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Role Of Reactive Oxygen Species In The Gene Expression Of ECV304 Proliferation

Posted on:2005-03-24Degree:MasterType:Thesis
Country:ChinaCandidate:L X XieFull Text:PDF
GTID:2144360125455044Subject:Pathology and pathophysiology
Abstract/Summary:PDF Full Text Request
[Objective] To explore role of reactive oxygen species(ROS) in the gene expression of ECV304 proliferation [Methods] The lines of human umbilical vein endothelial cell(ECV304) cultured in vivo were divided into three groups which were treated by Ang II .Ang II +N-acetyl-L-cysteine(NAC), and normal culture medium. First we observed the proliferous effect of ECV304 induced by Ang II at different concentration at 4th ,12th and 241 hour respectively with MTT and microscope. Then the contents of ROS('OH and NO) in three groups were detected by spectrophotometer. Finally we detected the related gene expression in the process of ECV304 proliferation mediated by ROS via the technique of genechips. [Results] (1)ECV304 incubated with Ang II (0.03125-1 umol/L ) for 12 hours increase the proliferation rate (PO.05 vs. NAC group and PO.05 vs. Control group). The effect of Ang II (0.0625μmol/L) on ECV304 reached maximum at 12th hour. NAC(10mmol/L) can counteract this effect. (2)It is significant that the negative correlation between proliferation rate and the contents of ROS(OH) (r - -0.8, PO.01).(3)The technique of genechips analysis suggests that the genes expression related to proliferation such as ERK, Akt, CCN and PCNA increase while the gene expression related to apoptosis such as DR6, Caspases6 BAK1 and PDCD8 decrease when ECV304 were induced by AngII(0.0625μmol/L) for 12 hours. In contrast, when ECV304 were induced by Ang 11(1 μmol/L) for 12 hours, we can obtain reverse results. [ Conlusions] ?1?ECV304 induced by Ang II can produce ROS(OH), and the contents of ROS('OH) increase with the prolongation of time and the enlargement of dose. ?2?It is significant that negative correlation between proliferation rate and the contents of ROS(OH). ?3?The target gene expression related to proliferation of ECV304 may be mediated by ROS(OH).?4?Antioxidant NAC can reduce the content of ROS(OH) and inhibit the proliferation of ECV304 induced by Ang II. (5) ROS(OH) may be the mocleculars which play an important role in the signal transduction and gene expression of proliferation.
Keywords/Search Tags:Ang II, ECV304, ROS, proliferation, gene expression
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