基于“心开窍于舌”探索硝石雄黄散调控口腔菌介导的硝酸盐还原酶保护缺血心肌的研究

The “tongue is the orifice of heart” is one of the important theory in Traditional Chinese Medicine. It plays an important role to prevent the cardiovascular disease .Xiaoshixionghuang powder is a typical representative of using this theory .Although it has a remarkable clinical effect but its mechanism is still unclear. Based on the previous relevant investigations and the advance knowledge in cardiovascular, We integrate the theory of “tongue is the orifice of heart” with oral bacterial and latest research results in nitric oxide (NO) research together. We proposed the hypothesis that Xiaoshixionghuang powder protects the ischemic myocardium by regulating the oral nitrate level, the oral anaerobia and its relative nitrate reductase activity to produce robust NO bioactivity through Nitrate-Nitrite-NO pathway. In order to confirm this hypothesis ,Our team develop this study from molecular level , protein , cell , organ and whole body by making use of 16S DNA sequencing, HPLC, immunoblotting, ultrasound and pathological tissue detection methods. Once the hypothesis is confirmed, This study will reveal the nature of the theory “tongue is the orifice of heart” as well as the mechanism of Xiaoshixionghuang powder how to protect ischemic myocardium from a new view of the relationship among the oral bacterial 、Nitrate-Nitrite-NO pathway and CAD. This research will also provide a new strategy and new ideal for prevention and treatment of CAD. This study will provide a theoretical and experimental basis for further studying and improving the Xiaoshixionghuang powder.

“心开窍于舌”是传统中医的重要理论，在心血管疾病的防治中起着重要的作用。治疗急心痛（冠心病）的古方硝石雄黄散正是运用这一理论的典型代表，疗效显著但机理未明。课题组基于前期研究和冠心病领域的前缘知识，将“心开窍于舌”理论与口腔细菌、一氧化氮研究的最新成果融合在一起，提出“硝石雄黄散籍借调控口腔中硝酸盐的含量、口腔厌氧菌及其介导的硝酸盐还原酶的活性，通过硝酸盐—亚硝酸盐—一氧化氮通道产生强劲的一氧化氮生物活性，保护缺血心肌”的假说。利用16SDNA测序、高效液相、免疫印迹、动物超声以及病理检查等技术，从分子、细胞、蛋白、组织和整体动物等多层面递进式的展开研究。一旦假设被证实，将会从口腔细菌及其介导的硝酸盐还原酶的活性与CAD的相关性的全新角度揭示“心开窍于舌”的科学内涵和硝石雄黄散的作用机制，为冠心病的防治提供一条崭新的研究思路和策略，为进一步研发和提升硝石雄黄散提供理论依据和实验基础。

“心开窍于舌”是传统中医的重要理论，在心血管疾病的防治中起着重要的作用。治疗急心痛（冠心病）的古方硝石雄黄散正是运用这一理论的典型代表，疗效显著但机理未明。课题组基于前期研究和冠心病领域的前缘知识，将“心开窍于舌”理论与口腔细菌、一氧化氮研究的最新成果融合在一起，提出“硝石雄黄散籍借调控口腔中硝酸盐的含量、口腔厌氧菌及其介导的硝酸盐还原酶的活性，通过NO3ˉ-NO2ˉ-NO通道产生强劲的NO生物活性，保护缺血心肌”的假说。本研究以硝石雄黄散散为研究药物，利用分子生物学、药物分析等手段从分子、蛋白、细胞、组织及整体动物水平多层次递进式的展开研究。药物分析结果表明：硝石原本为硝酸钠/钾，雄黄本身含有少量硝酸盐。当小鼠在含服硝石雄黄散后，口腔内的NRs酶活性明显增强，且小鼠的唾液、胃液和尿液中NO3-/NO2-的含量明显增加；细胞实验显示：含硝石雄黄的血清干预后的缺氧受损细胞内NO3-/NO2-/NO含量升高最显著；离体主动脉环血管反应性实验显示：硝石雄黄散具有明显的血管舒张作用，其中最高舒张比值达到100%；在心肌缺血小鼠模型中发现，模型小鼠经硝石雄黄散干预后心功能明显好转，病理组织检测结果显示有明显保护心肌细胞凋亡的作用。本研究最终验证了当机体出现缺氧状态时，“硝石雄黄散调控口腔菌群促进硝酸盐还原，通过NO3ˉ-NO2ˉ-NO通道产生强劲的一氧化氮生物活性保护缺血心肌”的假说。为冠心病的防治提供了一条新的研究思路和策略，为开发具有中医特色的原创新药、提升传统方剂疗效以及助推中药走向国际市场提供理论依据和实验基础。