Clinopodium tomentosum (Kunth) Govaerts is an endemic species in Ecuador, where it is used as an anti-inflammatory plant to treat respiratory and digestive affections. In this work, effects of a Clinopodium tomentosum ethanolic extract (CTEE), prepared from aerial parts of the plant, were investigated on vascular endothelium functions. In particularly, angiogenesis activity was evaluated, using primary cultures of porcine aortic endothelial cells (pAECs). Cells were cultured for 24 h in the presence of CTEE different concentrations (10, 25, 50, and 100 μg/ml); no viability alterations were found in the 10-50 μg/ml range, while a slight, but significant, proliferative effect was observed at the highest dose. In addition, treatment with CTEE was able to rescue LPS-induced injury in terms of cell viability. The CTEE ability to affect angiogenesis was evaluated by scratch test analysis and by an in vitro capillary-like network assay. Treatment with 25-50 μg/ml of extract caused a significant increase in pAEC’s migration and tube formation capabilities compared to untreated cells, as results from the increased master junctions’ number. On the other hand, CTEE at 100 μg/ml did not induce the same effects. Quantitative PCR data demonstrated that FLK-1 mRNA expression significantly increased at a CTEE dose of 25 μg/ml. The CTEE phytochemical composition was assessed through HPLC-DAD; rosmarinic acid among phenolic acids and hesperidin among flavonoids were found as major phenolic components. Total phenolic content and total flavonoid content assays showed that flavonoids are the most abundant class of polyphenols. The CTEE antioxidant activity was also showed by means of the DPPH and ORAC assays. Results indicate that CTEE possesses an angiogenic capacity in a dose-dependent manner; this represents an initial step in elucidating the mechanism of the therapeutic use of the plant.

中文翻译：

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绒毛次要叶（Kunth）Govaerts叶提取物对猪主动脉内皮细胞原代培养的体外细胞增殖和血管生成的影响。

绒毛毡头草（Kinoth）Govaerts是厄瓜多尔的特有种，被用作治疗呼吸道和消化道疾病的抗炎植物。在这项工作中，研究了从植物的空中部分制备的金毛茶（Clinopodium tomentosum）乙醇提取物（CTEE）对血管内皮功能的影响。特别是，使用猪主动脉内皮细胞（pAEC）的原代培养评估了血管生成活性。细胞在CTEE不同浓度的存在下培养24小时（10，25，50，和100  μ微克/毫升）; 无活力的改变在10-50发现 μ在最大剂量范围内，虽然有微弱但显着的增生作用。此外，就细胞活力而言，CTEE治疗能够挽救LPS诱导的损伤。通过划痕试验分析和体外毛细管样网络测定法评估CTEE影响血管生成的能力。用25-50治疗 μ微克/毫升提取物引起的与未处理细胞相比在PAEC的迁移和管形成能力的显著增加，如由增加的主结号码的结果。在另一方面，在CTEE 100  μ微克/毫升未诱导相同的效果。定量PCR数据证实，FLK-1 mRNA的表达在25的CTEE剂量显著增加 μ克/毫升 通过HPLC-DAD评估CTEE的植物化学组成；酚酸中的迷迭香酸和类黄酮中的橙皮苷是主要的酚类成分。总酚含量和总黄酮含量测定表明，黄酮是多酚类中含量最高的一类。还通过DPPH和ORAC分析显示了CTEE的抗氧化活性。结果表明，CTEE具有剂量依赖性的血管生成能力。这是阐明植物治疗用途机理的第一步。