Department of Cell Biology, College of Basic Medical Sciences, Dalian Medical University, Dalian, China.
Department of Orthopedic Surgery, Second Affiliated Hospital, Dalian Medical University, Dalian, China.
PLoS One. 2024 Sep 6;19(9):e0309903. doi: 10.1371/journal.pone.0309903. eCollection 2024.
Diabetic skin wound is a complex problem due to the disruption of normal repairing program and lack of effective remedy. Lucilia sericata larvae (maggot) is a folk method to treat chronic skin wound, while its therapeutic effects on that caused by diabetic remains unknown.
This study aims to investigate the therapeutic effects of maggot extract (M.E.) on diabetic skin wound and its molecular mechanism by establishing the skin wound model of diabetic Sprague Dawley (SD) rats.
Diabetic model was established by injecting intraperitoneally streptozotocin in SD rats under specific pathogen-free (SPF) conditions. The rat fasting blood glucose values were ≧16.7 mmol/L 72 hours after intraperitoneal streptozotocin (60mg/kg body weight) injection. The rats were divided into five groups (n = 10/group): normal group: normal SD rats without any treatment, diabetic blank group: the diabetic rats without any treatment, Vaseline group: the diabetic rats dressed with Vaseline, recombinant human epidermal-growth-factor (rhEGF) group: the diabetic rats dressed with a mixture of Vaseline and 200 μg/g rhEGF, M.E. group: the diabetic rats dressed with a mixture of Vaseline and 150 μg/ml maggot extract. The round open wounds (1.0 cm in diameter) down to the muscle fascia were made on both sides of rat dorsa by removing the skin layer (epidermis and dermis) and were daily photographed for calculating their healing rates. Hematoxylin-eosin (HE) and Masson's trichrome staining were performed on skin wound sections to analyze re-epithelialization and granulation tissue formation. Immunohistochemical (IHC), immunofluorescent (IF) stainings and Western blotting were conducted to analyze the statuses of STAT3 signaling.
The average wound healing rates on the 14th day were 91.7% in the normal, 79.6% in M.E., 71% in rhEGF, 55.1% in vaseline and 43.3% in the diabetes blank group. Morphological staining showed more active granulation tissue formation, re-epithelialization and neovascularization in M.E.-group than those in the blank and the vaseline-treated groups. Decreased p-STAT3 nuclear tranlocation and down-regulated Bcl-2, CyclinD1 and vascular endothelial growth factor (VEGF) expression were evidenced in the diabetic rats, which could be improved by rhEGF and especially M.E.
Maggot extract would be an alternative and/or adjuvant candidate for the better management of diabetic skin wounds because of its activity in enhancing STAT3 activation.
糖尿病皮肤创面是一个复杂的问题,由于正常修复程序的中断和缺乏有效的治疗方法。丝光绿蝇幼虫(蛆)是一种民间方法来治疗慢性皮肤创面,而其对糖尿病引起的创面的治疗效果尚不清楚。
本研究旨在通过建立糖尿病 SD 大鼠皮肤创面模型,探讨蛆提取物(M.E.)对糖尿病皮肤创面的治疗作用及其分子机制。
在特定病原体(SPF)条件下,通过腹腔注射链脲佐菌素(STZ)建立糖尿病模型。腹腔注射 STZ(60mg/kg 体重)72 小时后,SD 大鼠空腹血糖值≥16.7mmol/L。将大鼠分为五组(n=10/组):正常组:无任何处理的正常 SD 大鼠;糖尿病空白组:无任何处理的糖尿病大鼠;凡士林组:糖尿病大鼠涂凡士林;rhEGF 组:糖尿病大鼠涂凡士林和 200μg/g rhEGF 混合物;M.E.组:糖尿病大鼠涂凡士林和 150μg/ml 蛆提取物混合物。通过去除皮肤层(表皮和真皮)在大鼠背部两侧制作直径为 1.0cm 的圆形开放创面,并每天拍摄照片以计算其愈合率。对皮肤创面切片进行苏木精-伊红(HE)和 Masson 三色染色,分析再上皮化和肉芽组织形成情况。采用免疫组织化学(IHC)、免疫荧光(IF)染色和 Western blot 分析 STAT3 信号状态。
第 14 天的平均创面愈合率分别为正常组 91.7%、M.E.组 79.6%、rhEGF 组 71%、凡士林组 55.1%和糖尿病空白组 43.3%。形态学染色显示,M.E.组较空白组和凡士林组有更活跃的肉芽组织形成、再上皮化和新生血管形成。糖尿病大鼠中 p-STAT3 核易位减少,Bcl-2、CyclinD1 和血管内皮生长因子(VEGF)表达下调,rhEGF 和 M.E.可改善这一现象。
蛆提取物可能是治疗糖尿病皮肤创面的一种替代和/或辅助候选药物,因为它能增强 STAT3 的激活。