摘要：Purpose: To investigate the bioactive components and mechanism of action of moist exposed burn ointment (MEBO) for treatment of diabetic foot ulcers using network pharmacology (NP) and molecular docking technology Methods: Through pharmacology database of traditional Chinese medicine (TCM) systems (TCMSP), analysis platform and symptom mapping (SymMap) database, the bioactive components of MEBO were screened for and protein binding to bioactive components was predicted. Proteins related to Diabetic foot ulcer (DFU) were collected from GeneCards, OMIM, PharmGkb and TTD disease databases. With R language software, the binding proteins of bioactive components of MEBO intersected with the proteins related to occurrence of DFU. Proteins of DFU linked to treatment with MEBO were subjected to analysis using gene ontology (GO) and KEGG with R language software. AutoDock and PyMOL software were employed to dock active components of MEBO and major proteins of DFU. Targeted binding potential of bioactive compounds in MEBO to core proteins of DFU was analyzed. Results: One hundred and five (105) bioactive ingredients and 246 likely therapeutic proteins were obtained. Proteins were mainly involved in biological processes in wound healing, oxidative stress, and lipopolysaccharide. The enriched signaling pathway focused on lipid metabolism and the process of atherosclerosis which involved PI3K and Akt, advanced glycation end-products (AGE)-receptor (RAGE) and mitogen-activated protein kinase (MAPK). The absolute values of these proteins were screened out with a docking score greater than 5 kcal/mol. Conclusion: The biological effects of MEBO are related to multiple proteins and multiple signaling pathways. Therefore, healing effect of MEBO on DFU occurs via multiple pathways. The biological characteristics of MEBO need to be fully elucidated in further studies.

摘要：Background: The incidence of diabetes-related wounds is widespread, and the treatment is challenging. We found that Moist Exposed Burn Ointment (MEBO) promotes the healing of diabetes-related wounds, but the mechanism is not clear.Methods: This study aimed to explore the mechanism of MEBO on diabetic wound healing, which may be related to the promotion of re-epithelialization. A full-thickness skin resection model was established in streptozotocin(STZ)-induced diabetic mice. MEBO and Kangfuxin (KFX) were applied to the wound area, and the wound healing rate was analyzed by photographing. The granulation tissue and epidermal thickness, the collagen remodeling rate, and the expression of cytokeratin 10 (CK10), cytokeratin 14 (CK14), Ki67,CollagenⅠ,and Collagen Ⅲ in the regenerated skin were detected by H&E staining, Masson staining, and immunofluorescence staining, respectively. MEBO and KFX were applied to human immortalized keratinocytes (HaCaT)、mouse dermal fibrolasts (MDF) cells, and cell viability, cell migration, and differentiation were determined by CCK-8, scratching assay, RT-qPCR, and Western blot(WB), respectively. Results: We found that MEBO significantly promoted the formation of wound granulation tissue and collagen remodeling in diabetic mice. The application of MEBO to diabetic wounds not only promoted the formation of hair follicles and sebaceous glands but also promoted the expression of Ki67, CK10, and CK14 in epidermal cells. MEBO had no significant effect on the differentiation process of keratinocytes.Conclusions: Our study further proved that MEBO plays a positive role in diabetic wound healing, and its excellent ability to promote re-epithelialization may be an important reason for promoting wound healing.

摘要：目的:探讨湿润暴露疗法/湿润烧伤膏(Moist exposed burn therapy/moist exposed burn ointment,MEBT/MEBO)对糖尿病难愈合创面的作用,并以α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达水平反映糖尿病难愈合创面肌成纤维细胞转化程度,影响细胞外基质主要成分Ⅰ型胶原蛋白(Collagen I,COL-1)的生成为切入点,通过动物及细胞实验探讨MEBT/MEBO促进糖尿病难愈合创面的具体机制,为MEBT/MEBO的临床应用及推广提供理论依据。方法:(1)90只SPF级成年雄性Wistar大鼠以随机数表法分为五组,空白组、对照组、模型组、MEBO组、贝复新组,每组18只,适应性喂养7天后应用STZ腹腔注射法诱导糖尿病大鼠模型,成模后再以全层皮肤缺损法统一制备创面。观察创面愈合情况,拍照记录创面形成后第0、3、7、14天的大鼠创面,并随机切取每组6只大鼠创面肉芽组织,进行HE、Masson染色以观察各组创面组织病理形态变化,免疫荧光观察α-SMA、COL-1的分布及表达情况,Western blot检测α-SMA、COL-1的蛋白表达水平。(2)探讨MEBT/MEBO对大鼠真皮成纤维细胞(rat dermal fibroblasts,RDFs)的作用。首先用双酶法(中性蛋白酶+胰酶)分离提取Wistar大鼠幼崽皮肤真皮成纤维细胞,以HE染色观察细胞形态,免疫荧光法鉴定其波形蛋白(Vimentin)表达情况。以CCK8法摸索MEBO对RDFs的合适剂量,细胞实验分为低糖组、高糖组、溶剂组、低剂量MEBO组、高剂量MEBO组,DCFH-DA荧光探针法检测各组活性氧(reactive oxygen species,ROS)水平,实时定量PCR法检测α-SMA、COL-1的基因转录水平,Western blot法检测RDFs的α-SMA、COL-1蛋白表达水平。结果:(1)大鼠经STZ法诱导糖尿病模型后,出现典型“三多一少”症状,血糖明显升高达到糖尿病标准。与其他组相比,模型组大鼠出现创面愈合延迟(P<0.05),创面肉芽组织持续炎症浸润、血管新生不足、成纤维细胞数量低下及细胞外基质沉积减少等特征;治疗中后期创面愈合率:对照组(29)MEBO组=贝复新组(29)模型组。(2)WB:与对照组比较,创面发生第7、14天过程中,模型组创面组织α-...

摘要：糖尿病足溃疡（Diabetic foot ulcer DFU)是糖尿病患者晚期严重并发症之一，具有易诊断、难治疗、治疗周期长、医疗费用高且容易复发等特点，合并重度感染可导至患者残疾甚至死亡，给个人、家庭和社会造成了沉重负担。给予2021年4月28日南昌三三四医院普外科收治的1例糖尿病足溃疡合并重度感染患者，在全身综合治疗的基础上，局部全程用创疡再生医疗技术（MEBT/MEBO）治疗，先后两次住院，其余时间由医生指导家属在家换药，历时5个月，创面完全治愈，避免了截肢，保住了患者基本劳动能力。