HLA-G调控炎症反应促进组织工程血管长期通畅的研究

The immune response is closely related to angiogenesis, and a moderate inflammatory response can promote the rapid endothelialization of tissue engineered blood vessels (TEBV) and maintain its long-term patency. Human leukocyte antigen G (HLA-G) plays an important role in regulating the intensity and balance of inflammatory reaction. This project aims to construct a TEBV that can target endothelial progenitor cells (EPC) and deliver HLA-G protein or related microRNA. Cell experiments were used to study the effects of HLA-G on the mobilization, homing, proliferation, differentiation and paracrine of EPC. Based on previous studies, we constructed CD133 aptamer /HLA-G exosome, identified its ability of EPC capture and targeted HLA-G delivery, and further studied HLA-G regulating the interaction between EPC, regulatory T cells (Treg cells), monocytes / macrophages and smooth muscle cells and their mechanisms. Gold nanoparticles were synthesized and assembled in a layer by layer manner is used to construct CD133-PCS CD133 aptamer /HLA-G exosome K9 RNA chimeric modified TEBV scaffolds. TEBV was implanted into rats to study the effect of HLA-G on the long-term patency of TEBV, and to explore its mechanism, so as to provide theoretical support for understanding the interaction between cardiovascular system and immune system after TEBV implantation.

免疫反应与血管生成紧密相关，适度的炎症反应可促进组织工程血管（TEBV）快速内皮化和维持其长期通畅。人类白细胞抗原G（HLA-G）具有调控炎症反应强度与平衡的重要作用。本课题拟构建一种能靶向内皮祖细胞（EPC），并递送HLA-G蛋白或相关microRNA的TEBV。细胞实验研究HLA-G对EPC动员、归巢、增殖、分化和旁分泌的影响；基于前期研究，构建CD133适配体/HLA-G外泌体，明确其EPC捕获和靶向递送HLA-G的能力，并进一步研究HLA-G调控EPC、调节性T细胞（Treg细胞）、单核/巨噬细胞和平滑肌细胞之间相互作用及其机制；合成金纳米颗粒，层层组装的方式构建CD133适配体/HLA-G外泌体修饰的TEBV支架；TEBV植入大鼠体内，研究HLA-G对TEBV长期通畅的影响，并探讨其机制，为了解TEBV植入后心血管系统与免疫系统之间的相互作用提供理论支持。

免疫反应与血管生成紧密相关，适度的炎症反应可促进组织工程血管（TEBV）快速内皮化和维持其长期通畅。人类白细胞抗原G（HLA-G）具有调控炎症反应强度与平衡的重要作用。课题组成功分离和培养出（EPCs），并研究了HLA-G对EPCs、淋巴细胞和平滑肌细胞的作用；成功构建了适配体-ADK siRNA嵌合体/PEI/外泌体/HLA-G-miRNA纳米颗粒，体外和动物实验结果均显示其能够特异性结合靶细胞，并同时递送siRNA和HLA-G-miRNA；我们进一步将CD133适配体-ADK siRNA嵌合体/PEI/外泌体/HLA-G-miRNA纳米颗粒修饰到TEBV表面，动物实验结果显示其能通过调控炎症反应，促进糖尿病下TEBV的内皮化和长期通畅。基于碳纳米管和适配体技术，层层自组装的方式构建了一种能同时靶向递送siRNA和药物的胶原膜，动物实验结果显示其具有很好的治疗效果，这为后续构建特异性捕获EPCs、并靶向递送siRNA和HLA-G蛋白的抗疾病损伤TEBV提供了技术支撑。同时，我们探索了外泌体在抗高糖损伤的TEBV构建中应用，结果显示通过外泌体诱导神经长入和调控炎症反应可促进TEBV的长期通畅、以及在体重塑为成熟的血管。通过本课题，成功构建了一种CD133适配体-ADK siRNA嵌合体/PEI/外泌体/HLA-G-miRNA修饰的TEBV，并揭示了HLA-G通过调控炎症反应，促进TEBV长期通畅的作用及机制。这个项目为构建抗疾病损伤的TEBV提供了新的方法，为了解TEBV植入后心血管系统与免疫系统之间的相互作用提供理论支持。