大蒜素抑制H_2O_2诱导的心肌细胞凋亡的实验研究  

Effect of allicin on hydrogen peroxide-induced apoptosis in H9c2 cardiacmyocytes

作　　者：李辉[1] 赵锴[1] 王磊[1] 

机构地区：[1]河南省商丘市第一人民医院心血管内科二病区,河南商丘476000

出　　处：《现代预防医学》2016年第6期1097-1101,共5页Modern Preventive Medicine

摘　　要：目的探究大蒜素对过氧化氢(Hydrogen Peroxide,H_2O_2)诱导的H9c2细胞凋亡的影响。方法 H_2O_2和不同浓度大蒜素分别作用于H9c2细胞,Western blot检测对Caspase-3表达的影响,流式细胞术检测对细胞凋亡的影响,RT-PCR和Western blot检测对血红素氧合酶1(Heme Oxygenase-1,HO-1)表达的影响。分别通过HO-1抑制剂Zn PP9(Zinc protoporphyrin IX)和PI3K/AKT抑制剂LY294002,MTT检测对细胞增殖活性的影响,Western blot检测对p-Akt、核转录因子Nrf2(nuclear factor-erythroid 2 related factor 2)和HO-1等表达的影响。结果对照组和100μM H_2O_2+大蒜素(0、5、10、20、40 mg/L)组细胞凋亡率分别为(4.83±0.66)%、(68.38±6.97)%、(61.19±6.01)%、(47.32±5.21)%、(22.07±3.73)%和(20.05±2.93)%、与Caspase-3蛋白表达趋势一致,并具有浓度依赖性(P<0.05)。大蒜素作用下,HO-1 mRNA和蛋白表达亦呈浓度依赖性增加(P<0.05)。H_2O_2组细胞相对增殖活性降低至(61.38±1.76)%(P<0.05),大蒜素处理使其上调至(81.64±3.79)%(P<0.05),而Zn PP9可阻断该上调,为(69.82±2.22)%(P<0.05)。大蒜素明显增加p-Akt、Nrf2和HO-1等表达,可被LY294002阻断(P<0.05)。结论大蒜素可通过Akt/Nrf2途径上调HO-1表达并抑制H_2O_2诱导的H9c2细胞凋亡。Objective This study was to explore the effect of allicin on hydrogen peroxide（ H2O2）- induced apoptosis in H9c2 cardiacmyocytes. Methods Cultured cells were treated with H2O2 and allicin in different concentrations. The Caspase- 3expression was detected by western blot and the cell apoptosis rate was determined by flow cytometry,respectively. RT- PCR and western blot were applied to quantify the expression of heme oxygenase- 1（ HO- 1）. HO- 1 inhibitor Zn PP9 was introduced to determine the cell viability by MTT assay. PI3 K / AKT inhibitor LY294002 was introduced to determine the expression of p- Akt,nuclear factor- erythroid 2 related factor 2（ Nrf2） and HO- 1 proteins by western blot. Results The apoptosis rate of the control group and the groups with 100μM H2O2＋ allicin（ 0,5,10,20,and 40 mg / L） were（ 4. 83 ±0. 66） %,（ 68. 38 ± 6. 97） %,（ 61. 19 ± 6. 01） %,（ 47. 32 ± 5. 21） %,（ 22. 07 ± 3. 73） %,and（ 20. 05 ± 2. 93） %,respectively. These results were consistent with the dose- dependent Caspase- 3 expression（ P〈0. 05）. H2O2 reduced the cell proliferation activity to（ 61. 38 ± 1. 76） %（ P〈0. 05）,but allicin pretreatment up- regulated it to（ 81. 64 ± 3. 79） %（ P〈0. 05）. Such upregulation was blocked by Zn PP9,with the activity of（ 69. 82 ± 2. 22） %（ P〈0. 05）. In addition,allicin markedly elevated the expression of p- Akt,Nrf2 and HO- 1 proteins,which was notably attenuated by LY294002pretreatment（ P〈0. 05）. Conclusion Allicin may up- regulate HO- 1 expression and inhibit H2O2- induced cell apoptosis through Akt / Nrf2 pathway in H9c2 cardiacmyocytes.

关 键 词：大蒜素 心肌细胞 H2O2 血红素氧合酶-1 细胞凋亡 

分 类 号：R541[医药卫生—心血管疾病]