Diosmin is the aglycone moiety of diosmetin (3',5,7 trihydroxy-4'methoxy radicals flavonoids), a naturally occurring flavone glycoside, whose antithrombotic effect was studied in rats. This study was designed to find the protein changes of venous thrombosis in Wistar rats comparing conditions with and without Diosmin treatment by two-dimensional gel electrophoresis (2-DE), and investigate the effect of a crucial protein known as CEP350 on human vascular endothelial cell growth. Through prior chromatographic purification with macroporous absorption resin (AB-8) and polyamide, Diosmin was isolated from Galium verum L. by solvent extraction, then purified to 98% purity using HPLC. Wistar rats were divided into control group, model group, and prevention group. And their venous thrombosis tissue segments were dissected and prepared for histopathological examination and detection of plasma protein C (PC). Next, proteomic analysis was performed with the samples. Low-abundance proteins of the three groups were separated by two-dimensional gel electrophoresis (2-DE). 2-DE analysis revealed that 191 protein spots were differentially expressed among those three groups. For protein identification, we selected six spots to use matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) detection, and then do the homology search in NCBI database. Considering characteristics of these proteins, we proposed CEP350 is related to spindle assembly. Furthermore, we used Lipofectamine 2000 to transfect HUVECs with CEP350 siRNA and evaluated the extent of silencing using real time-polymerase chain reaction (RT-PCR). Cells were stained for immunofluorescence with tubulin-tracker red, and structural changes were analyzed using laser scanning confocal microscope. We concluded that CEP350 depletion decreased microtubule stability. Dosmin could modulate the assemble of spindle from unevenly distributing and protect body from varicose veins by regulating CEP350.
Keywords: CEP350; Centromere; Diosmin; Proteomics; Spindle; Two-dimensional gel electrophoresis.
Copyright © 2017. Published by Elsevier B.V.