H_2O_2对ECV304细胞Grx mRNA表达的影响(英文)

Effect of H_2O_2on Grx mRNAlevel in ECV304

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/L H2O2作用10min、30min、60min均能使ECV304细胞中Grx mRNA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 study the effect of different concentration of H2O2 on Grx mRNA level in human umbilical endothelium cells （ECV304）. Methods ECV304 cells were cultured in vitro. The cell proliferation was measured by MTT assay. In time-relative observation, cells were divided into six groups. Except control group, each other group was treated with 100μmol/L H2O2 for 5min, 10min, 30min, 60min, 120min, respectively; In dose-relative observation, cells were also divided into six groups. Except control group, each other group was treated with 100μmaol/L, 200μmol/L, 300μmol/L,400μmol/L and 500μmol/L H2O2 for 30min. The total RNA was extracted by Trizol one-step method in cells. Grx mRNA level in ＇each group was detected by means of RTPCR （Reverse Transcription Polymerase Chain Reaction）. Results MTT assay showed that the cell viability was decreased when the ceils were treated by 200 - 500μmol/L H2O2 （ P 〈 0.05） ; In time-relative observation, Grx mRNA level was increased when the ceils were given 100μmol/L H2O2 for 10,30,60minutes（ P 〈 0.05）; In dose-relative observation, Grx mRNA level was also increased when the cells were exposed to 100 - 400μmol/L H2O2 （ P 〈 0.05）. Moreover, Grx mRNA level reached its own highest point at 300μmol/L H2O2. Conclusion H2O2 affects Grx mRNA level in ECV304 cells.