| Background:Most of the atherosclerotic plaques are located in places where blood flow is slightly slower,such as bifurcation,bending,and stenosis.Research shows that,due to the different shear force and the result is different also,and in most of the blood vessels,buoyed by the high shear force,also is our common high shear force,recent research conclusion is relatively consistent,it is generally believed high shear force-with the role of atherosclerosis formation.In the case of vascular bifurcation,bending and so on,it is easy to produce vortexes,so that the flow direction is uneven,and the velocity is affected,which is significantly lower than the previous position.(0± 4 dyne/cm2).Exosomes is a kind of size between 30-100nm vesicles with the active cell secreted release,which specific loaded nucleic acid sequences,protein and other ingredients derived cells,and selectively transfer to target cells and produce corresponding biological functions in target cells.The largest characteristic of Exosomes,which is different from other signaling pathways,is the ability to carry nucleic acid sequences,including miRNA.Objective:In order to explore whether Exosomes participates in the mechanism of causing AS,the effect of blood flow shear force on exocrine secretion is explored.Methods:Through the neonatal umbilical cord,HUVECs was extracted and cultured,and the above cells were treated with different time shear forces and separated by overspeed centrifugation.The total protein and exocrine protein were measured by BCA.The expression level of the endogenous protein in the exudate was detected by Western blot,and whether the secretion of exocrine was different under different shear force was measured.Result:1)Western blot showed the particle has TSG101,CD63 protein.Confirm that the vesicles were exosomes.2)After low shear stress,the number of exocrine increased significantly with time.The peak concentration at 24 hours reached a peak,which was significantly different from that in the control group.3)The longer the stimulation time was,the more the exocrine secretion was.The secretion of exocrine was positively correlated with the stimulation time at low shear stress.Conclusion:1)Ultracentrifoge md sucrose gradient centrifiigatiori can effectively extract exosomes from HCMV particles.2)oscillatory shear stress increased the expression of exosomes in endothelial cells.Background:In recent years,with the development of its research,the characteristics and functions of exocrine are gradually recognized.Exocrine is a substance carrying many kinds of signal transduction functions,including protein,Messenger RNA(mRNA),tiny RNA(microRNA)and so on,which mediate cell to cell communication.Exocrine is mainly formed,secreted and released in the endocytic endocytosis system.Objective:The exocrine mainly plays a role in transmitting signals,and the specific effect depends on the role of different miRNA.Whether the blood flow shear force can enhance the role of miRNA and increase the role of the secretion of the above factors.At the same time,which kind of miRNA can be changed by the stimulation of low shear force.In this study,the cultured endothelial cells were stimulated by low shear force and could be observed by immunofluorescence to induce the changes of genes in the exocrine body.Methods:HUVECs was extracted from the umbilical cord of neonates and cultured.,the cells were exposed to oscillatory shear stress(0±4 dyne/cm2).The cells of the control group were maintained in a static state.he total RNA was extracted from the exocrine cells of different groups,and the difference in the expression of miRNA was revealed by the fluorescence quantitative method.Result:Compared with the static control group,the low shear force stimulation group significantly increased the expression of miRNA-145,miRNA-143 and other genes.Conclusion:1)The exocrine carries out signal transduction through its miRNA,and then regulates cell migration and proliferation.2)The difference of miRNAs expression in the exocrine after shear force is greatly different from the shear force.Background:Most vulnerable plaques occur in the vascular segments that continue to be low in ESS.Moreover,the degree of low ESS on the baseline is closely related to the severity of the high risk plaque.Previous studies have observed that low shear stress induced increased secretion of exocrine in endothelial cells and altered the signal transduction mechanism in the exocrine body.Objective:In this part it will extract the low shear exosome exosome gels after stimulation with the stationary state of the coronary artery smooth muscle cells were transfected in vitro to observe their study,coronary artery smooth muscle cells by immunofluorescence and Western blot method proved to be not caused by the increment and migration of coronary artery smooth muscle cells.Methods:In the fluorescence staining of exosomes with coronary artery smooth muscle endothelial cells after incubation by fluorescence confocal microscope observation of exosome uptake of hCASMCs effect,and through the MTT method to detect the hCASMCs value of the level of Transwell and Western migration assay of hCASMCs level detection of hCASMCs related protein blot phenotypeResult:1)Fluorescent confocal microscopy can be used to observe the effective uptake of exocrine by coronary artery smooth muscle cells.2)Compared with both control groups and blank group,the experimental group could significantly increase HCASMC migration function(p<0.05).3)Compared with both control groups and blank group,After the intake of the exocrine in the low shear stress treatment group,the value added of the coronary artery smooth muscle cells increased significantly.4)Compared with both control groups and blank group.There were differences in Osteopontin,Epiregulin and Elasti protein between coronary artery smooth muscle endothelial cells.But A-SMA,Caldesmon and Calponin protein had little difference.Conclusion:1)Fluorescence confocal microscopy confirmed that hCASMCs really can absorb the exocrine..2)Exosomes treated with low shear stresscould significantly increase HCASMC migration function.3)Exocrine in the low shear stress added of the coronary artery smooth muscle cells increased significantly.4)Exosomes treated with low shear stress had significant difference in the changes of hCASMCs proliferation and protein phenotype.Background:Through previous studies,we found that low shear force can stimulate the production of more exocrine in the primitive endothelial cells.Through the incubation and culture of secretory exocrine and smooth muscle cells,it is observed that smooth muscle cells can fully absorb exocrine bodies,and then cause the next step of signal transduction.Different RNA plays different roles in signal transduction.What gene plays a major role in signal transduction?We must enhance it alone to confirm this.Objective:Separate miRNA-145 as a single research object,and through its inhibitor and negative control group were compared,observe whether the gene can enhance the value-added and migration of cells,and then can determine whether to promote the migration and proliferation of smooth muscle cells through the regulation of the gene.Methods:By buying the synthetic miR-145 mimic,inhibitor,and negative.Transfection of the external secretory cells and the efficiency of transfection according to the enzyme scale.Through the MTT method to detect the hCASMCs value of the level of Transwell and Western migration assay of hCASMCs level detection of hCASMCs related protein blot phenotype.Result:l)Three groups of miRNA-145,miRNA-145 inhibitors and negative control agents were cultured after multiple generation of cells.The transfection effect of miRNA-145 was significantly higher than that of the other 2 groups,and there was a statistically significant difference compared with the other two groups..2)Compared with inhibitor and negative,the experimental group could significantly increase HCASMC migration function(p<0.05).3)Compared with inhibitor and negative,After the intake of the exocrine in the low shear stress treatment group,the value added of the coronary artery smooth muscle cells increased significantly.4)Compared with inhibitor and negative.There were differences in Osteopontin,Epiregulin and Elasti,A-SMA,Caldesmon and Calponin protein between coronary artery smooth muscle endothelial cells.Conclusion:1)In the genes contained in the exocrine,the proliferation and migration of miRNA-145 were significantly improved..2)After inhibition of miRNA-145,other genes had no significant effect on the proliferation and migration of smooth muscle endothelial cells.3).the low shear force is mediated by regulating the proliferation,migration and phenotypic transformation of coronary artery smooth muscle cells by regulating the specific miRNA in the exocrine secreted by HUVECs. |
PDF Full Text Request