目的 基于蛋白组学探讨冲和膏调控血管内皮生长因子（VEGF）通路促进糖尿病溃疡大鼠创面愈合的作 用机制。方法 采用高糖高脂饲料喂养+腹腔注射链脲佐菌素（STZ）+皮肤缺损的方法复制糖尿病溃疡大鼠模 型。将 SD大鼠随机分为空白组、模型组、冲和膏组、生长因子组，每组 6只。冲和膏组及生长因子组分别用 相应药物涂敷创面，每日给药1次，连续14 d。采用Masson染色法观察创面肉芽组织中胶原纤维生长情况；免 疫组化法检测创面肉芽组织中CD34、VEGF表达情况；通过蛋白组学方法检测模型组vs冲和膏组的差异表达蛋 白，并利用DAVID平台进行GO功能富集分析，利用KEGG数据库进行代谢通路富集分析；Western Blot法检测 创面肉芽组织中VEGF通路相关蛋白的表达水平。结果 与空白组比较，模型组大鼠创面肉芽组织的胶原纤维 分布稀疏，排列紊乱，胶原纤维蓝染较少，胶原纤维面积占比明显降低（P < 0.05）；CD34 标记的血管数目较 少，管径较细，微血管密度值及VEGF阳性表达显著下调（P < 0.05，P < 0.01）；Pxn、Casp9、Nos3、Ptk2蛋白 表达均显著上调（P < 0.01），VEGF蛋白表达显著下调（P < 0.01）。与模型组比较，冲和膏组及生长因子组大鼠 创面肉芽组织的蓝染胶原纤维沉积增多，排列整齐有序，胶原纤维面积占比显著升高（P < 0.01）；CD34标记的 血管数目较多，管径增粗，微血管密度值及 VEGF 阳性表达显著上调（P < 0.01）；Pxn、Casp9、Nos3、Ptk2蛋 白表达显著下调（P < 0.01），VEGF蛋白表达显著上调（P < 0.01）。与模型组比较，冲和膏组筛选出 221个差异 蛋白，其中28个上调蛋白，193个下调蛋白；主要涉及细胞骨架组织、超分子纤维组织、肌动蛋白细胞骨架组 织，以及VEGF、胰岛素抵抗、PI3K-Akt等信号通路。结论 冲和膏能够促进糖尿病溃疡大鼠的创面愈合，可 能与促进创面肉芽组织胶原纤维生成及血管新生，调控VEGF信号通路相关。

Objective To explore the mechanism of Chonghe Ointment in regulating vascular endothelial growth factor （VEGF）pathway to promote wound healing in diabetes ulcer rats based on proteomics. Methods A rat model of diabetes ulcer was established by the method of feeding high-sugar high-fat diet + intraperitoneal injection of streptozotocin（STZ）+ skin defect. SD rats were randomly divided into blank group，model group，Chonghe Ointment group and growth factor group， with six rats in each group. The rats in Chonghe Ointment group were applied with Chonghe Ointment once a day for successively 14 days. Masson staining was used to observe the growth of collagen fibers in the granulation tissues of wounds. Immunohistochemistry was used to detect the expression changes of CD34 and VEGF in the granulation tissues. Differentially expressed proteins between model group and Chonghe Ointment group were detected by proteomics. GO functional enrichment analysis was performed using the DAVID platform and metabolic pathway enrichment analysis were conducted using the KEGG database. Western Blot was used to detect the expression levels of VEGF-related protein in the granulation tissues of wounds. Results Compared with the blank group， the distribution of collagen fibers in the model group was sparse and disordered，less blue stained collagen fibers were seen and the proportion of collagen fiber area was significantly reduced（P < 0.05）.The number of CD34 labeled blood vessels was relatively small， the diameter was thin， and the microvascular density （MVD） value， and VEGF positive expression were significantly down-regulated（P < 0.05，P < 0.01）. The protein expression of Pxn，Casp9，Nos3 and Ptk2 were significantly up-regulated（P < 0.01）. The VEGF protein expression was significantly down-regulated（P < 0.01）. Compared with the model group，the collagen fibers in the Chonghe Ointment group and the growth factor group were arranged neatly and orderly，with more deposition of blue stained collagen fibers. The proportion of collagen fiber area was significantly increased（P < 0.01）. The number of CD34 labeled blood vessels was relatively large， the diameter was thick，MVD value and VEGF positive expression were significantly up-regulated （P < 0.01）. The protein expression of Pxn，Casp9，Nos3 and Ptk2 were significantly down-regulated（P < 0.01）. The VEGF protein expression was significantly up-regulated（P < 0.01）. Compared with the model group， a total of 221 differentially expressed proteins were screened in Chonghe Ointment group， including 28 up-regulated proteins and 193 down-regulated proteins， which mainly involve in multiple signaling pathways such as cytoskeleton tissue， supramolecular fibrous tissue， actin cytoskeleton tissue， VEGF， insulin resistance， PI3K-Akt， etc. Conclusion Chonghe Ointment can promote wound healing in diabetes ulcer rats. Its possible mechanism is related to promoting the formation and angiogenesis of collagen fibers in granulation tissues and regulating the VEGF signaling pathway.

R285.5

国家自然科学基金地区项目（82460899）；内蒙古自治区高等教育科学研究项目（NJZY23131）；内蒙古自治区草原英才团队 滚动支持项目（CYYCTD2024017）