益气活血中药参附益心颗粒对氧化损伤的心肌细胞活性的影响

The effects of Shenfu Yixin granule on the activity of oxidized cardiomyocytes by a long time dynamic living cell imaging system

目的研究参附益心颗粒(SFG)对H9C2心肌细胞增殖及H2O2氧化损伤的影响。方法利用IncuCyte长时间动态活细胞成像及功能分析系统IncuCyteS3,分别检测不同浓度参附益心颗粒(0.5、0.25、0.125、0.0625、0.03125mg/mL)对H9C2心肌细胞活性的影响;利用MTT法检测H2O2氧化损伤对心肌细胞活性的影响,并确定合适损伤浓度;心肌细胞在100μg/mL H2O2作用后,分别利用参附益心颗粒预孵育30min,再进行H2O2氧化损伤造模,IncuCyteS3系统检测其细胞增殖变化;同时加入荧光染料YOYO-1共培养,荧光显微镜拍照分析凋亡率。结果不同浓度参附益心颗粒处理心肌细胞不同的时间,未见明显细胞毒性;参附益心颗粒预孵育30min,再以50μm浓度H2O2刺激细胞,可不同程度的改善氧化损伤的程度;同时,参附益心颗粒预孵育一段时间后,与模型组比较,细胞凋亡率有所下降。结论参附益心颗粒可能能够改善氧化损伤的心肌细胞损伤,并改善由此引起的细胞凋亡。

Objective To study the effects of Shenfu Yixin Granules(SFG)on H9 C2 cardiomyocyte proliferation and H2O2 oxidative damage.Methods IncuCyte long-term dynamic live cell imaging and functional analysis system IncuCyteS3 was used to detect the effects of different concentrations of Shenfu Yixin granules(0.5,0.25,0.125,0.0625,0.03125 mg/mL)on the activity of H9 C2 cardiomyocytes;using MTT method,the effect of H2O2 oxidative damage on the activity of cardiomyocytes was detected,and the appropriate damage concentration was determined.After the treatment of 100μg/mL H2O2,the cardiomyocytes were pre-incubated with Shenfu Yixin granules for 30 min,and then H2O2 oxidative damage modeling was performed.IncuCyteS3 system was used for detection.The cell proliferation was changed;the fluorescent dye YOYO-1 was co-cultured,and the apoptosis rate was analyzed by fluorescence microscopy.Results Different concentrations of Shenfu Yixin granules treated cardiomyocytes for different time,no obvious cytotoxicity;Shenfu Yixin granules were pre-incubated for 30 min,then stimulated with 50μm H2O2,which could improve the degree of oxidative damage to varying degrees;After pre-incubation of Shenfu Yixin granules for a period of time,the apoptosis rate decreased compared with the model group.Conclusion The results of this experiment suggest that Shenfu Yixin granules may improve myocardial cell damage induced by oxidative damage and improve the resulting apoptosis.