ABSTRACT
<p><b>OBJECTIVE</b>To investigate the function of VEGF pathway in promoting angiogenesis with Xuefu Zhuyu Tang (XFZYT).</p><p><b>METHOD</b>Serum pharmacology technique was adopted to treat endothelial cells ECV304 with XFZYD containing serum and blank serum with concentrations of 1.25%, 2.5% and 5%. Methyl thiazolyl tetrazolium (MTT) assay, boyden chamber migration assay and in vitro vessel formation assay were used to test endothelial cell proliferation, migration and angiogenesis capacities. ELISA, immunohistochemistry and RT-PCR were used to detect secretion and expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR2).</p><p><b>RESULT</b>XFZYT-CS with a concentration of 1.25% only affected angiogenesis capacity in vitro. XFZYT-CS with a concentration of 2. 5% promoted cell proliferation, migration and angiogenesis capacities and significantly increased VEGF level and VEGF/VEGFR2 expressions. XFZYT-CS with a concentration of 5% only up-regulated intracellular VEGF expression and thereby improving endothelial cell proliferation, migration and angiogenesis.</p><p><b>CONCLUSION</b>XFZYT can induce endothelial cell proliferation, migration and angiogenesis by up-regulating VEGF-VEGFR2 pathway, which partially proves its angiogenesis effects.</p>
Subject(s)
Animals , Female , Humans , Male , Rats , Cell Movement , Cell Proliferation , Drugs, Chinese Herbal , Pharmacology , Endothelial Cells , Cell Biology , Metabolism , Neovascularization, Physiologic , Rats, Sprague-Dawley , Receptors, Vascular Endothelial Growth Factor , Genetics , Metabolism , Vascular Endothelial Growth Factor A , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the angiogenesis modulation mechanism of Xuefu Zhuyu Decoction () on the endothelial cell line ECV304.</p><p><b>METHODS</b>ECV304 cells were treated with 2.5% Xuefu Zhuyu Decoction-containing serum (XFZYD-CS) for 24 h, 48 h or 72 h. Thiazolyl blue tetrazolium bromide (MTT), fluorescence activating cell sorter (FACS), migration, adhesion and in vitro tube formation assays were conducted to confirm an angiogenesis effect of XFZYD at 3 time points. An analysis of angiogenesis regulator profiles was performed at 3 times with real-time polymerase chain reaction (RT-PCR) superarray.</p><p><b>RESULTS</b>At 48 h, XFZYD-CS induced ECV304 significantly improved cell viability, number in S phase, migration, adhesion and tube formation. At 24 h and 72 h, only cell migration was elevated. Microarray results showed that the expression of 27 angiogenesis-related genes was changed.</p><p><b>CONCLUSION</b>XFZYD-CS treatment induced angiogenesis on ECV304 cells with significant cellcular changes occurring at 48 h and genetic changes as early as 24 h.</p>
Subject(s)
Humans , Angiogenesis Inducing Agents , Pharmacology , Cell Adhesion , Genetics , Cell Line , Cell Movement , Genetics , Cell Proliferation , Cell Survival , Genetics , Drugs, Chinese Herbal , Pharmacology , Endothelial Cells , Metabolism , Gene Expression Regulation , Neovascularization, Physiologic , Genetics , Oligonucleotide Array Sequence Analysis , MethodsABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of Xuefu Zhuyu Decoction ()-containing serum (XFZYD-CS) on endothelial progenitor cell (EPC) tube formation in vitro.</p><p><b>METHODS</b>Mononuclear cells from rat bone marrow were prepared in a Ficoll density gradient centrifuge. EPCs were separated by the differential attachment method, and observed with inverted microscope for the effect of XFZYD-CS on EPC tube formation.</p><p><b>RESULTS</b>After one day, EPCs exposed to the serum containing 5%, 10% and 15% XFZYD-CS formed typical tubes or vessel networks. The tube formation time was two days ahead of the control group and the size of most tubes in the serum groups was smaller than in the control group.</p><p><b>CONCLUSION</b>XFZYD-CS could induce EPC angiogenesis and hasten tube formation, especially in capillary vessels. The study provides experimental evidence for the plausibility of Xuefu Zhuyu Decoction in the treatment of ischemic diseases.</p>
Subject(s)
Animals , Rats , Angiogenesis Inducing Agents , Pharmacology , Cell Differentiation , Cell Shape , Physiology , Cells, Cultured , Drug Evaluation, Preclinical , Drugs, Chinese Herbal , Pharmacology , Endothelial Cells , Cell Biology , Physiology , Neovascularization, Physiologic , Rats, Sprague-Dawley , Stem Cells , Cell Biology , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To study the acting mechanism of endothelial cell line ECV304 in regulating angiogenesis induced by Xuefu Zhuyu Decoction (XFZY).</p><p><b>METHODS</b>The angiogenesis effect of XFZY-contained serum (XFZY-CS) was confirmed by observing its impact on proliferation, cell cycle, migration of ECV304 and on vascular neogenesis in vitro. Then the effect of XFZY on various angiogenesis controlling factors was analyzed with gene chip microarray technique.</p><p><b>RESULTS</b>Treatment of XFZY-CS in 2.5% concentration for 48 h showed evident actions of enhancing ECV304 activity, increasing cell numbers of S phase, inducing cell migration and promoting the in vitro angiogenesis. Meanwhile, expressions of four angiogenesis controlling genes were up-regulated and 10 were down-regulated.</p><p><b>CONCLUSION</b>The angiogenesis mechanism of ECV304 induced by XFZY is complex, it shows a multi-pathway and multi-target feature.</p>