Effect Of Sargassum Fucoidan On Thrombus Include HUVEC And HMVEC

In this study,the crude polysaccharide was obtained by ultrasonic-assisted hot water extraction using Sargassum Henslowianum as raw material and named F.The components FD1,FD2 and FD3 were fractionally purified by DEAE C-52 column chromatography.The components FS1,FS2 and FS11 were purified by Sepharose CL-6B column chromatography on F and FD1.The chemical compositions of the above fucoidan components were determined.The results showed that the polysaccharide content,sulfate group content,fucose content and uronic acid content of each component were different.Infrared spectroscopy revealed that each component had a polysaccharide-characteristic functional group.The I-KI test and the Congo red test indicated that fucoidan has multiple complex branched structures,but only FS2 has a triple helix conformation.The monosaccharide composition and molecular weight of each fucoidan component were determined by HPLC.The results showed that each component contained almost no arabinose,FS2 was dextran,and the characteristic monosaccharides of other components were fucose.The molecular weight of each component ranges from 1.06×10⁵ to 3.14×10⁵ Da.The fucoidan with different purification degrees was prepared by the separation and purification steps,and the antithrombotic effect was studied by acting on two thrombus formation pathways.The model of mouse black tail thrombus was established to investigate the antithrombotic effect of each purified component in vivo.Human Umbilical Vein Endothelial Cells?HUVEC?and Human lung Micro-Vascular Endothelial Cells?HMVEC?were cultured to investigate the antithrombotic effects of each purified component on these two vascular endothelial cells in vitro.The anti-thrombosis effect of F,FD1 and FS1 was evaluated by establishing a model of Carrageenan induced mouse black tail thrombus,and the clotting time of each group of mice was determined by capillary method.The results showed that the black tail length of mice in each dose group of F,FD1 and FS1 was shorter than that of the model control group at the same time at 24 h,48 h and 72 h after model establishment,and FD1 and FS1 inhibited the black tail of mice.The effect was significantly higher than that of crude polysaccharide F?p<0.05,p<0.01?,and the 15mg/kg dose of FS1 had the best effect on inhibiting black tail?p<0.01?.The clotting time of the mice can prolong the clotting time of the mice,and the effect from high to low is FS1>FD1>F.To establish a Oxidized Low-density Lipoprotein induced cell injury model to investigate the protective effects of Sargassum fucoidan on Human Umbilical Vein Endothelial Cells?HUVEC?and Human lung Micro-Vascular Endothelial Cells?HMVEC?.Oxidized Low-density Lipoprotein can significantly?p<0.01?damage both vascular endothelial cells and cause a decrease in proliferation rate,and each fucoidan group can significantly protect cells from Oxidized Low-density Lipoprotein?p<0.01?.The effect of Oxidized Low-density Lipoproteins maintains the cell proliferation rate at normal cell levels.The secretion of ThromboxaneA₂?TXA₂?and Prostacyclin?PGl₂?in the two cells was determined by kit method.The results showed that the Sargassum fucoidan could not effectively regulate the secretion of TXA₂ and PGl₂ by HUVEC,But it can effectively promote the secretion of PGl₂ and inhibit the secretion of TXA₂ by HMVEC.