Studies On The Effect Of Sodium Arsenite On The Migration And Proliferation Of HUV-ECs Its Mechanism

Many studies have shown that the surrounding blood vessels is a major target organ toxicity for arsenic. The normal function of peripheral vascular exercise depends on the maintenance of endothelial integrity and damage repaired in time. Vascular endothelial cells play a decisive role in this process. In order to study Arsenic compounds for vascular endothelial cell damage repair function of its molecular mechanism. Experiment by cell damage repair and cell proliferation experiments were studied sodium arsenite on cell migration and cell proliferation capacity, and to explore arsenicals on HUVECs migration and proliferation and its molecular mechanism in order to look forward to arsenic disease molecular mechanism of vascular injury provides a new explanation for disease prevention and treatment of arsenic to provide a new basis.Methods:1.Culture Vascular endothelial cells:We recovery HUVECs cells which stored in liquid nitrogen.Containing 10% serum IMDM medium placed in 37℃, 5% CO2 incubator temperature.2.Detection of cell migration:(1).Cell damage (scratch) test for the role of cell sodium arsenite cell migration after 24h.(2).Inverted phase contrast microscope HUVECs morphology.The formula for cell migration inhibition: Scratch width of the cells after incubation/ Pre-incubation of cells×100% scratch width3.Detection of cell proliferation: (1) With CCK-8 kit detection different concentration of cells HUVECs cells by a series of concentrations of sodium arsenite solution .continuous observation of 4d.(2) calculated according to the formula of sodium arsenite inhibit the rate of cell proliferation.4. With real-time PCR (RT-PCR) detection of migration-related and proliferation-related genes expression.Results:Inverted phase contrast microscope observation of morphological changes in vascular endothelial cells.The results showed that :Vascular endothelial cells do not differ from normal endothelial when after low concentrations of sodium arsenite. However,with the increasing concentration of sodium arsenite. Cells from flat polygonal cell become irregular type or circ ular. And can be seen a large dead cells.Sodium arsenite in the context of the study can cause changes in cell migration. As the concentration of sodium arsenite solution increased, decreased cell migrationSodium arsenite can induce changes in cell proliferation. The measurement can be obtained: Sodium arsenite can inhibit human umbilical vein endothelial cell proliferation, and with the increase of arsenic concentration more significantly inhibited cell proliferation.Real-time quantitative PCR results showed that:(1) migration-related genes: Sodium arsenite can inhibit FAK expression and increases MMP-9 expression.(2) proliferation-related genes: Sodium arsenite can inhibit the expression of C-MYC.Experimental results:Sodium arsenite can be inhibited HUVECs cell proliferation and migration capabilities.It was first discussed in arsenic disease caused by vascular injury and cell proliferation, migration relationship. Experimental results show that arsenic on vascular endothelial cell injury and repair function through decreased proliferation and migration of endothelial cells to achieve.In the experiment, we found that sodium arsenite can be reduced FAK expression and increased expression of MMP-9 to inhibit vascular endothelial cell migration; And by reducing the expression of C-MYC inhibits endothelial cell proliferation. In view of these gene products were in cell migration and proliferation play an important role, suggesting that inhibiting the expression of these genes were sodium arsenite inhibits cell migration and proliferation of important molecular mechanism.Sodium arsenite inhibits cell migration and proliferation of HUVECs. They are showed as a typical dose-response relationship. Prompted by sodium arsenite to inhibit vascular endothelial cell migration and proliferation of vascular injury site, and then damage its vascular repair. Further studies showed that sodium arsenite could inhibit FAK and C-MYC gene expression and increased expression of MMP-9 gene. In view of these gene products in cell migration and proliferation are important role, suggesting that inhibiting the expression of these genes were sodium arsenite injury cell migration and proliferation of important molecular mechanism.