Zeng Zhi, Zhu Bang-Hao
Department of Pharmacology, Sun Yat-sen University, Guangzhou, 510080, China.
Department of Pharmacology, Sun Yat-sen University, Guangzhou, 510080, China.
J Ethnopharmacol. 2014 Jul 3;154(3):653-62. doi: 10.1016/j.jep.2014.04.038. Epub 2014 May 1.
Zicao is a traditional wound healing herbal medicine that has been used for several hundred years in China. A survey of the published literatures revealed that arnebin-1, one of the naphthoquinone derivatives, played the most important role in wound healing property of this plant. However, whether arnebin-1 affects angiogenesis in vitro and has an effect on wound healing process in diabetic rats remains enigmatic. To investigate the effect of arnebin-1 with or without VEGF on proliferation, migration and tube formation of HUVECs in vitro and the effect of its topical application in the form of ointment on wound healing in a cutaneous punch wound model of alloxan-induced diabetic rats in vivo.
The pro-angiogenic functions of arnebin-1 on HUVECs including proliferation, migration and angiogenesis were evaluated through MTT assay, wound healing assay, transwell assay and tube formation assay in vitro. Male Sprague-Dawley rats were injected intraperitoneally with alloxan to induce type І diabetic rats. Three wounds were created in each rat on the dorsal surface, and then divided to be basement treated, arnebin-1 ointment treated and untreated group correspondingly. The indicators including wound closure rate and histological evaluation were investigated on day 4 and 7 post-wounding.
Without VEGF, arnebin-1 did not affect the proliferation of HUVECs significantly, but had a positive effect on cell migration and tube formation. However, in the presence of minimal VEGF, Arnebin-1 could increase the proliferation, enhance the migration and promote the tube formation of HUVECs significantly. The wound closure rate was increased significantly in arnebin-1 treated group compared to that of untreated and basement treated groups in diabetic rats, and the histological evaluation also showed well organized dermal layer, reduced number of macrophages, increased number of fibroblasts, remarkable degree of neovascularization and epithelization in arnebin-1 treated group.
These findings suggest that arnebin-1 has a pro-angiogenic effect, and a synergetic effect with VEGF promotes the wound healing process in diabetic rats.
紫草是一种传统的伤口愈合草药，在中国已使用了数百年。对已发表文献的调查显示，萘醌衍生物之一的arnebin-1在该植物的伤口愈合特性中起最重要作用。然而，arnebin-1是否在体外影响血管生成以及对糖尿病大鼠的伤口愈合过程是否有影响仍然未知。为了研究arnebin-1单独或与血管内皮生长因子（VEGF）联合应用对体外人脐静脉内皮细胞（HUVECs）增殖、迁移和管腔形成的影响，以及其软膏剂型局部应用对四氧嘧啶诱导的糖尿病大鼠皮肤打孔伤口模型体内伤口愈合的影响。
通过MTT法、伤口愈合试验、Transwell试验和体外管腔形成试验评估arnebin-1对HUVECs的促血管生成功能，包括增殖、迁移和血管生成。雄性Sprague-Dawley大鼠腹腔注射四氧嘧啶诱导Ⅰ型糖尿病大鼠。在每只大鼠的背部制造三个伤口，然后相应地分为空白对照组、arnebin-1软膏治疗组和未治疗组。在受伤后第4天和第7天观察伤口闭合率和组织学评估等指标。
在没有VEGF的情况下，arnebin-1对HUVECs的增殖没有显著影响，但对细胞迁移和管腔形成有积极作用。然而，在存在微量VEGF的情况下，arnebin-1可显著增加HUVECs的增殖、增强迁移并促进管腔形成。与糖尿病大鼠的未治疗组和空白对照组相比，arnebin-1治疗组的伤口闭合率显著提高，组织学评估还显示arnebin-1治疗组的真皮层组织良好、巨噬细胞数量减少、成纤维细胞数量增加、新生血管形成和上皮形成程度显著。
这些发现表明，arnebin-1具有促血管生成作用，并且与VEGF协同作用可促进糖尿病大鼠的伤口愈合过程。
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