采用 3T3 成纤维细胞划痕实验测定金盏花提取物的创伤愈合效果。
Determination of the wound healing effect of Calendula extracts using the scratch assay with 3T3 fibroblasts.
机构信息
Department of Pharmaceutical Biology and Biotechnology, University of Freiburg, Freiburg, Germany.
出版信息
J Ethnopharmacol. 2009 Dec 10;126(3):463-7. doi: 10.1016/j.jep.2009.09.014. Epub 2009 Sep 23.
UNLABELLED
PHARMACOLOGICAL RELEVANCE: Presentation of the scratch assay as a convenient and inexpensive in vitro tool to gain first insights in the wound healing potential of plant extracts and natural compounds.
AIM OF THE STUDY
The present study deals with the optimization of the scratch assay which can be used as an in vitro model for quantification of fibroblast migration to and proliferation into the wounded area. It is suitable for the first evaluation of the wound re-epithelialization potential of crude herbal extracts, isolated compounds and pharmaceutical preparations. As a proof of concept three preparations from traditional medicinal plants were investigated.
MATERIALS AND METHODS
Swiss 3T3 albino mouse fibroblasts were used in monolayers and platelet derived growth factor as positive control. Hexane and ethanolic extracts from Calendula officinalis and Matricaria recutita, Hypericum oil as well as the triterpenoids faradiol myristate and palmitate were studied. To differentiate between proliferation and migration antimitotic mitomycin C was added.
RESULTS
Both extracts of Calendula officinalis stimulated proliferation and migration of fibroblasts at low concentrations, e.g. 10 microg/ml enhanced cell numbers by 64.35% and 70.53%, respectively. Inhibition of proliferation showed that this effect is mainly due to stimulation of migration. Faradiol myristate and palmitate gave comparable stimulation rates at an almost 50 microg/ml concentration, indicating that they contribute partially, but not most significantly to the wound healing effects of Calendula preparations. Extracts from Matricaria recutita were only moderately active. Hypericum oil was cytotoxic at concentrations higher than 0.5 microg/ml.
CONCLUSIONS
The scratch assay in the present form can be used as a promising scientific approach and platform to differentiate between plant extracts known for their wound healing and their anti-inflammatory properties.
目的
本研究旨在优化划痕试验，该试验可作为体外模型，用于定量评估成纤维细胞向创伤区域迁移和增殖的能力。它适用于初步评估粗草药提取物、分离化合物和药物制剂的伤口再上皮化潜力。作为概念验证，研究了三种来自传统药用植物的制剂。
材料和方法
使用瑞士 3T3 白化鼠成纤维细胞单层和血小板衍生生长因子作为阳性对照。研究了金盏花和母菊的正己烷和乙醇提取物、贯叶连翘油以及三萜脂肪酸酯法呢醇-myristate 和棕榈酸酯。为了区分增殖和迁移，添加了抗有丝分裂的丝裂霉素 C。
结果
金盏花的两种提取物在低浓度下刺激成纤维细胞的增殖和迁移，例如，浓度为 10μg/ml 时，细胞数量分别增加了 64.35%和 70.53%。增殖抑制表明，这种作用主要归因于迁移的刺激。法呢醇-myristate 和棕榈酸酯在几乎 50μg/ml 的浓度下给出了类似的刺激率，表明它们部分但不是最重要地促进了金盏花制剂的愈合效果。母菊的提取物仅具有中等活性。贯叶连翘油在浓度高于 0.5μg/ml 时具有细胞毒性。
结论
本研究形式的划痕试验可用作一种有前途的科学方法和平台，用于区分具有愈合和抗炎特性的植物提取物。
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