摘要
目的:探讨参芎葡萄糖注射液(Shenxiong glucose injection,SGI)对过氧化氢(H2O2)诱导的H9c2细胞氧化损伤的保护作用及其机制。方法:体外培养H9c2心肌细胞,用H2O2(300μmol·L-1)处理0.5 h,建立H9c2心肌细胞H2O2氧化损伤模型,SGI组:H9c2细胞用(100,200,400μmol·L-1)SGI处理6 h后,另设空白组与模型组,再用H2O2处理0.5 h。利用流式细胞术检测细胞内活性氧(ROS)和线粒体膜电位(MMP);用MTS法检测细胞存活率;用酶联免疫吸附测定(ELISA)法检测乳酸脱氢酶(LDH)漏出量、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性;用蛋白质免疫印迹(Western blot)技术检测凋亡相关基因B细胞淋巴瘤/白血病-2(Bcl-2),Bcl-2相关X蛋白(Bax)和半胱氨酸蛋白酶(Caspase-3)的表达情况。结果:在300μmol·L-1H2O2作用细胞0.5 h的情况下,细胞存活率降低至50%左右,降低的程度合适,且实验结果重复性好,因此后续实验用该条件建立氧化损伤模型。与模型组比较,SGI预处理6 h能显著升高细胞存活率(P<0.05,P<0.01),减少LDH的外漏和MDA的生成(P<0.05,P<0.01),显著增加SOD,GSH-Px和CAT的活性(P<0.01),明显减少H9c2细胞内ROS含量(P<0.01),并使MMP丢失减少(P<0.01),Western blot表明,SGI能显著上调Bcl-2的表达,下调Caspase-3的表达(P<0.05,P<0.01)。结论:参芎葡萄糖注射液能保护H9c2心肌细胞对抗H2O2诱导的氧化损伤,其作用机制可能与其提高细胞清除ROS能力和抑制细胞凋亡有关。
Objective： To investigate the protective effect of Shenxiong Glucose Injection（ SGI） on H2O2-induced oxidative damage of H9c2 cells. Method： The in vitro oxidative stress damage model was established by treating H9c2 cells with H2O2（ 300 μmol·m L- 1） for 0. 5 h. SGI-pretreated group： pretreated H9c2 cells with SGI（ 100,200,400 μmol·m L- 1） for 6 h. Blank group and model group were treated additionally with H2O2 for 0. 5 h. The intracellular reactive oxygen species（ ROS） and mitochondrial membrane potential（ MMP） were detected by flow cytometry. The cell viability was detected by MTS assay; ELISA methods was used to determine LDH release,MDA content,SOD,CAT,GSH-PX activity. The expression of apoptosis-related genes Caspase-3,Bax and Bcl-2 was detected by western blotting technology. Result： After H9C2 cells was treated with H2O2（ 300 μmol·m L- 1） for 0. 5 h,the survival rate of the cells decreased to about 50%. The experimental results were so appropriate and repeatable that the subsequent oxidative damage model was established under the conditions. Compared with the control group,the cells survival was significantly increased（ P 〈 0. 05,P 〈 0. 01）by pretreating with SGI for 6 h. SGI not only decreased the release of LDH（ P 〈 0. 05,P 〈 0. 01）,MDA（ P 〈0. 05,P 〈 0. 01）,but also increased the activity of SOD,GSH-PX,CAT（ P 〈 0. 01）. In addition,the content of ROS significantly decreased（ P 〈 0. 01）,and MMP was increased（ P 〈 0. 01）. Western blot showed SGI can significantly enhance the expression of Bcl-2 and weaken the expression of Caspase-3（ P 〈 0. 05,P 〈 0. 01）.Conclusion： SGI shows the protective effect on H2O2-induced oxidative damage of H9c2 cells. Its mechanism may be correlated with increase in ROS removal ability and inhibition of apoptosis.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2016年第8期153-158,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
贵阳市科技局重大专项([2011401]社6-2号)
贵州省科技厅重大专项(黔科合重大专项字[2011]6019号)
