桃红四物汤调节产后血瘀状态血小板alpha颗粒释放的物质基础及作用途径研究

Taohong-siwu decoction(TSD) has a definite therapeutic effect on Postpartum Blood Stasis Syndrome(PBSS), and the blood stasis has closely related to the abonormal activation and α-granule selectively release of platelets. Our previously research found that TSD could regulate release of α-granule in platelet, these may be related to TSD's therapeutic effect on PBSS. First, postpartum blood stasis rats model were establised by mifepristone and misoprostol induced abortion with blood index,the absorption ingredients into serum and the effect on α-granule release and active protein of platelet were analyzed. Second, the method of TSD platelet affinity extraction and HPLC-MS/MS analysis were established, and the active ingredients of TSD for regulating of platelet were determined. then the plasma sample of PBSS were collected and the character of α-granule release and active platelet protein were analyzed, then the different agonists were used to active platelet and the inhibitory effect of TSD extraction and affinity ingredients to actived platelet were analyzed, the acitived proteins in platelet stimulated by different agonists and the effect of TSD were determined by wstern blotting and flow cytometry instrument. Last, the active compound of TSD and its action pathway on platelet with PBSS were clarified. This research could interpret the mechanism of TSD on blood stasis syndrome from a new way.

桃红四物汤（TSD）在临床上对产后血瘀证具有明确的治疗作用，而血瘀证与血小板异常激活及α-颗粒选择性释放密切相关。前期研究发现TSD具有调节血小板释放α-颗粒作用，这可能是其治疗产后血瘀症的机制之一。本项目首先考察TSD醇提物对产后血瘀证大鼠血小板激活状态和α-颗粒分泌的影响以及TSD入血成分；然后建立TSD血小板亲和萃取HPLC-MS/MS分析方法，确定TSD作用于血小板的有效成分；同时采集临床产后血瘀证患者血液，分析血小板释放成分及激活状态；其次从血小板不同活化途径研究TSD有效成分对血小板聚集、α-颗粒分泌成分及血小板活化蛋白表达的影响；最终确定血瘀证状态下血小板α-颗粒分泌特征及TSD干预的物质基础及作用途径。本课题的研究有助于从一个新的角度全面分析TSD治疗血瘀证的机制。

血瘀证与血小板异常激活及α-颗粒选择性释放密切相关，本项目从一个全新的角度出发，分析产后血瘀状态下血小板α-颗粒释放蛋白特征，并分析桃红四物汤干预的物质基础及调控血小板作用途径。本项目研究分为六个部分：1）进行桃红四物汤质量控制研究。2）桃红四物汤干预血小板释放蛋白物质基础研究。3）桃红四物汤调控血小板释放作用途径研究。4）桃红四物汤调控血小板释放蛋白变化研究。5）血小板释放蛋白与子宫内膜修复关联性研究6）桃红四物汤对产后血瘀大鼠代谢组学的拓展性研究。本项目建立了可靠的桃红四物汤质量控制模式，鉴定桃红四物汤体外87个化学成分，发现与血小板结合5种成分；共鉴定出桃红四物汤治疗前与治疗后血小板差异蛋白39个，上调17个，下调22个；共鉴定出桃红四物汤治疗前与治疗后血小板释放差异蛋白92个，上调43个，下调49个；并通过动物实验证实血小板释放蛋白中IGF-1、SF-20和TSP-1、PF4在子宫中显著表达，促进子宫内膜修复。本研究证实桃红四物汤通过调节血小板α-颗粒释放蛋白，发挥抗血小板聚集、抗凝血、调节炎症反应的“祛瘀”作用，提高机体代谢功能、调节组织修复与血管新生的“生新”作用。已发表相关学术论文10篇，已接收但未发表文章1篇。其中包括SCI 5篇，核心期刊论文5篇；培养毕业博士生2名，毕业硕士生1名。