论文部分内容阅读
目的通过体外实验,研究H2O2对人脐静脉内皮细胞(ECV304)谷氧还蛋白(Grx)mRNA表达水平的影响。方法体外培养人脐静脉内皮细胞(ECV304);MTT实验观察H2O2对ECV304细胞增殖的影响;在时间相关性研究中,将细胞分为6组,第1组为正常对照组,第2、3、4、5、6组分别给予终浓度100μmol/L H2O2作用5min、10min、30min、60min、120min;在剂量相关性研究中,将细胞分为6组,第1组为正常对照组,第2、3、4、5、6组分别给予终浓度100μmol/L、200μmol/L、300μmol/L、400μmol/L、500μmol/L H2O2作用30min,采用Trizol一步法提取细胞总RNA,并用逆转录聚合酶链反应(RT-PCR)研究各组细胞Grx mRNA表达水平的差异。结果MTT结果表明,100μmol/L H2O2对细胞增殖无影响,200~500μmol/L H2O2均在不同程度上抑制细胞增殖(P<0.05)。RT-PCR结果显示,在时间相关性研究中,终浓度100μmol/LH2O2作用10min、30min、60min均能使ECV304细胞中GrxmRNA表达水平增高,30min达到最大值,差异显著(P<0.05);剂量相关性研究表明,终浓度100μmol/L H2O2~400μmol/L H2O2作用30min均能使Grx mRNA表达水平增高,差异显著(P<0.05),其中终浓度300μmol/L H2O2可使Grx mRNA的表达量最高(P<0.01)。结论H2O2影响人脐静脉内皮细胞Grx mRNA的表达水平。
Objective To investigate the effects of H2O2 on the expression of Grx mRNA in human umbilical vein endothelial cells (ECV304) in vitro. Methods Human umbilical vein endothelial cells (ECV304) were cultured in vitro. The effect of H2O2 on the proliferation of ECV304 cells was observed by MTT assay. In the time-dependent study, the cells were divided into 6 groups, the first group was normal control group, the second, 4, 5, 6 groups were given final concentration of 100μmol / L H2O2 for 5min, 10min, 30min, 60min, 120min; dose-related studies, the cells were divided into 6 groups, the first group of normal control group, The total RNA was extracted by Trizol one-step method in groups 3, 4, 5, and 6 for 30 min at final concentrations of 100μmol / L, 200μmol / L, 300μmol / L, 400μmol / L and 500μmol / L H2O2 respectively. (RT-PCR) to study the difference of Grx mRNA expression in each group. Results MTT results showed that 100μmol / L H2O2 had no effect on cell proliferation, and 200 ~ 500μmol / L H2O2 inhibited cell proliferation in varying degrees (P <0.05). The results of RT-PCR showed that the expression of Grx mRNA in ECV304 cells increased at the concentration of 100 μmol / LH 2 O 2 for 10 min, 30 min and 60 min, reaching the maximum at 30 min (P <0.05). The dose-dependent The results showed that the expression of Grx mRNA increased at the concentration of 100μmol / L H2O2 ~ 400μmol / L H2O2 for 30min, and the difference was significant (P <0.05) P <0.01). Conclusion H2O2 can affect the expression of Grx mRNA in human umbilical vein endothelial cells.