AM“自噬-凋亡”的平衡，从而在一定程度上延缓了纤维化过程，验证了祖卡木颗粒对AM具有递进式“网状调节”作用的研究假说。.综上所述，本项目成果有自主知识产权，研究模式具有示范性，获中华中医药学会科技进步奖三等奖1项，发表高质量学术论文7篇（其中4篇被SCI收录），研制并发布中华中医药学会团体标准10项，培养科研助理1人，毕业硕士研究生2人，促进了维药药性理论研究团队的可持续发展。