人烧伤皮肤胞外基质活性肽的炎症免疫诱导作用及其对创面愈合和汗腺再生影响的实验研究

Degradation products of extracellular matrix (ECM) have strong inflammatory immune inductive activity. Whereas, the mechanisms of how the bioactive peptides in the burned skin ECM(bs-ECM) take part in wound healing and sweat glands regeneration are still obscure. So on the basis of our previous research, we will take normal skin (NS) of adult human being as controls, and the following researches will be carried out: (1) Extract the pooled protein of human burned skin in vitro, screen and identify the efficient inflammatory immune transmitters of human bs-ECM, and the chemotaxis and induction activities to human peripheral blood mononuclear cells (PBMCs) and bone marrow mesenchymal stem cells (BMSCs) will be assessed for some important inflammatory immune mediators among them. (2) The models of human thick split-thickness wound with removal of the epidermis and superficial dermis and retain of sweat glands，and deep Ⅱ degree burn of nude mice will be duplicated. On this basis, the chemotaxis and induction effects of inflammatory immune mediators and their neutralizing antibodies by local intervention on PBMCs and BMSCs which are labeled with 5-ethynyl-2′-deoxyuridine (EdU) will be assessed, respectively. Through the methodologies such as protein analysing, molecular pathology, immune-tracking and and other techniques, we will comparatively study the induced effects of these factors on the the recruitment and differentiation-inducing and activating functions of nonhomologous bone marrow-derived cells in different wounds, as well as how it effecting on the phenotype, secretion and structural feature of heterogeneous epithelial cells and fibroblasts. Ultimately, we can discuss the immune-inflammatory mechanism of sweat glands microenvironment (mainly ECM) alteration and pathological wound repairing and tissue regeneration, and provide a new theoretical and practical basis for perfect wound repairing.

细胞外基质(ECM)降解物具有强烈的炎症免疫诱导活性。然而，人烧伤皮肤ECM(bs-ECM) 活性肽在其创面愈合和汗腺再生中的作用机制不清。在前期工作的基础上，以成人正常皮肤为对照，⑴离体烧伤皮肤混合提取液中炎症免疫活性肽的筛查和鉴定，并就其中几个重要的炎症免疫介质进行人外周血单个核细胞(PBMCs)和骨髓间充质干细胞(BMSCs)趋化诱导实验；⑵建立保留汗腺人皮肤中厚断层创面和深Ⅱ度烫伤创面的裸鼠模型，并分别进行主要炎症免疫介质、中和抗体对5-乙炔基-2'脱氧尿嘧啶核苷标记的人PBMCs、BMSCs趋化诱导的局部干预实验。通过蛋白分析、分子病理和免疫示踪等技术，对比研究这些因子对骨髓源性细胞募集、分化和功能活化的介导效应，及其对创面异质性上皮细胞和成纤维细胞表型、分泌和结构特征的影响，阐明以ECM为主的汗腺微环境改变和病理性创面修复的炎症免疫机制，为创面完美修复提供新的理论和实践依据。

细胞外基质(ECM)降解物具有强烈的炎症免疫诱导活性。然而，人烧伤皮肤ECM(bs-ECM) 活性肽在创面愈合的免疫诱导和汗腺再生中的作用机制不清。本课题组通过蛋白分析、分子病理和免疫示踪等技术，对比研究这些因子对骨髓源性细胞募集、分化和功能活化的介导效应，探索了以ECM为主的汗腺微环境改变和病理性创面修复的炎症免疫机制，为创面修复提供新的思路。经过3年的探索研究，本课题组已成功构建多种动物模型，包括成人皮肤裸鼠移植模型、小鼠异体皮移植模型、胎儿皮肤裸鼠移植模型，为后续实验的开展奠定基础；成功提取并标记小鼠BM-MSCs，经细胞因子预处理后探索其对同种异体皮瓣移植的作用；探索并优化了DADM的获取方法，拓展DADM在烧伤治疗中的应用前景；成功提取出胎儿真皮间充质干细胞（FD-MSCs）并发现其对瘢痕疙瘩成纤维细胞的抑制作用；筛选噬菌体模拟肽调控局部创面的愈合等。.以上成果说明了以皮肤ECM为主的汗腺微环境改变对损伤修复和组织再生的炎症免疫调节机制，为创面完美修复提供新的理论、思路和实践依据，为进一步的科研计划奠定坚实的基础。