Deng Huiyan, Li Huaping, Ho Zi Yin, Dai Xin Yue, Chen Quan, Li Runxiang, Liang Bihua, Zhu Huilan
Pharmazie. 2018 Nov 1;73(11):651-658. doi: 10.1691/ph.2018.8598.
The aim of this study was to further elucidate the mechanism of pterostilbene against UVA/UVB irradiation and the Nuclear factor E2-related factor 2 (Nrf2) signal pathway. A photo-damage model with UVA/UVB irradiation in HaCat cells was established and used in this study. The dose of pterostilbene was selected through MTS assay. Cell proliferation and apoptosis in Nrf2 and knockdown Nrf2 cells was detected by MTS assay. Expression of CAT, HO-1, and SOD in Nrf2 and knockdown Nrf2 cells was explored by qPCR. Western blot was used to analysis of Nrf2 nuclear translocation changes in Nrf2 and knockdown Nrf2 cells. Protein carbonyl content and MDA content was tested. : Our photo-damage model was successfully established and 20J/cm² UVA and 57mJ/cm² UVB irradiation was the suitable dose for HaCaT cell damage study. UVA/UVB irradiation would affect Nrf2 protein location, especial for 9.75 μM pterostilbene dose. In addition, cell proliferation could be significantly inhibited by UVA/UVB treatments (P<0.05), whereas, 9.75 μM pterostilbene treatment can alleviate the photo-damage. UVA/UVB irradiation would lead to decreased expressions of CAT, HO-1, and SOD. Carbonyl content and MDA was significantly changed by UVA/UVB treatments (P<0.05). The adverse events could be reversed by adding 9.75 μM pterostilbene. Western blot analysis showed that Nrf2 cytoplasm content in UVA/UVB treated cells was reduced and Nrf2 nuclear content was increased, which are different with the normal HaCaT cells without knockdown Nrf2 treatment (P<0.05). The results of cell proliferation, apoptosis, and cell antioxidant capacity in knockdown Nrf2 treated HaCaT cells were also significantly different with the normal HaCaT cells without knockdown Nrf2 treatment (P<0.05). We hypothesize that pterostilbene could play an anti-oxidation role the Nrf2 signal pathway.
本研究的目的是进一步阐明紫檀芪抗UVA/UVB辐射的机制以及核因子E2相关因子2（Nrf2）信号通路。本研究建立并使用了HaCaT细胞中UVA/UVB辐射的光损伤模型。通过MTS法选择紫檀芪的剂量。通过MTS法检测Nrf2和敲低Nrf2细胞中的细胞增殖和凋亡。通过qPCR探索Nrf2和敲低Nrf2细胞中CAT、HO-1和SOD的表达。使用蛋白质印迹法分析Nrf2和敲低Nrf2细胞中Nrf2核转位的变化。检测蛋白质羰基含量和丙二醛含量。：我们成功建立了光损伤模型，20J/cm² UVA和57mJ/cm² UVB辐射是用于HaCaT细胞损伤研究的合适剂量。UVA/UVB辐射会影响Nrf2蛋白的定位，特别是在9.75μM紫檀芪剂量下。此外，UVA/UVB处理可显著抑制细胞增殖（P<0.05），而9.75μM紫檀芪处理可减轻光损伤。UVA/UVB辐射会导致CAT、HO-1和SOD的表达降低。UVA/UVB处理显著改变了羰基含量和丙二醛含量（P<0.05）。添加9.75μM紫檀芪可逆转这些不良事件。蛋白质印迹分析表明，UVA/UVB处理的细胞中Nrf2细胞质含量降低，Nrf2核含量增加，这与未进行敲低Nrf2处理的正常HaCaT细胞不同（P<0.05）。敲低Nrf2处理的HaCaT细胞中的细胞增殖、凋亡和细胞抗氧化能力结果也与未进行敲低Nrf2处理的正常HaCaT细胞显著不同（P<0.05）。我们假设紫檀芪可能在Nrf2信号通路中发挥抗氧化作用。
