ZHENG Feng,JIN Mei-hua,JIN Ming,et al.Inhibitory Effect of Soy Isoflavones on Apoptosis of L02 Cells Induced by Hydrogen Peroxide[J].Soybean Science,2020,39(04):621-625.[doi:10.11861/j.issn.1000-9841.2020.04.0621]
大豆异黄酮对过氧化氢致L02细胞凋亡的抑制作用
- Title:
- Inhibitory Effect of Soy Isoflavones on Apoptosis of L02 Cells Induced by Hydrogen Peroxide
- Keywords:
- Soybean; Isoflavones; L02; Hydrogen peroxide; Apoptosis
- 文献标志码:
- A
- 摘要:
- 为研究大豆异黄酮对过氧化氢所致L02细胞凋亡的抑制作用及其可能机制，以过氧化氢诱导L02细胞制备L02肝细胞凋亡模型。用Hoechst 33342荧光染色技术观察L02细胞核形态变化，用原位末端标记（TUNEL）技术观察L02细胞凋亡情况，采用免疫印迹法检测L02细胞中Caspase-3活性片段（Cleaved caspase-3）、B细胞淋巴瘤/白血病-2（Bcl-2）、Bcl-2相关X蛋白（Bax）、聚腺苷二磷酸核糖聚合酶活性片段（Cleaved PARP）表达，以及c-Jun氨基末端激酶（JNK）和核转录因子-κB（NF-κB）活化情况。结果显示：过氧化氢处理L02细胞能使细胞核Hoechst染色和TUNEL染色增强，提示L02细胞凋亡增多。同时，过氧化氢上调L02细胞Cleaved caspase-3、Cleaved PARP表达（P<0.05）、Bax/Bcl-2比值（P<0.05）和NF-κB p65核转位水平（P<0.05）。与损伤组比较，大豆异黄酮组L02细胞凋亡显著减少，Cleaved caspase-3和Cleaved PARP表达量、Bax/Bcl-2比值和NF-κB p65核转位水平均降低（P<0.05）。结果表明大豆异黄酮能抑制过氧化氢所致L02细胞凋亡，其作用可能与NF-κB通路有关。
- Abstract:
- In order to study the inhibitory effect of soybean isoflavone on apoptosis of L02 cells induced by hydrogen peroxide and its possible mechanism, we investigated the inhibitory effect of soy isoflavones on apoptosis of L02 cells induced by hydrogen peroxide. L02 apoptotic model was established by treating L02 cells with hydrogen peroxide. The morphological change of nucleus was detected by Hoechst 33342 fluorescent staining, cell apoptosis was detected by the TdT-mediated dUTP nick end labeling(TUNEL) method, the protein expression of the cleaved cysteinyl aspartate specific proteinase-3 (caspase-3), B cell lymphoma/lewkmia-2(Bcl-2), Bcl-2-associated x protein(Bax), cleaved poly(ADP-ribose) polymerase(PARP), as well as the activation of c-Jun N-terminal kinase (JNK) and nuclear factor-κB (NF-κB) were determined with the western blotting method. The results showed that hydrogen peroxide elevated both Hoechst fluorescent staining and TUNEL staining of L02 cells, indicative of the increased number of apoptotic cells induced by hydrogen peroxide. In addition, hydrogen peroxide up-regulated the expressions of cleaved caspase-3 and cleaved PARP (P<0.05), increased the ratio of Bax/Bcl-2 (P<0.05), and reduced the nuclear translocation of NF-κB p65 in L02 cells (P<0.05). However, soy isoflavones suppressed cell apoptosis, down-regulated the expressions of cleaved caspase-3 and cleaved PARP (P<0.05), reduced the Bax/Bcl-2 ratio (P<0.05), and decreased the nuclear translocation of NF-κB p65 (P<0.05) of L02 cells induced by hydrogen peroxide. It is suggested that soy isoflavones could suppress the apoptosis of L02 cells induced by hydrogen peroxide, probably through the regulation of NF-κB pathway.
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