CAJ | 학술논문

目的：采用过氧化氢（H2O2）诱导大鼠主动脉内皮细胞（SVAREC）氧化损伤，建立SVAREC氧化损伤模型。方法体外培养SVAREC细胞，绘制细胞生长曲线，用不同浓度的H2O2刺激细胞，并在不同时间点用MTS法测定细胞存活率，以确定SVAREC氧化损伤模型条件。倒置显微镜下观察细胞形态学变化，微孔酶标法测乳酸脱氢酶（LDH）活力，流式细胞术检测细胞凋亡率，WB检测细胞中半胱氨酸蛋白酶（Caspase-3）的表达情况。结果与空白对照组细胞比较，当H2O2浓度为100μM，作用时间为2 h时，造成细胞损伤，其细胞凋亡率和Caspase-3蛋白表达量明显高于空白对照组。结论该实验条件下H2O2浓度为100μmol/L，作用时间为2 h是模型的最佳复制条件。
목적：채용과양화경（H2O2）유도대서주동맥내피세포（SVAREC）양화손상，건립SVAREC양화손상모형。방법체외배양SVAREC세포，회제세포생장곡선，용불동농도적H2O2자격세포，병재불동시간점용MTS법측정세포존활솔，이학정SVAREC양화손상모형조건。도치현미경하관찰세포형태학변화，미공매표법측유산탈경매（LDH）활력，류식세포술검측세포조망솔，WB검측세포중반광안산단백매（Caspase-3）적표체정황。결과여공백대조조세포비교，당H2O2농도위100μM，작용시간위2 h시，조성세포손상，기세포조망솔화Caspase-3단백표체량명현고우공백대조조。결론해실험조건하H2O2농도위100μmol/L，작용시간위2 h시모형적최가복제조건。
Objective To establish the oxidative damage model of SVAREC. Methods SVAREC were cultured in vitro and the growth curves were draw. Different concentration of H2O2 were stimulated the cells,and the cell viability were measured by the method of MTS at different time. Under the condition of this model,cellular morphology were observed by inverted microscope,the activity of LDH were measured by microplate reader,the apoptosis rate of cells also were detected by flow cytometry,and Caspase-3 expression were tested by Western blotting. Results Compared with control,cells were injured at the concentration of H2O2 is 100μmol/L for 2 h,which,the apoptosis rate and protein expression of Caspase-3 were significantly higher. Conclusion The best replication model of this experimental condition is exposure the SVAREC at the 100μmol/L concentration of H2O2 for 2 h.