Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Banihashem Sq., Banihashem St., Resalat Highway, P.O. Box: 16635-148, Tehran, Iran.
Department of Horticultural Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran.
Daru. 2020 Dec;28(2):533-543. doi: 10.1007/s40199-020-00356-7. Epub 2020 Jul 1.
Angiogenesis related abnormalities underlie several life-threatening disorders. Despite approved therapies, scientists have yet to develop highly efficient, low cost approaches with minimal side effects.
We evaluated the antiangiogenic activity of 50% hydroalcoholic extracts of Pergularia tomentosa L. root and aerial parts along with their EtOAc and water fractions, in vivo and in vitro. Transgenic zebrafish line Tg(fli1:EGFP) was used for in vivo assay and human umbilical vein endothelial cell (HUVEC) migration test along with possibility of tube formation were performed as in vitro tests. Furthermore, microvasculature in chicken chorioallantoic membrane (CAM) was assessed under P. tomentosa treatment. The fractionation of the 50% hydroalcoholic extracts was led to the identification of the best active fraction in this study. The metabolite profiling of the active fraction was also carried out using LC-HRESIMS analysis.
Pergularia tomentosa markedly inhibited intersegmental vessel (ISV) formation at 48 h post-fertilization (hpf) embryos in zebrafish. The water fraction of root hydroalcoholic extract (PtR2), showed strong antiangiogenic effect with minimal adverse viability impacts. Over 80% of embryos showed more than 50% inhibition in their ISV development at 20 and 40 μg/mL. PtR2 at 20 μg/mL substantially reduced human umbilical vein endothelial cell (HUVEC) migration up to 40%, considerable destruction of the formed tubes in the tube formation and microvasculature in CAM assays. Immunocytochemistry showed a marked reduction in vascular endothelial cadherin (VE-cadherin) abundance at cell junctions concurrent with substantial reduction of phospho-Akt (p-Akt) and β-catenin protein expressions. Phytochemical profile of PtR2 showed a rich source of cardenolide structures, including ghalakinoside, calactin and calotropin derivatives.
Thus, the P. tomentosa cardenolide-rich fraction (PtR2) may hold a considerable promise for an antiangiogenic impact by impairment of endothelial cell (EC) migration and viability. Graphical abstract.
血管生成相关异常是几种危及生命的疾病的基础。尽管有批准的治疗方法，但科学家们尚未开发出高效、低成本且副作用最小的方法。
我们评估了 Pergularia tomentosa L. 根和地上部分的 50%水醇提取物及其 EtOAc 和水馏分的体内和体外抗血管生成活性。转基因斑马鱼 Tg(fli1:EGFP) 用于体内测定，人脐静脉内皮细胞 (HUVEC) 迁移试验和管形成可能性作为体外试验进行。此外，还评估了 Pergularia tomentosa 处理下鸡绒毛尿囊膜 (CAM) 的微血管。50%水醇提取物的分级导致确定了本研究中最佳活性部分。还使用 LC-HRESIMS 分析对活性部分进行了代谢物分析。
Pergularia tomentosa 显著抑制了斑马鱼受精后 48 小时胚胎的节间血管 (ISV) 形成。根水醇提取物的水馏分 (PtR2) 表现出强烈的抗血管生成作用，对生存能力的不良影响最小。在 20 和 40 μg/mL 时，超过 80%的胚胎在其 ISV 发育中表现出超过 50%的抑制。20 μg/mL 的 PtR2 可将人脐静脉内皮细胞 (HUVEC) 迁移减少 40%，在管形成和 CAM 试验中的微脉管系统中形成的管的破坏相当大。免疫细胞化学显示血管内皮钙粘蛋白 (VE-cadherin) 在细胞连接处的丰度明显减少，同时磷酸化 Akt (p-Akt) 和 β-连环蛋白蛋白表达也明显减少。PtR2 的植物化学图谱显示出丰富的卡烯内酯结构来源，包括 ghalakinoside、calactin 和 calotropin 衍生物。
因此，P. tomentosa 富含卡烯内酯的部分 (PtR2) 可能通过损害内皮细胞 (EC) 的迁移和活力，具有相当大的抗血管生成潜力。
