大鼠H9c2心肌细胞有氧氧化及无氧酵解的影响因素探讨  

Research on the Influence Factors of H9c2 Cardiomyocytes Glycolysis and Mitochondrial Respiratory Function in Rats

作　　者：李彬[1] 崔琳[1] 王新陆[2] 王幼平[1] 谢世阳[1] 郝轩轩 高原[1] 王小晓[1] 朱明军[1] LI Bin;CUI Lin;WANG Xinlu;WANG Youping;XIE Shiyang;HAO Xuanxuan;GAO Yuan;WANG Xiaoxiao;ZHU Mingjun(First Affiliated Hospital,Henan University of Chinese Medicine,Zhengzhou 450000,Henan,China)

机构地区：[1]河南中医药大学第一附属医院,郑州450000 [2]河南中医药大学

出　　处：《中西医结合心脑血管病杂志》2019年第1期60-64,166-167,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：国家自然科学基金委项目(No.81503419;81373853;81703897)

摘　　要：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。Objective This study was designed to explore the influence factors on H9c2 cells energy metabolism with cellular energy metabolism detection system(Seahorse XF Analyzer).Methods Different cell density of H9c2 rat myocardial cell were cultured in Seahorse XF24 orifice plate,glycolysis was detected by glycolysis kits.With 105/mL cell density,different concentration FCCP stimulate cells,mitochondrial function was detected using mitochondrial aerobic oxidation kits.Results The density of cells respond well to 2×10^4 cell number of oligomycin stimulation rate value was better.50 400 range was the more appropriate cell density.2μmol/L FCCP could stimulate the maximum reaction,which was more appropriate concentration.Conclusion The cell density and FCCP concentration are the main factors influencing the determination of cardiomyocytes glycolysis and mitochondrial respiratory.H9c2 cardiomyocytes in 2×10^4/well cell inoculation density,and FCCP concentration 2μmol/L stimulate cells will be suitable for mitochondrial function test.

关 键 词：参附益心颗粒 心肌细胞 能量代谢 有氧氧化 无氧酵解 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]