基于伤口微环境稳态的构建探讨化腐再生法促糖足创面愈合过程中筋之血化的机制

Diabetic foot infection often affects on the deep tissue of the tendon，the infection could spread along the tendon, ultimately the patients have to accept ampution and sometimes life-threatening. Therefore, the key of treatment is to deal with the tendon of diabetic foot ulcer correctly. Clinical observations showed that when dealing with the method of “Huafuzaisheng”, newborn visible granulation tissue was appeared on the surface of tendon which was degeneration but not completely lost activity, at last, the exposed tendon was completely covered by the granulation tissue, which was called “The Vascularization of Jin”. During this progress the wound healing eventually. We believe that the emergence of this phenomenon was established after the occurrence of wound microenvironment homeostasis of diabetic foot. This issue intends to make model using diabetic rats with tendon infection, study on micro nerve-immune-regulation of vascular wound microenvironment and make it reach the steady state.Through the integrated use of immunohistochemistry, RT-PCR, western blot, microvessel count and other technical. Through the dynamic changes of VEGF and its receptors, the detection of TGF- beta, bFGF, BMP, HoxD, MVC, nerve growth factor NGF and its receptor Trk A, substance SP, M1, M2 type of macrophage related cytokines and other inflammatory factors, the aim of which was to reveal the mechanism of “The Vascularization of Jin” using the method of “Huafuzaisheng” in order to further promote the more sugar foot wound to find entry points.

糖尿病足破溃经常深达肌腱，且感染会沿肌腱蔓延，患者被迫截肢甚至失去生命。因此糖尿病足创面中肌腱的外科处理是治疗成功与否的关键。临床发现，经化腐再生法治疗后，创面中变性但尚未失去活性的肌腱表层上可见散在分布的新生肉芽岛，并逐渐生长，最终暴露的肌腱被新生的肉芽组织覆盖，称之为“筋之血化”，经过此过程后创面愈合。我们认为筋之血化的出现是建立在糖尿病足创面微环境稳态形成之后。本课题建立糖尿病大鼠伤口伴肌腱感染模型，研究神经-免疫-血管调节伤口微环境，并使其达到稳态，通过综合运用免疫组化技术、RT-PCR、western blot、微血管计数等技术，检测VEGF及其受体、TGF-β、bFGF、BMP、HoxD、MVC、神经生长因子NGF及其高亲和力受体TrkA、SP物质、M1、M2型巨噬细胞相关因子及其他炎性因子的动态变化，揭示化腐再生法促进糖尿病足筋之血化的机制，为促愈糖足创面寻找新的切入点。

糖尿病足是糖尿病严重并发症之一，创面往往造成足部肌腱的暴露，并随着感染的加重发生变性乃至坏死，使得创面在短时间内进一步扩大，为了挽救生命，患者被迫截肢甚至失去生命。如何控制变性坏死肌腱的进一步恶化，降低致残率及致死率，降低患者及社会的经济负担，是目前国内外医学界亟待解决的重要问题。临床发现，应用化腐再生散靶向去除坏死肌腱及组织，结合生肌象皮膏，给予创面湿性愈合环境，可以加速创面生肌长皮，促进肉芽组织、皮肤组织原位再生。经化腐再生治疗后，创面中变性但尚未失去活性的肌腱表层上可见散在分布的新生肉芽岛，并逐渐生长，并以络脉长入的方式使其与肌腱、筋膜连为一体，进而包埋滋养之，继而暴露的肌腱被新生的肉芽组长覆盖，直至与血自里到外融为一体，称之为“筋之血化”，最终使得创面愈合。通过实践发现认为筋之血化现象的出现是建立在糖尿病足创面微环境稳态形成之后。故研究化腐再生法调控创面微环境的构建与筋之血化出现的关系，对于提高糖尿病足创面修复水平具有重要意义。在此研究背景下查阅相关文献梳理了糖尿病伤口筋之血化现象出现与微环境稳态的建立及创面“神经调节--免疫调节-微血管生成”（神经-免疫-血管系统）的关系和微环境改善局部的症状以及中医药干预治疗糖尿病足的方法和原理的研究进展现况等方面的相关文献，探讨中医基础理论对糖足微环境的认识，并进行糖尿病大鼠溃疡创面微环境稳态的构建与筋之血化过程中不同时间点神经-血管-免疫相关因子的动态变化规律研究，通过运用病理学、分子生物学、免疫组织化学等技术和方法，形态学与功能性研究相结合，整理并分析实验结果，研究创面微环境稳态构建的微观机制，探讨糖尿病伴肌腱坏死创面筋之血化时局部创面微环境稳态构建的物质基础，从血管生成、免疫反应、神经调节等方面综合评价，为中医“筋之血化”理论提供客观依据，丰富糖尿病足创面愈合的结点和机制，为对于完善和发展中医疮疡的治疗具有重要意义。