基于“创面内源性蛋白酶水解显效”作用特征的珍珠促进糖尿病创面愈合活性多肽（群）的发现研究

Wound healing injury caused by diabetes mellitus is one of its complications and a major challenge in clinical treatment. In the past decades, with the continuous development of various new drugs, new technologies and new treatment strategies, the treatment effect of diabetic wound healing has been improved to some extent, but the long treatment cycle and low short-term wound healing rate can not be fundamentally solved. Therefore, it is of great significance to explore new treatment drugs and methods for the prevention and treatment of diabetic wounds. Previous studies have shown that Pearl polypeptide is the main effective component of pearl in accelerating wound healing of diabetes mellitus. The effect of Pearl polypeptide is indirectly exerted by the hydrolysis of protease contained in wound exudate, which produces polypeptide with smaller molecular weight. Based on this feature, this project intends to explore the active polypeptides (groups) of pearls that promote diabetic wound healing by using the strategy of "component knockout/knockin " on the basis of previous studies. At the same time, proteomics and bioinformatics methods are used to analyze the types of endogenous proteases and their active polypeptides on the wound. The enzymatic hydrolysis of Pearl polypeptide reveals the unique role of Pearl polypeptide in promoting diabetic wound healing, and provides preliminary data support for the subsequent development of new small molecular active polypeptide drugs to promote diabetic wound healing.

糖尿病引起的创面愈合损伤是其并发症之一，也是临床治疗的一大挑战。在过去的数十年间，随着各种新药物、新技术和新治疗策略的不断发展，糖尿病创面不愈的治疗效果获得一定改善，但治疗周期长，短期创面愈合率低等问题任然无法得到根本性解决，因此，探寻新的治疗药物和治疗方法对于糖尿病创面防治具有重要的意义。课题组前期研究表明，珍珠多肽为珍珠加速糖尿病创面愈合的主要起效成分，其起效是经创面所含蛋白酶水解作用后，产生分子量更小的多肽间接发挥作用。基于此作用特征，本项目拟在前期研究的基础上，采用“成分敲出/敲入”策略，以糖尿病大鼠全皮层切除动物为模型，探寻珍珠发挥促进糖尿病创面愈合的活性多肽（群）；同时，采用蛋白质组学和生物信息学的方法分析创面内源性蛋白酶种类及其对珍珠活性多肽的酶解作用，揭示珍珠多肽促进糖尿病创面愈合的独特作用过程，并为后续开发出新的促进糖尿病创面愈合的小分子活性多肽药物提供前期数据支持。

糖尿病引起的创面愈合损伤是其并发症之一，也是临床治疗的一大挑战。在过去的数十年间，随着各种新药物、新技术和新治疗策略的不断发展，糖尿病创面不愈的治疗效果获得一定改善，但治疗周期长，短期创面愈合率低等问题任然无法得到根本性解决，因此，探寻新的治疗药物和治疗方法对于糖尿病创面防治具有重要的意义。课题组前期研究表明，珍珠多肽为珍珠加速糖尿病创面愈合的主要起效成分，其起效是经创面所含蛋白酶水解作用后，产生分子量更小的多肽间接发挥作用。基于此作用特征，本项目首次预测到糖尿病创面内源性蛋白酶Collagenase type 3 、Elastase-2可能酶解珍珠蛋白Nacrein-like protein F （NLF），产生小分子活性多肽Na-1。之后，本研究采用真核表达系统对Na-1进行了异源表达，动物实验结果表明，Na-1重组多肽有显著促进糖尿病大鼠全皮层切除创面愈合的作用。上述研究揭示珍珠多肽促进糖尿病创面愈合的独特作用过程，并为后续开发出新的促进糖尿病创面愈合的小分子活性多肽药物提供前期数据支持。