功能化自组装肽缓释药物和优化干细胞移植对梗死心肌修复的促进作用及其机制

The myocardial infarction threatens people's life and health. However, there is no desirable therapy for the disease. It is very important for treatment of myocardial infarction how to promote exogenous myocardium repairing and activate endogenous cardiac regeneration. Viewing that stem cells are difficult to survive after transplantation and mechanism of endogenous repairing is not clear, this program is to develop a new functioned self-assembling peptide and prepare a system releasing of Tβ4、RA or RA and BMP-2 and compound of carrying stem cells for transplantation,and to explore possibility of activating endogenous cardiac regeneration and promoting exogenous repairing. In experiments in vitro, the biological characteristics of the functioned self-assembling peptide and releasing of the drugs and cytokine are evaluated, the effects of the released drugs and cytokine on proliferation and differentiation of stem cells, epicardium-derived cells and cardiomyocytes are investigated. After animal models of the myocardial infarction are set up, effectiveness of the released drugs and cytokine and the carried stem cells on promoting exogenous myocardium repairing and activating endogenous cardiac regeneration is evaluated in vivo. This program is to focus on exploring mechanism of activating regeneration of the epicardium and myocardium with the functioned self-assembling peptide releasing drugs. The program is valuable in regeneration medicine and clinical application for exploring a way of activating endogenous myocardium repairing, formulating strategies of optimizing stem cell transplantation, elucidating mechanism of exogenous and endogenous myocardium repairing and improving significantly therapeutic efficacy of myocardial infarction.

心肌梗死严重威胁着人们的生命与健康，但目前尚无理想治疗措施。如何有效促进外源性和激活内源性心肌修复对于治疗心肌梗死至关重要。鉴于干细胞移植后难以存活和分化而内源性修复机制还不清楚，本项目拟研制新型功能化自组装肽，制备Tβ4、RA等药物缓释系统和承载干细胞的移植复合物，探索促进外源性心肌修复同时激活心脏内源性再生的可行性。体外评价功能化自组装肽的生物学特性和缓释作用，检测缓释的药物和细胞因子对干细胞、心外膜源性细胞和心肌细胞增殖和分化等的作用。建立心肌梗死动物模型，在体评价功能化自组装肽缓释药物和细胞因子以及承载干细胞移植促进外源性和激活内源性心肌再生的效果。重点探讨功能化自组装肽缓释的药物激活心外膜和心肌内源性修复的机制。本项目对于探索内源性心肌修复的激活途径、制定干细胞移植优化策略、阐明外源性和内源性心肌修复的机制和显著提高心肌梗死的治疗效果具有重要的再生医学意义和临床应用价值。

本项目按照原计划要求，圆满完成了各项原计划研究内容，达到了预期指标。完成计划概要：①成功地设计与合成了结合药物Tβ4、RA和细胞因子BMP-2的功能化自组装肽，通过分子模拟、高效液相色谱和质谱检测等综合鉴定，确保所合成的功能化自组装肽的分子质量符合实验要求。②利用原子力显微镜和扫描电镜观察了自组装肽的自组装性能和超微结构特征；采用高效液相色谱法和ELISA法检测了药物和细胞因子从自组装肽的缓释，综合评价了功能化自组装肽的生物学特性；通过动物皮下注射和免疫学检测评价了功能化自组装肽的组织相容性、免疫原性和降解过程等，建立了可靠的纳米生物材料特性评价指标，为后续利用自组装肽缓释药物促进梗死心肌修复提供了可行性依据。③通过建立体外细胞培养模型评价了药物缓释对心外膜源性细胞（EPDCs）和MSCs增殖和分化以及对心肌细胞（CMs）分裂和增殖的作用，为体内通过Tβ4或RA缓释激活心外膜和成年CMs分裂以促进内源性心肌再生奠定了可靠基础。④通过建立了大鼠心肌梗死模型，在体评价了药物缓释和优化干细胞移植对移植细胞存活和分化的促进作用及其机制，为临床采用干细胞移植治疗心肌梗死提供了可靠的实验依据。⑤ 通过建立了大鼠和转基因小鼠心肌梗死模型，探讨了通过Tβ4或RA缓释激活心外膜和启动成年CMs分裂促进内源性心肌修复的可行性。这对于有效利用内源性心肌修复机制修复梗死心肌提供可行性依据具有重要意义。本项目研究成果已发表SCI收录论文9篇，其中SCI期刊封面文章1篇，已完成并投稿SCI论文3篇。发表中文核心期刊论文7篇。国际学术会议交流论文8篇，国内学术会议交流论文6篇。获科技成果奖1项（上海市科学技术奖自然科学奖三等奖），申请专利2项（申请中）。在国际学术会议作专题和分组报告4次。在全国性学术会议作专题报告1次，分组报告5次。