柴胡皂苷D对H2O2诱导大鼠心肌细胞H9c2损伤保护机制研究  

Protective effect of saikosaponin D on H2O2-induced injury in rat cardiomyocytes H9c2 cells

作　　者：丁雨[1] 陈雪梅[1] 吴思思[1] DING Yu;CHEN Xuemei;WU Sisi(Core Facility Center of West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院公共实验技术中心

出　　处：《浙江医学》2019年第24期2571-2575,2650,I0002,共7页Zhejiang Medical Journal

基　　金：国家自然科学基金资助项目（81870221)

摘　　要：目的探讨柴胡皂苷D对H2O2诱导大鼠心肌细胞H9c2损伤保护作用及机制。方法培养H9c2细胞,利用H2O2(400μM,4h)建立心肌细胞损伤模型。设立空白对照组、心肌损伤模型组(H2O2组)、柴胡皂苷D组(4μM),加入柴胡皂苷D预作用3h后,换成含浓度为400μM H2O2的完全培养基,继续培养4h后,应用多功能酶标仪检测细胞内活性氧水平,Hoechst 33258染色、流式细胞仪检测细胞凋亡变化,利用检测试剂盒检测细胞乳酸脱氢酶(LDH)和丙二醛(MDA)含量,同时利用荧光定量PCR(real-time PCR)技术检测过氧化物酶体增殖子活化受体(PPARγ)、核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)基因水平的表达变化。结果 Hoechst 33258染色和流式细胞检测结果显示:与对照组比较,H2O2组凋亡细胞升高,而柴胡皂苷D组的凋亡细胞量明显少于H2O2组(均P<0.05);细胞内活性氧(ROS)含量检测结果显示:H2O2组的细胞内ROS含量明显高于对照组,而柴胡皂苷D组明显低于H2O2组(均P<0.05);与对照组比较,H2O2组细胞LDH、MDA含量明显升高,柴胡皂苷D组细胞LDH、MDA含量明显降低(均P<0.05);柴胡皂苷D组的PPARγ、Nrf2、HO-1在m RNA水平的表达较H2O2组升高(均P<0.05)。结论柴胡皂苷D能够降低H2O2诱导H9c2心肌细胞产生的氧化应激反应,对心肌细胞具有保护作用。Objective To investigate the protective effect and mechanism of saikosaponin D(SD) on H2O2-induced injury in rat cardiomyocytes H9 c2 cells. Methods H9 c2 cells were treated with 400 μM H2O2 for 4 h to induce a myocardial cell injury model. H9 c2 cells in SD group were treated with 4μM saikosaponin D for 3 h before exposed to 400μM H2O2. The level of reactive oxygen species in the cells was detected by multi-function microplate reader, Hoechst 33258 staining and flow cytometry were used to detect cell apoptosis. The cell lactate dehydrogenase(LDH) and aldehyde(MDA) content were measured by the detection kit. Meanwhile, the expression of peroxisome proliferator-activated receptor gamma(PPARγ), nuclear factor E2-related factor 2(Nrf2), and heme oxygenase-1(HO-1) were detected by real-time RT-PCR. Results Hoechst 33258 staining and flow cytometry showed that the apoptotic cells in the H2O2 group increased compared with the control group, while the apoptotic cells in SD group were significantly decreased compared with the H2O2 group(all P<0.05).The intracellular ROS content in the H2O2 group was significantly higher than that in the control group, while that in SD group was significantly decreased compared with the H2O2 group(all P<0.05). Compared with the control group, the H2O2 group had significantly higher LDH release and cell MDA content, while those in SD group were significantly decreased(all P<0.05). The expression of PPARγ, Nrf2 and HO-1 in the SD group was higher than that in the H2O2 group(all P<0.05). Conclusion Saikosaponin D can reduce the oxidative stress induced by H2O2 in cardiomyocyte H9 c2 cells so that to protect the cell injury.

关 键 词：心肌细胞 柴胡皂苷 D 乳酸脱氢酶 丙二醛 过氧化物酶体增殖子活化受体 

分 类 号：R28[医药卫生—中药学]