参附注射液对缺氧心肌细胞凋亡的抑制效应

Inhibiting effect of ginseng radix aconiti pareparata injection on hypoxia-induced cardiomyocyte apoptosis

目的 探讨参附注射液对缺氧诱导的培养乳鼠心肌细胞凋亡的作用。方法 将 SD乳鼠的培养心肌细胞随机分为三组:对照组、缺氧72 h组和参附注射液组。缺氧 72 h前加入1 mmol参附注射液。应用TUNEL法及流式细胞仪分析心肌细胞凋亡的变化,借助半胱胺酸 天门冬胺酸酶(caspase- 3)荧光分析试剂盒,荧光比色法检测心肌细胞凋亡过程中 caspase -3 活性的变化。结果 TUNEL及流式细胞仪分析显示,参附注射液组 AI值和细胞凋亡百分率分别为(14. 10±2 .56)%和(14. 93±2 .47)%,明显低于缺氧72 h组(46 .49±4 .93)%和(48 .43±4 .18)%(P<0 .01)。caspase -3活性检测显示,参附注射液组的caspase- 3活性比缺氧72 h组降低了 52. 85%(P<0. 01)。结论 参附注射液能明显抑制缺氧诱导的心肌细胞凋亡。caspase- 3蛋白酶活性抑制可能是其机理之一。

Objective To investigate the effect of ginseng radix aconiti praeparata injection on neonatal cardiomyocyte apoptosis induced by hypoxia. Methods The cultured cardiomyocytes from 2-3d newborn SD rats were randomly divided into 3 group: control group, ischaemia 72h group and ginseng radix aconiti praeparata injection group (addition of ginseng radix aconiti praeparata injection at 72h before ischaemia 72h). Cardiomyocyte apoptosis was determined by TUNEL and flow cytometry. The caspase-3 activity was detected by caspase-3 fluorescent assay kit . Results TUNEL methlod and flow cytometry analysis showed that apoptosis indexes and the percentage of apoptosis cells in ginseng radix aconiti praeparata injection group were (14.10±2.56)% and (14.93±2.47)% respectively, which were significantly lower than those in ischaemia 72h group [(46.49±4.93)%and(48.43±4.18)%](P<0.01). Caspase-3 activity analysis showed that caspase-3 activity in ginseng radix aconiti praeparata injection group was reduced by 52.85% relative to ischaemia 72h group. Conclusion Ginseng radix aconiti praeparata injection can efficiently inhibit apoptosis of neonatal cardiomyocytes after hypoxia. The inhibition of caspase-3 activity may be one of the mechanisms for that.