Diosmetin Substituent Impact On Venous Thrombosis Proteome And Research Of Target Protein

Galium verum L. is a wild plant from Northeast China, this grass is well known as an excellent food source of vitamin, and it also contains phytochemicals such as Chlorogenic Acid, Salicylic Acid and flavone. Anti-venous thrombosis activity values obtained from Galium were higher than other plant particuLarly rich in flavones. Diosmetin substituent compounds by high performance liquid chromatography (RPLC), as main factor responsible for anti-venous thrombosis activity,were determined. This study was to analyze the proteome changes of diosmetin substituent in venous thrombosis of plasma ,in order to search for biomarkers and target proteins in venous thrombosis, as will as a lead compound and provide a theoretical basis.To investigation the proteome changes of the plasma, Wistar rats were selected to establish the model and verificated by pathologic diagnosis and protein C according to it's key role in thrombosis. The control group did not undergo any processing, the model group and the prevention group were established thrombosis model in vitro and the difference between them was the prevention group had been given diosmetin substituent before model was established. Dimensional gel electrophoresis and high performance liquid chromatography method were combined to analysis proteome. Plasma proteins were extracted by high velocity centrifugation and separated by Sephadex column to remove high-abundance proteins such as immunoglobuLin. Low-abundance proteins were separated by dimensional gel electrophoresis and the images were analyzed by PDquest software.Comparative and analysis dimensional gel electrophoresis gel images of the control group, the model group and the prevention group, there are 147 protein spots in model group,while 99 protein spots in prevention group. Comparing with the model group there are 49 expressed differential protein spots in the prevention group ,among them there 28 protein spots expressing increased while 21 decreased ; 56 protein spots were identified in model group only, 8 differently proteins spots were in prevention group only.By Matrix-Assisted Laser Desorption/ Ionization Time of Flight Mass Spectrometry, 13 kinds (10named , 3unnamed )proteins were identified successfuLly , the higher rate matched target protein are Full = Serum albumin; Flags: Precursor, serum albumin precursor, glutathione S-transferase pi, albumin isoform CRA_a. The retrieval scores are 287,278,109,104, and the matching rate are 25%, 23%, 40%, 37% respectly.