维生素C对体外培养的H_9C_2心肌细胞系的影响

Effect of vitamin C on cultured H_9C_2 myocardial cells

目的:探讨维生素C对大鼠H_9C_2心肌细胞抗氧化能力与细胞活力的影响。方法:以体外培养的大鼠H_9C_2心肌细胞为研究对象,在培养过程中,分别加入不同浓度的维生素C,观察细胞形态,MTT法检测心肌细胞活力,DCFH-DA荧光探针法测定维生素C对H_2O_2处理的心肌细胞活性氧(ROS)生成的影响,用比色法检测半胱天冬酶(caspase)-3和caspase-9活性。结果:0.05 mg/mL维生素C处理后,H_9C_2心肌细胞形态发生变化,细胞质中颗粒状物质增多,排列紊乱,间隙增宽;0.05 mg/mL维生素C可有效抑制H_2O_2导致的ROS积累;但随着培养基中维生素C浓度的升高,心肌细胞存活率明显降低,且维生素C处理后caspase-3和caspase-9活性明显升高。结论:H_9C_2心肌细胞培养中添加维生素C可产生明显的抗氧化作用,但维生素C对H_9C_2心肌细胞生长具有抑制作用。

Objective：To investigate the effect of vitamin C on the antioxidant capacity and cell viability of H9C2 cell lines. Methods：The cultured H9C2 cells were treated with vitamin C of different concentrations, and cell morphology was observed under inverted microscope MTF assay was used to test the cell viability. Fluorescent probe DCFH-DA was used to determinate the effect of vitamin C on the intracellular ROS induced by H202. The activity of caspase-3 and caspase-9 were tested by eolorimetric method. Results： The morphological changes of Hgca cardiomyocyteswere observed after treatment of 0. 05 mg/mL vitamin C, including the increase of granular substance in the cytoplasm, arranged disorder and widened intercellular gap. 0. 05 mg/mL vitamin C could inhibit the accumulation of ROS caused by H202, but with the increase of vitamin C concentration, the survival rate of cardiomyocytes decreased significantly and the activity of caspase-3 and caspase-9 increased significantly. Conclusion： Vitamin C has a significant antioxidant effect on the H9 Ca cardiomyocytes, but it can significantly inhibit the cell growth.