Diosmetin Impact On Venous Thrombosis And Research Of Target Protein

The diosetin extracted from Galium verum L. and the drug has an efficacy effect toinhibit cancer cell proliferation, reducing bleeding and pain. Besides, it has a good repairin radiation-induced oxidative damage; but research of diosetin mostly foucs on itssignificant role in venous thrombosis. Diosetin has been identified as an efficacy ofantithrombotic substances. This study analysis target proteins in venous thrombosis, andselect significantly changed proteins. Using umbilical vein endothelial cells to buildsensitive protein CEP350of siRNA reorganization, obtaining stable interfere proteinexpression in cell lines and detecting expression of target gene.Blood proteinsfrom pre-experiments in Wistar rats of control group, model groupand prevention group which were extracted of kit. Two-dimensional electrophoresis toseparation proteins and PDquest softwore was carried out to analysis images, and thenfinding differentially expressed proteins from the results. A total of307proteinsexpressed in the two-dimensional electrophoresis experiments. Prevention groupexpressed all the proteins and model group expressd91protein spots.137protein spotswere expressed just in the prevention group. Compared to the model group, protein2-fold increase points are32, and12protein2-fold down, selecting10differences inprotein matrix-assisted laser desorption time of flight mass spectrometry analysis andidentified50protein spots, including3unknown proteins.As CEP350which was identified in pre-research for target protein to silent thegenes by RNAi. Red microtubules fluorescent probe was used to stain both normal cellsand disrupt cells. The laser confocal microscope to observe changes of spindle andmicrotubules, at the same time RNA of disrupt cells was extracted to carry out reversetranscription and qPCR, in order to detect the expression of target genes to determine thesilence efficiency. Compare to normal cells, microtubles of interference group wasstretched and the number of spindle reduction in the distribution of cluster. RNAinterference indicates the primer was designed reasonable and the CEP350silent ratioachieved at68%. Combined these two conclutions, it is showed that CEP350play a rolein the process of spindle formation of cell division. And further description of thediosmetin affect the split of thrombosis endothelial cells through the regulation CEP350to influent the spindle assembly.Finally, take umbilical vein endothelial cells as object to optimize the dosingconcentration20μg/mL, the optimal dosing time of48h, and IC50is254.8μg/mL.