生肤油促进Ⅲ度烫伤皮肤无瘢痕修复的机制研究

Deep thermal injuries result in the loss of a physical barrier to pain, infection, progressive necrosis and scar formation. However, there is no a better understanding of the molecular mechanisms behind these processes may provide insight into developing improved wound healing therapies. ShengFu Oil is a Chinese herbal compound that is widely used to treat various burns and scalds. Our previous studies suggest that SFO could stimulate the skin's regeneration and help to reduce excess scars and to enhance scarless repair. It is therefore urgent to research the mechanism of the skin regeneration and scarless repair by SFO. In this project, we will determine the expressions of β-catenin and substance P in the skin tissues of full-thickness burn model during wound repair using some techniques including radioimmounassay, image analysis and RT-PCR, etc. Furthermore, we will investigate the mRNA and protein expression levels of those factors described above and analyze the relationship between β-catenin and substance P, and investigate influences of those factors on the cell proliferation of rat cultured hair follicle stem cells and the apoptosis inhibition of fibroblasts derived from human pathological scars. The results will conclude the co-regulatory effects of β-catenin and substance P on the skin regeneration and wound scar healing. Next, we will investigate the regulatory effects of SFO on the signal transduction between β-catenin and substance P, and to identify SFO able to accelerate "Shengji" and "Quyu" in the skin tissues of full-thickness burn wounds. The results described above will clarify molecular mechanism of scarless wound healing enhanced by SFO. The results of this project contributes to a better understanding of skin wound healing theory, provide new targets for the development of drugs for the treatment of burn wounds.

深度烧烫伤常导致创面皮肤剧痛、坏死、感染、瘢痕等难症，目前缺乏高效治疗药物，且相关药物的治疗机制尚未明了，反过来阻碍高效新药的开发。为此，本项目针对Ⅲ度烫伤皮肤具有明显治疗功效的中药复方生肤油，研究其促进深度烫伤皮肤无瘢痕修复的分子机制。分别建立Ⅲ度烫伤动物模型、毛囊干细胞和成纤维细胞的不同培养系以及抗感染检测体系，利用免疫组化、核酸定量等技术，检测烧烫伤后创面及周边皮肤组织中以及各培养体系中β连环素、P物质等蛋白、核酸的表达量，研究其对毛囊干细胞增殖与分化、成纤维细胞凋亡抑制等过程的调控关系，揭示β连环素、P物质共同调控创面早期再生及后期瘢痕修复的基本规律，并研究生肤油影响β连环素与P物质间信号传导并促进皮肤"生肌、祛瘀"的规律，阐明生肤油促进Ⅲ度烫伤创面皮肤无瘢痕修复的分子机制。本项目的研究结果有助于更好地从理论上认识创伤皮肤的修复过程，为烧烫伤治疗新药的开发提供新的靶点。

深度烧烫伤常导致创面皮肤剧痛、坏死、感染、瘢痕等难症，目前缺乏高效治疗药物，且相关药物的治疗机制尚未明了，反过来阻碍高效新药的开发。为此，本项目针对Ⅲ度烫伤皮肤具有明显治疗功效的中药复方生肤油，通过建立动物和细胞水平的实验模型，利用免疫组化、RT-PCR等成熟技术，检测生肤油对伤后各期创面皮肤组织中β连环素、COX-2等表达的调控规律，从β连环素与COX-2间信号传导的角度，研究该系统调控皮肤再生、瘢痕修复的分子机理，阐明生肤油促进Ⅲ度烧烫伤皮肤无瘢痕修复的分子机制。.依据所定研究目标，经过项目团队成员四年的努力，我们检证了生肤油可通过在创面修复期先刺激β-catenin、Dlk1蛋白表达，抑制炎症因子COX-2蛋白表达，在创面修复中期抑制β-catenin蛋白持续高表达，调控Dlk1和COX-2蛋白的表达，在创面修复末期维持β-catenin“活化”，即通过调控“β-catenin与COX-2”的炎性环路信号的异常活化，从而促进皮下成纤维细胞增殖、重塑，加快创面皮肤附属器的再生，抑制病理性瘢痕产生，达到促进皮肤创面无瘢痕修复的良好疗效。在生肤油的抗炎镇痛作用与机制研究中，基于通过成纤维细胞划痕试验验证结果，我们发现生肤油的组分之一乳香的油提取物能促进细胞迁移，而且通过小鼠的耳廓肿胀、足底热痛阈的实验检测，观察到乳香的油提取物有良好的抗炎镇痛效果。再基于乳香提取物中有效成分的GC-MS分析与抗炎镇痛试验的对应结果分析，发现乳香正是通过其所含的α-蒎烯、芳樟醇、正辛醇等有效成分抗炎镇痛，促进细胞迁移而发挥重要作用，是生肤油不可缺少的重要组分之一。另外，我们也观察到生肤油联合降糖药或生肤油单独局部给药均能够较好地改善糖尿病性末梢循环障碍引起的皮温下降以及痛觉迟钝，并通过上调VEGF表达，促进患部皮肤的血管重建的同时，下调COX-2表达，降低创面皮肤炎性反应，促进糖尿病足溃疡创面愈合。综合我们研究结果，生肤油基于其良好的抗炎、镇痛以及抗菌的作用，干预“β-catenin与COX-2”的炎性环路信号，从而达到了促进创面皮肤快速、无痕地修复。目前生肤油的相关技术已经转化了，希望通过市场推广能为更多人减轻痛苦。