[关键词]
[摘要]
本文研究了松多酚对H2O2致牛肺动脉内皮细胞(BPAECs)氧化应激损伤的保护作用及其相关机制。以H2O2100 μM诱导牛肺动脉内皮细胞(BPAECs)建立氧化损伤模型,实验设对照组、模型组、松多酚高、中、低(0.5、0.1、0.05 mg/mL)剂量组,免疫荧光检测转录因子NF-E 2相关因子2(Nrf2)核转位变化;RT-PCR检测Nrf2 mRNA的表达;Western blot检测Nrf2蛋白的表达;以及应用抑制剂筛选其发挥抗氧化损伤保护作用的信号通路。松多酚可以促进H2O2所致氧化损伤的牛肺动脉内皮细胞中的Nrf2从细胞质向细胞核内的转移,相对于模型组,松多酚提取物可以增加Nrf2 mRNA和蛋白质水平的表达量(*p<0.05,** p<0.01),且呈剂量-效应关系(** p<0.01)。松多酚对H2O2诱导氧化损伤的BPAECs具有保护作用,激活Nrf2/ARE机制可能与COX-2、p38、PI3K/AKT信号通路有关,而与PKC信号通路无关。
[Key word]
[Abstract]
The protective effect of polyphenols from Pinus sylvestris L. against hydrogen peroxide (H2O2)–induced oxidative stress in bovine pulmonary artery endothelial cells (BPAECs) was examined. An oxidative stress model was established by treating BPAECs with 100 μM H2O2. The BPAECs were divided into control group, H2O2 model group, and high-, medium-, and low-dose polyphenol groups (0.5, 0.1, and 0.05 mg/mL, respectively). Changes in the nuclear translocation of nuclear factor E2-related factor 2 (Nrf2) were detected by immunofluorescence, mRNA expression of Nrf2 was detected by reverse transcription-polymerase chain reaction (RT-PCR), and protein expression of Nrf2 was detected by western blotting. Various inhibitors were used to screen signaling pathways to determine the pathway that exerts the protective effect of polyphenols against oxidative stress. The polyphenols from P. sylvestris L. promoted the transfer of Nrf2 from the cytoplasm to the nucleus in BPAECs under hydrogen peroxide-induced oxidative stress. Compared with the control, the polyphenols from P. sylvestris L. significantly increased Nrf2 mRNA and protein levels (*p < 0.05, ** p < 0.01) in a dose-dependent manner (**p < 0.01). The polyphenol extract from P. sylvestris L. exerts a protective effect against hydrogen peroxide-induced damage in BPAECs, and likely activates Nrf2/ARE through COX-2, P38, and PI3K/AKT signaling pathways, but not the PKC pathway.
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[基金项目]
国家自然科学基金(31170510)