从Src/VE-cadherin通路研究清利法调控湿热型创面修复的作用机制

Incision in anal and rectal operation is contaminated wound.It has significantly inflammatory exudation ,and the healing of the wounds was more difficult.Inflammatory response in wound after operation often because of damp-heat,in spite of which symptom it is before surgery.That red swelling,heat pain and exudation of wound is he symptom of damp-heat, and vascular permeability increased is the key of its modern pathogenesis.The degree of permeability relate to both inflammatory exudation and the wax and wane of damp-heat.The study of open wound around anus must pay attention to inflammatory and exudation.Most study of wound healing view to tissue repair at home and abroad,and there was less study on influence of exudation on wound healing.Based on Summary of the experiments of professor Bai Liansong who is famous TCM doctor in china, the studying team study on therapeutic mechanism of external washing method for clearing heat and eliminate dampness.They Primarily found that regulate vascular permeability is important link of Improve the syndrome of inflammatory exudation , damp-heat and promoting wound repair.Research of Foreign also display vascular permeability is important link of regulate inflammatory exudation and promoting wound repair,it is a important field. We from optical microscope ,laser confocal,mRNA and protein level angle to study the mechanism that external washing method for clearing heat and eliminate dampness regulate vascular permeability promoting inflammatory exudation though Src/VE-cadherin signal pathway.And explore the mechanism that Traditional Chinese Medicine maybe regulate vascular permeability control inflammatory to promote wound healing,and so as to find more Scientific and Effective external washing method of TCM.

痔科手术切口是开放污染伤口，炎症渗出明显，不易愈合，是临床难题。不论术前是何症候，术后伤口局部炎症表现中医多责之为湿热。我们前期研究表明：伤口红肿热痛渗出的湿热证象，其现代医学的核心改变之一是血管通透性增加。通透性改变既和炎症渗出有关，又和局部湿热之邪盛衰与愈合密切关联。研究痔科开放伤口愈合应更重视炎症和渗出。国内外伤口愈合研究大多着眼组织修复，关注通透性对愈合影响的甚少。课题组总结全国名中医柏连松教授经验，研究清热利湿外洗疗效机制，初步发现调节血管通透性是改善局部炎症渗出和湿热证象，促进愈合的重要环节。国外研究也显示血管通透性在调节炎症渗出、促进愈合两方面都起重要作用，值得重视。本项目从中药影响Src/VE-cadherin通路调控血管内皮紧密连接机制入手，从光镜、电镜、mRNA、蛋白及磷酸化表达等水平研究清利外洗改善伤口炎症，控制湿热的机制，为进一步提高临床疗效和新药研发提供基础。

痔科手术切口是开放污染伤口，炎症渗出明显，不易愈合。不论术前是何症候，术后伤口局部炎症多责之以湿热。伤口红肿热痛渗出的湿热证象，其现代病机的核心改变是 血管通透性增加。通透性程度既和炎症渗出相关，又和局部湿热之邪盛衰密切相关。研究肛旁开放伤口愈合，更应重视炎症和渗出。国内外伤口愈合研究大多着眼组织修复，就炎症渗出对愈合影响研究较少。课题组在总结全国名中医柏连松教授经验基础上，研究清热利湿外 洗疗效机制，初步发现调节血管通透性是改善局部炎症渗出和湿热证象，促进愈合的重要环 节。本项目从光镜、激光共聚焦、mRNA、蛋白表达等水平研究清利法外洗通过 Src/VE-cadherin 信号通路调控血管渗透性以改善局部炎症（湿热）的机制，探讨中医药从 调控血管渗透性控制炎症，促进愈合的可能机制，以期找到更科学有效的中医药外治方法。.研究以SD大鼠肛旁创面模型为研究对象，在术后第3、7、11天通过伊文氏蓝染色法观察创面毛细血管通透性情况；肉眼及电镜观察创面渗出、炎症及愈合情况；电镜下观察血管内皮细胞紧密连接及胶原纤维合成、分布情况。结果提示造模后第3天清热利湿组与虎杖苷组伊文氏蓝渗出情况无差异（P>0.05），但两者渗出均少于生理盐水组（P<0.05）；第7，11天三组伊文氏蓝渗出差异无统计学意义（P>0.05）。肉眼观察发现造模后第3，7，11天三组创面分泌物渗出差异无统计学意义（P>0.0167）。术后第3、7天清热利湿组肉芽增生及上皮爬行优于生理盐水组（P<0.0167），但与虎杖苷组无差异（P>0.0167）；虎杖苷组与生理盐水组差异均无统计学意义（P>0.0167）。术后第11天三组创面愈合情况差异均无统计学意义（P >0.0167）。光镜结果可见清热利湿方组肉芽组织无明显水肿，新生毛细血管丰富且成纤维细胞排列整齐，虎杖苷组与之接近，两者愈合情况均优于生理盐水组。电镜结果可见清热利湿方组血管内皮细胞紧密连接将相邻细胞紧密嵌合；胶原纤维分布致密、规则，纤维束排列紧密。Elisa实验结果表明清热利湿法能够有效地降低血管通透性，减少创面组织 水肿，控制创面炎性渗出，促进肉芽组织胶原纤维的合成，提高创面愈合率；同 时能够有效促进创面组织中 VE-cadherin 的基因和蛋白表达，这可能是清热利湿 药外洗控制炎症，改善局部湿热症状，促进愈合的分子基因机制。