Effect Of 10-hydroxy-2-decenoic Acid On Vascular Smooth Muscle Cells Injury And Erythrocyte Membrane Fluidity

The objective of this study was to explore the effects of 10-hydroxydecin-2-enoic acid（10-HDA）on mouse vascular smooth muscle cells.The molecular mechanism of the anti-damage effect of VSMCs on the fluidity of erythrocyte membrane and its effect on the fluidity of erythrocyte membrane were clarified.This study is of great significance to reveal more biological functions of this drug and to treat diseases such as hypertension and atherosclerosis.At the same time,the effect of 10-HDA on the fluidity of erythrocyte membrane was further explored to provide theoretical basis for the treatment of diseases related to the fluidity of erythrocyte membrane.In this study,a mouse VSMCs damage model was established,and the control group,·OH damage group and·OH+10-HDA group were set up.The molecular mechanism of 10-HDA on·OH damage was analyzed by cell morphological observation,bioinformatics analysis,MTT,Western blot and other experimental techniques.Using mouse red blood cells as experimental materials,Using high performance liquid chromatography（HPLC）method,flow cytometry,cell membrane far infrared fluorescent probe(1,1’-Dioctadecyl-3 filling’,3’-Tetramethylindodicarbocyanine,4-Chlorobenzenesulfonate Salt,The effect of 10-HDA on the fluidity of erythrocyte membrane in mice was investigated by Di D staining.In this study,it was found that in injured VSMCs,the scavenging rate of 10-HDA for·OH was greater than 20%,and 10-HDA significantly enhanced the activity of VSMCs injured by·OH,and significantly inhibited the production of malondialdehyde（MDA）,a marker of cell lipid peroxidation.Glutathione（GSH）level in·OH treated VSMCs was increased.By proteomic analysis,195 proteins co-regulated by·OH and 10-HDA were down-regulated and then up-regulated.These proteins are mainly involved in important life activities such as protein synthesis（such as translation,protein transport,ribosome and RNA binding）and energy metabolism（such as fatty acid degradation,glycolysis and gluconogenesis）.Through dyeing found 10-Di D HDA can enter the red cell membrane,by density functional theory suggests that 10-HDA may with the main composition of membrane lipid bilayer lecithin（phosphatidylcholine,PC）and cephalin（phosphatidylethanolamine,PE）interaction,This may be how the 10-HDA enters the membrane.According to cell morphological observation,MDA and erythrocyte membrane fluidity experiment,10-HDA can increase the fluidity of erythrocyte membrane and improve the membrane fluidity decreased by·OH,which may be related to the decrease of lipid peroxidation of erythrocyte membrane caused by·OH by 10-HDA.Dissolved oxygen experiment showed that 10-HDA can increase the release of O₂in red blood cells,which may be related to the improvement of erythrocyte membrane fluidity.At the same time,Western blot showed that some related proteins in erythrocyte membrane,such asβ-actin and band 4.1 protein（EPB41）,were not affected by 10-HDA treatment,indicating that 10-HDA mainly acted on the lipid part of the cell membrane.