大鼠H9c2心肌细胞有氧氧化及无氧酵解的影响因素探讨

摘要

Objective This study was designed to explore the influence factors on H9 c2 cells energy metabolism with cellular energy metab-olism detection system (Seahorse XF Analyzer).Methods Different cell density of H9 c2 rat myocardial cell were cultured in SeahorseXF24 orifice plate, glycolysis was detected by glycolysis kits.With 105/mL cell density, different concentration FCCP stimulate cells, mito-chondrial function was detected using mitochondrial aerobic oxidation kits.Results The density of cells respond well to 2×104 cell numberof oligomycin stimulation rate value was better.50-400 range was the more appropriate cell density.2μmol/L FCCP could stimulate themaximum reaction, which was more appropriate concentration.Conclusion The cell density and FCCP concentration are the main factorsinfluencing the determination of cardiomyocytes glycolysis and mitochondrial respiratory.H9 c2 cardiomyocytes in 2×104/well cell inoculationdensity, and FCCP concentration 2μmol/L stimulate cells will be suitable for mitochondrial function test.%目的 利用细胞能量代谢检测系统(seahorse XF analyzer)探讨H9C2大鼠心肌细胞能量代谢的影响因素.方法 H9c2大鼠心肌细胞贴壁培养后以不同细胞密度接种于Seahorse XF24孔板,利用糖酵解压力试剂盒检测糖酵解压力曲线;以105/mL密度接种细胞,以不同浓度碳酰氰-4-三氟甲氧基苯胺(FCCP)刺激细胞,利用线粒体有氧氧化试剂盒检测线粒体功能.结果 各密度下细胞反应良好,以2×104细胞数对寡霉素刺激后的速率数值较好,为50～400范围,是较合适的细胞密度;2μmol/L FCCP能刺激最大反应,为合适的FCCP浓度.结论 细胞密度和FCCP浓度是影响心肌细胞有氧氧化及无氧酵解功能测定的主要因素,H9c2心肌细胞以2×104/孔细胞密度接种,FCCP浓度2μmol/L刺激细胞将适合细胞线粒体功能的检测.