川芎-赤芍药对通过调控间充质干细胞向内皮分化影响血管新生及其机制研究

Although the replacement of new drugs and popularization of surgical operation have made more myocardial infarction(MI) patients benefit, some patients with MI still can not get effective treatment because of the diffuse lesions, postoperative restenosis and heart failure after MI. Stem cell transplantation has brought new methods for the treatment of ischemic heart disease. But there are many problems in the use of stem cells. One of the problems that needs to be solved is the low retention in situ and low-specificity of differentiation under pathological conditions. Previous work has proved that Rhizoma Chuanxiong, Red Peony and its effective components ligustrazine and paeoniflorin, could promote the angiogenesis of zebrafish, cerebral ischemia mice and endothelial cells effectively. Based on the previous work, the aim of this study is to preliminarily clarify the effect of couple of Rhizoma Chuanxiong and Red Peony on the angiogenesis of ischemic myocardium by regulating the differentiation of stem cells into endothelial cells. With the help of proteomics, molecular mechanism of Rhizoma Chuanxiong and Red Peony on angiogenesis will be deeply studied by regulating the differentiation of bone marrow mesenchymal stem cells into endothelial cells. This project will enrich the connotation of blood-activating and stasis-removing Chinese medicine promoting angiogenesis, and provide a new perspective and method for the application of stem cells in cardiovascular disease.

尽管新药的更替、外科手术的普及使更多的心梗病人获益，仍有部分患者因病变弥漫、术后再狭窄、心梗后心衰等原因，得不到有效治疗。干细胞移植为缺血性心脏病的治疗带来了新方法，但干细胞的应用还存在许多问题：原位保留率低、病理条件下分化的不专一性是待解决的问题之一。课题组前期工作已证实川芎、赤芍及其有效成分川芎嗪、芍药苷能有效促进基因缺陷斑马鱼、脑缺血小鼠及内皮细胞的血管生成。基于前期工作及川芎-赤芍药对在活血化瘀类方中的重要作用，本课题借助干细胞可向内皮细胞分化的特性，初步明确川芎-赤芍药对通过调控骨髓间充质干细胞，促进缺血心肌血管新生的作用。并借助蛋白质组学，部分揭示川芎-赤芍药对通过调控骨髓间充质干细胞向内皮分化，影响血管新生的分子机制，丰富活血化瘀中药促血管新生的内涵，为干细胞在心血管疾病中的应用提供全新的视角和方法。

治疗性血管新生是缺血性心脏病的有效方法。该项目在活血化瘀生新理论指导下，从川芎-赤芍药对促血管新生、修复心肌缺血损伤的现象着手，借助大鼠心梗及缺氧内皮细胞模型、蛋白组学技术，研究川芎-赤芍药对联合MSCs改善心梗大鼠心功能及促缺氧内皮细胞血管新生的药效机理。结果提示，川芎-赤芍药对联合MSCs较单用MSCs更好的改善心梗大鼠LVEF、FS及CO值，减轻心肌梗死面积，增加新生血管数量，提示川芎-赤芍药对与MSCs联用具有协同增效的作用；同时该药对含药血清促进缺氧内皮细胞的增殖、迁移和成管。蛋白组学结果显示，药对组与模型组比较，共得到154个差异表达蛋白，其中上调81个，下调73个。其中，CDC42/NRP1表达差异均较显著。CDC42参与包括angiogenesis、cell differentiation及sectretion过程，而NRP1/CDC42在内皮细胞迁移、出芽等过程中均发挥重要作用，提示CDC42介导的信号轴可能是川芎-赤芍药对介导内皮细胞与间充质干细胞交互作用，促进缺血心肌血管新生的关键，丰富了活血化瘀中药促血管新生的内涵，为后续研究提供了思路。GO富集分析结果显示“positive regulation of vasculature development”、“positive regulation of angiogenesis”差异显著，提示血管新生在改善心梗大鼠心功能的过程中发挥重要作用，与预期一致。此外，cell differentiation、secretion得分也较高，提示细胞分化、细胞分泌作用同时参与活血化瘀药对促血管新生的过程，但该分化和分泌作用是否与间充质干细胞有关，尚待深入。