壮药拔毒生肌膏激活特异性生长因子EGF、VEGF治疗慢性难愈创面的实验研究

Chronic refractory wound is a common chronic injury and regarded as a big problem in wound healing clinically. Wound healing process, which often effected by the infection includes necrosis tissue removable and wound regeneration. Wound regeneration is produced by the regulation of growth factor. Epidermal Growth Factor (EGF) plays a dominant role on the growth of epithelial cell, especially on the re-epithelialization in the middle and late stage of wound healing. Vascular endothelial growth Factor (VEGF), one of the key growth factors for the angiogenesis, is the strongest and only specific factor promoting angiogenesis by affecting vascular endothelial cells. To analyze the mechanism of the Budushengji cream in promoting Refractory Wound healing and to deeply and systematically study the mechanism of the traditional Chinese medicine in promoting wound healing. this research establishes chronic refractory wounds model on the diabetic rats ,oxidative stress rats and the rabbit ears which are the most popular diabetic and ischemic refractory wound model. the effect of the local classic prescription in Guangxi, Badushengji cream on EGF and VEGF of chronic refractory wound and the lysozyme content of wound exudative were investigated by ELISA, radioimmunoassay and immunohistochemistry. The quantity and morphology of fibroblast and new capillary in pathological image were observed as well and the effect of Badushengjigao cream on bacteria also be analyzed through AGAR medium ChuanDai culture method.

慢性难愈创面是最常见的慢性损伤之一，为临床一大难题。慢性难愈创面的愈合需经历坏死组织清除和组织再生修复两个进程。其进程全程受感染影响。组织再生修复受生长因子的调控。表皮细胞生长因子（EGF）对上皮细胞生长起主导作用，对创面愈合中后期的再上皮化有着特殊效果；血管内皮生长因子（VEGF）是影响血管新生的关键生长因子之一，是唯一特异性作用于血管内皮细胞的促进血管生成因子。本项目通过建立3种慢性难愈创面模型以模仿最为常见的糖尿病性、氧化应激性和缺血性难愈创面，应用ELISA、放射免疫法及免疫组化等先进技术，观察广西壮药拔毒生肌膏对模型创面EGF、VEGF及渗出液溶菌酶的影响；通过病理图文观察其对创面成纤维细胞与新生毛细血管的数量与形态的影响；通过琼脂培养基传代培养法观察其对细菌的影响，以探讨拔毒生肌膏治疗难愈创面的作用机制，对壮（中）医药促进难愈创面修复的机制进行深入系统研究。

慢性难愈创面是最常见的慢性损伤之一，为临床一大难题。壮药拔毒生肌膏能明显促进慢性难愈合创面生长。本项目通过建立3种慢性难愈创面模型以模仿最为常见的糖尿病性、氧化应激性和缺血性难愈创面，应用ELISA、放射免疫法及免疫组化等先进技术检测用药后大鼠及兔子血清溶菌酶、表皮生长因子（EGF）和血管内皮生长因子（VEGF）含量以及组织EGF和VEGF含量，通过病理图文观察组织病理变化，通过琼脂培养基传代培养法观察其对细菌的影响，明确拔毒生肌膏对糖尿病性难愈创面、氧化应激难愈创面和缺血性难愈创面 EGF、VEGF、创面肉芽组织病理结构的影响及对细菌的作用影响，初步阐明拔毒生肌膏治疗慢性难愈创面的分子作用机制及药物作用靶点。结果显示拔毒生肌膏能明显促进各种模型的创面愈合，结果基本符合实验设想，拔毒生肌膏通过激活EGF和VEGF促进创面愈合，病理组织显示实验组与模型组比较生长良好。本项目初步阐明拔毒生肌膏治疗慢性难愈创面与激活特异生长因子EGF和VEGF相关，我们现在正在进行实验的延展，使用免疫印迹（WB）检测技术和聚合酶链式反应（PCR）检测氧化应激模型大鼠创面新生肉芽组织PI3K和AKT表达情况，检测缺血性兔耳难愈创面新生肉芽组织TGF-β1和Smad3的表达情况。项目资助已发表论文两篇，待发表3篇。培养硕士生3名，其中一名已取得硕士学位，两名在读。项目投入经费48万元，支出45.851744万元，各项支出基本与预算经费相符。剩余经费2.148256万元，剩余经费计划用于本项目研究后续发表论文支出。