小豆蔻明对H9c2细胞氧化损伤的保护作用

The protective effect of cardamonin on oxidative damage of H9c2 cells

目的 探讨小豆蔻明(cardamonin,CAR)对H2O2诱导的H9c2大鼠心肌细胞氧化损伤的保护作用及其初步作用机制.方法 提取山核桃叶中小豆蔻明,采用MTS(溴化噻唑蓝四氮唑)检测H2O2诱导氧化应激损伤的H9c2细胞存活率,Hoechst 33342染色观察其凋亡, Western blot技术检测p-AKT和AKT表达量,并用 PI3K/Akt 通路抑制剂LY294002 进行验证.结果 小豆蔻明在浓度为10μM时,能极显著提高H2O2诱导的H9c2细胞的存活率,心肌细胞存活率为(90.48±1.45)%;小豆蔻明能极显著的降低乳酸脱氢酶(LDH)活性和细胞丙二醛(MDA)含量,升高超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)和过氧化氢酶(CAT)的活力;小豆蔻明组能减轻细胞核凝聚固缩的现象,减少细胞碎片;明显增加p-Akt的表达,但加入LY294002后,p-Akt蛋白的表达明显被抑制.结论 小豆蔻明对H2O2诱导的H9c2氧化应激具有一定的保护作用,其机制可能通过PI3K/Akt信号通路抑制H2O2诱导的H9c2细胞凋亡,与降低脂质过氧化效应,提高内源性抗氧化酶的活性有关.

Objective To study the protective effect of cardamonin(CAR)on oxidative damage of H9c2 from leaf of Carya cathayensis against H2O2-induced oxidative injury in H9c2 myocardial cells and its relationship with PI3K/Akt signaling pathway.Methods Extraction of cardamom from C.cathayensis.The cell viability of H2O2-induced oxidative stress injury cells was measured by MTS,their apoptosis was determined by Hoechst 33342 staining,and their expressions of p-AKT and AKT were measured by Western blot.The effects were verified by using PI3K/Akt inhibitor LY294002.Results Cardamonin significantly increased the survival rate of H9c2 cells induced by H2O2,and the survival rate of cardiomyocytes was 90.48±1.45%.It significantly reduced lactate dehydrogenase(LDH)activity and cell malondialdehyde(MDA)content,and increased superoxide dismutase(SOD),glutathione peroxidase(GSH-PX)and The activity of catalase(CAT).Cardamonin significantly improved the cell viability of oxidative stress injury cell model,reduced the apoptosis of cells,and increased the expression of p-AKT,but LY294002 offsets such an effect.Conclusion Our findings suggest that cardamonin tended to improve the H9c2 myocardial cellular morphology,enhance cell activity and reduce apoptosis rate.Its mechanism of action in reducing oxidative stress injury may be related to the activation of PI3K/Akt signaling pathway.