基于肠道菌群与血小板活性探讨星蒌承气汤干预脑梗塞急性期痰热腑实证的效应机制研究

Patients of acute stroke with the syndrome of phlegm heat and bowel excess (PHBE) are those that could have constipation, which influence conscious state and nervous functional defects. Xing-Lou-Cheng-Qi decoction presented by professor Wang Yongyan is effective for stroke with PHBE. Stool property could be influenced by stroke with PHBE, and intestinal flora gut microbiota and platelet activity maybe found in patient of stroke with PHBE. However, it is not clear that how gut microbiota and platelet function have effect on the stroke with PHBE. New researches found that TMAO, metabolite of gut microbiota could enhance platelet activity and accelerate thrombogenesis and risk of stroke. So in this research, we suppose that the machnism of Xing-Lou-Cheng-Qi decoction treating stroke of PHBE would be related to adjust intestinal flora gut microbiota and platelet activity. We will use 16SrDNA sequencing and thromboelastography to evaluate gut microbiota and platelet activity. Compare the difference of gut microbiota and platelet activity between stroke patients with and without constipation, and before and after the different treatment, to verify the hypothesis above. Therefore, for the mechanism of acute stroke of PHBE and the efficacy of xing-lou-cheng-qi decoction based on gut microbiota and platelet function, this study may provide some objective data and new thought, which may have significance for treatment and prognosis of stroke.

脑梗塞急性期痰热腑实证（以下简称“腑实证”）即中风伴有大便秘结或不通，对患者意识状态及神经功能缺损程度有较大影响，王永炎院士提出的星蒌承气汤治疗腑实证具有明确疗效。腑实证引起大便性状改变，可能出现肠道菌群紊乱与血小板功能亢进，而星蒌承气汤治疗腑实证在肠道菌群与血小板活性层面的效应机制尚不明确。研究发现肠道菌群代谢产物氧化三甲胺（TMAO）能间接提高血小板活性，进而促进血栓形成，增加脑卒中发生风险。故本研究认为腑实证形成以及星蒌承气汤治疗腑实证可能与调节肠道菌群和血小板活性有关。应用16SrDNA测序技术和血栓弹力图等检查，对比分析腑实证和中风非痰热腑实证模型大鼠、比较应用星蒌承气汤和常规通便法干预腑实证前后及两组间肠道菌群分布及血小板活性的差异，以验证上述假说。从而为星蒌承气汤治疗腑实证在肠道菌群及血小板活性层面的效应机制提供客观数据及新思路，对脑卒中的治疗和预后有重要意义。

本研究旨在通过观察肠道菌群和血小板活性的变化，探讨星蒌承气汤治疗急性脑梗死痰热腑实证的疗效机制，为痰热腑实证的形成和星蒌承气汤的效应机制提供数据支撑。将大鼠分为A组（空白组）、B组（中风非痰热腑实证组）、C组（痰热腑实证组）、D组（星蒌承气汤组）、E组（乳果糖组）、F组（阿司匹林组）和G组（氯吡格雷组），对每组采集血样和粪便分别进行血栓弹力图检测和16s rRNA测序，以观察血小板活性和肠道菌群的变化。共分为三个主要研究内容，其结果如下：一、痰热腑实证和星蒌承气汤对肠道菌群的影响，结果表明痰热腑实证组大鼠菌群多样性显著降低，某些致病菌属丰度增加，而星蒌承气汤能恢复、改善肠道菌群的多样性，提高某些有益菌群的丰度，这可能是星蒌承气汤达到“通腑”效果的机制；二、痰热腑实证和星蒌承气汤对血小板活性的影响：痰热腑实证组血小板活性增加，血小板AA抑制率最小，处于高凝状态，而星蒌承气汤组能显著降低血小板活性，增加血小板AA/ADP抑制率，起到较好的抗血小板作用，这可能是星蒌承气汤改善中风症状的机制；三、中风病痰热腑实证和星蒌承气汤干预后血小板与肠道菌群的相关性分析：痰热腑实证组随着对血小板抑制程度减弱，则菌群覆盖程度减少，而星蒌承气汤组R值延长，相当于使用了“抗凝剂”，进而菌群多样性增加，故星蒌承气汤肠道菌群多样性的变化与血小板活性呈现正相关，且能通过调节痰热腑实证紊乱的菌群丰度，加强抗血小板作用。本研究研究结论认为痰热腑实证的形成与肠道菌群多样性降低、血小板活性增高有关，而星蒌承气汤能够增加肠道菌群多样性、降低血小板活性，增加血小板抑制率，这可能是星蒌承气汤治疗痰热腑实证起到化痰通腑作用的效应机制，为其在调节肠道菌群和抗血小板层面提供了证据支撑，也为进一步探讨肠道菌群代谢产物相关机制奠定基础，对脑卒中的治疗和预后有重要意义。本项目已基本按照课题计划书完成，研究过程中培养主治医师2名，目前发表核心期刊论文1篇。