摘要
目的探讨不同浓度槲皮素对H_2O_2诱导状态下H9C2心肌细胞氧化应激损伤的保护作用及作用机制。方法 H9C2心肌细胞传代培养,分为4组:对照组:以正常培养液培养;氧化损伤组:100μmol·L^(-1)的H_2O_2作用12 h;槲皮素低浓度组:10μmol·L^(-1)槲皮素孵育24 h后,加入H_2O_2作用12 h;槲皮素高浓度组:100μmol·L^(-1)槲皮素孵育24 h后,加入H_2O_2作用12 h。应用MTT法观察槲皮素对H9C2心肌细胞的生长抑制作用,流式细胞技术观查各组H9C2心肌细胞凋亡情况,H2DCFDA染色观察细胞内活性氧生成量,比色法检测凋亡相关因子Caspses-3及Caspase-9的表达。结果槲皮素明显抑制了H_2O_2诱导的H9C2心肌细胞活力的下降,降低细胞凋亡率,抑制细胞内ROS生成,明显减少H9C2心肌细胞内Caspses-3及Caspase-9的表达,槲皮素低、高浓度组与氧化损伤组比较,差异均有统计学意义(P<0.05)。结论槲皮素通过抑制Caspase-3及Caspase-9表达抑制H_2O_2诱导的H9C2心肌细胞凋亡,可能在心肌细胞损伤的治疗中具有一定作用。
Objective To explore protective effect of different concentrations of quercetin on H2O2 induced H9C2 cardiomyocytes oxidative stress damage and its mechanism. Methods H9C2 cardiomyoeytes were subcuhured and divided into four groups： control group： cultured in normal cultivation; oxidative damage group （treated with 100 μmol·L-1 H2O2 for 12 h）, quereetin low concentration group （10μmol·L-1 quereetin treated for 24 h and H2O2 treated for 12 h）, quercetin high concentration group （ 100μmol·L-1 quercetin treated for 24 h and H2O2 treated for 12 h）. MTT method was applied to observe the growth inhibition of quereetin on H9C2 cardiomyoeytes, flow cytometry technique was used to observe H9C2 cardiomyocytes apoptosis, H2DCFDA staining was used to observe intraeellular reactive oxygen species genera- tion, and eolorimetric method was used to detect the expression of apoptosis related factor Caspses-3 and Caspase-9. Results Quercetin obviously inhibited H2O2 -induced H9C2 cardiomyocytes vitality decline, decreased the rate of apoptosis, inhibited ROS generation in cells,significantly reduced the expression of Caspses-3 and Caspase-9 in H9C2 cardiomyocytes, when compared with oxidative damage group, the difference in the quercetin low, high concentration group had statistical significance （P 〈 0. 05）. Conclusion Quercetin inhibits H9C2 myocardial apoptosis by inhibiting the, expression of Caspses-3 and Caspase-9 in H9C2 cardiomyo- cytes, which indicate that quercetin may play a role in the treatment of myocardial cell damage.
出处
《哈尔滨医科大学学报》
CAS
2017年第5期387-390,共4页
Journal of Harbin Medical University
基金
哈尔滨科技创新人才研究基金项目(2014RFXGJ018)
