论文部分内容阅读
研究Sirt1蛋白对间充质干细胞(MSCs)受照射后产生的炎症因子IL-1β的影响,探讨Sirt1蛋白在辐射防护中的作用。shRNA体外沉默间充质干细胞Sirt1基因转录,检测细胞内Sirt1 mRNA表达证实基因沉默效果;同时给予间充质干细胞200μmol·L~(-1)白藜芦醇,经4 Gy照射后,用ELISA、Western blot和RT-PCR等方法检测Sirt1沉默对IL-1β分泌和表达的影响。结果表明,照射前沉默Sirt1基因并给予间充质干细胞200μmol·L~(-1)白藜芦醇,与照射前单纯给予200μmol·L~(-1)白藜芦醇相比,辐射引起的IL-1β分泌显著升高(t=18.57,p<0.05),同时细胞内IL-1β蛋白表达和mRNA水平也显著升高(t=8.078,p<0.05)。沉默Sirt1后可明显抑制白藜芦醇的辐射保护作用,提示Sirt1蛋白在辐射后产生炎症的过程中起到关键的调节作用。
To investigate the effect of Sirt1 on the inflammatory cytokine IL-1β produced by irradiation of mesenchymal stem cells (MSCs), and to explore the role of Sirt1 in radiation protection. shRNA silencing the Sirt1 gene transcription of mesenchymal stem cells in vitro and detecting the Sirt1 mRNA expression in the cells confirmed the gene silencing effect. At the same time, the mesenchymal stem cells were treated with 200 μmol·L -1 resveratrol. After 4 Gy irradiation, The effects of Sirt1 silencing on the secretion and expression of IL-1β were detected by Western blot and RT-PCR. The results showed that Sirt1 gene silencing and resveratrol with 200μmol·L -1 were better than those with 200μmol·L -1 resveratrol before irradiation The secretion of IL-1β was significantly increased (t = 18.57, p <0.05), and the level of IL-1β protein and mRNA was significantly increased (t = 8.078, p <0.05). The silence of Sirt1 significantly inhibited the radiation protection of resveratrol, suggesting that Sirt1 protein plays a key regulatory role in the process of inflammation after radiation.