Pro-angiogenic actions of Salvianolic acids on in vitro cultured endothelial progenitor cells and chick embryo chorioallantoic membrane model.
OBJECTIVE
Salvia miltiorrhiza (SM, also known as DanShen) is one of the well-known widely-used Chinese herbal medicines in clinical practice. In this study we aimed to demonstrate the pro-angiogenic effects of Salvianolic acids (SAs) to treat illnesses such as ischemic cardiovascular diseases, the main active components of aqueous extract of SM.


METHODS
To do this, new-born rat spleen mononuclear cells were isolated and endothelial progenitor cells (EPCs) were expanded (not more than 24h) SD. Then the pro-angiogenic activities of SAs were evaluated on in vitro cultured EPCs and chick embryo chorioallantoic membrane (CAM) model. And the adherent cells were stained with DiI complexed acetylated low-density lipoprotein (DiI-acLDL) and fluorescein Ulex Europaeus agglutinin-1 (FITC-UEA-1), and then viewed by laser scanning confocal microscope (LSCM) to confirm EPCs lineage. EPCs identification was also tested by ultrastructural analyses. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, transwell chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating those on fibronectin-coated dishes, and then counting adherent cells.


RESULTS
EPCs phenotype was confirmed by the presence of double positive cells for DiI-acLDL uptake and lectin binding and identification of Weibel-Palade body in cytoplasm by ultrastructural analyses. Incubation of EPCs with SAs increased the number of EPCs and promoted EPCs migratory, adhesive and in vitro vasculogenesis capacity. SAs also promoted angiogenesis as evidenced by CAM model.


CONCLUSIONS
The results of the present study suggest that SAs may have utility for therapeutic postnatal vasculogenesis of ischemic tissue, contributing to the clinical benefit of SM therapy in patients with coronary artery disease.
Li
Yu-Juan
YJ
China Academy of Chinese Medical Sciences, Xiyuan Hospital, Beijing, PR China.
Duan
Chang-Ling
CL
Liu
Jian-Xun
JX
Xu
Yong-Gang
YG
D000818:Animals; D000831:Animals, Newborn; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000187:drug effects / Q000378:metabolism; D004365:Drugs, Chinese Herbal / Q000494:pharmacology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D018613:Microscopy, Confocal; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010936:Plant Extracts / Q000494:pharmacology*; D051381:Rats; D027561:Salvia miltiorrhiza / Q000737:chemistry*; D013154:Spleen / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D013234:Stem Cells / Q000187:drug effects* / Q000378:metabolism
D004365:Drugs, Chinese Herbal; D010936:Plant Extracts