摘要
目的:观察珠子参总皂苷对H2O2诱导新生大鼠心肌细胞凋亡的抑制作用,并探讨其作用机制。方法:Wistar乳鼠心肌细胞原代培养,用H2O2建立氧化应激损伤模型,然后用珠子参总皂苷(100,200μg/mL)孵育24h后,MTT法检测珠子参总皂苷对细胞活力的影响,流式细胞仪和Hoechst33258染色检测珠子参总皂苷对氧化应激诱导细胞凋亡及胞内ROS含量的影响,比色法测定心肌细胞Caspase-3、Caspase-9的活性,荧光定量PCR测定心肌细胞Bcl-2和Bax mRNA表达,并计算Bcl-2与Bax的比值。结果:珠子参总皂苷(100,200μg/mL)能显著改善心肌细胞活性,有效保护线粒体膜电位的稳定,抑制心肌细胞凋亡和改善细胞形态,降低细胞内活性氧(ROS)含量;下调Bax mRNA表达,上调Bcl-2mRNA表达及Bcl-2与Bax的比值;降低H2O2所致新生大鼠心肌细胞中Caspase-3、Caspase-9的活性。结论:珠子参总皂苷对H2O2诱导心肌细胞凋亡有显著的抑制作用,其机制可能与其稳定心肌细胞膜、清除ROS及调节心肌细胞Bcl-2、Bax和Caspase-3、Caspase-9表达有关。
AIM： To observe the inhibition effects of total saponin from Panacis majoris rhizoma on myocardial apoptosis of neonatal rat in duced by H2O2 and its mechanism. METHODS： Oxidative stress-induced injury model was estab lished with H2O2. Effects of total saponin from Panacis majoris rhizome （100, 200μg/mL） on viability of cells and oxidative stress-induced apoptosis of cells were detected by MTT assay; Cell apoptosis rate and reactive oxygen species （ROS） content were detected by flow cytometry, the apoptosis cells morphous were observed by hoechst33258 through fluorescence microscope; The levels of Caspase-3, 9 were measured by detecting kits; Real-time polymerase chain reaction was applied to detect the mRNA expressions of Bcl-2 and Bax apoptosis-related genes. RESULTS.Total saponin from Panacis majoris rhizome （100, 200 μg/mL） might significantly increase cell viability and rnitoehondrial membrane potential, inhibit myocardial apoptosis, improve cell morphous, decrease intra-cellular ROS contents, down-regulate mRNA expression of Bax, up-regulate the mRNA expression of Bcl-2 and Bcl-2/Bax ratio, decrease activities of Caspase- 3, 9, and with the dose of total saponin increasing, the aforesaid improvement became more and more strong. CONCLUSION： Total saponin from Panacis majoris rhizome could effectively inhibit myocardial apoptosis of neonatal rat induced by H202. The mechanism may be related to stabilizing mitochondrial membrane potential, scavenging of ROS, regulating the Caspase-3, 9, Bcl-2 and Bax expression levels.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2012年第8期860-867,共8页
Chinese Journal of Clinical Pharmacology and Therapeutics
