Yang Kun, Wu Yue, Zhang Rong, Lei Xiao-Ping, Kang Lan, Dong Wen-Bin
Department of Neonatology, Children's Medical Center, Affiliated Hospital of Southwest Medical University/Sichuan Clinical Research Center for Birth Defects, Luzhou, Sichuan 646000, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2024 Aug 15;26(8):852-860. doi: 10.7499/j.issn.1008-8830.2404120.
To investigate the effect of reactive oxygen species (ROS)/silent information regulator 1 (SIRT1) on hyperoxia-induced mitochondrial injury in BEAS-2B cells.
The experiment was divided into three parts. In the first part, cells were divided into H0, H6, H12, H24, and H48 groups. In the second part, cells were divided into control group, H48 group, H48 hyperoxia+SIRT1 inhibitor group (H48+EX 527 group), and H48 hyperoxia+SIRT1 agonist group (H48+SRT1720 group). In the third part, cells were divided into control group, 48-hour hyperoxia+N-acetylcysteine group (H48+NAC group), and H48 group. The ROS kit was used to measure the level of ROS. Western blot and immunofluorescent staining were used to measure the expression levels of SIRT1 and mitochondria-related proteins. Transmission electron microscopy was used to observe the morphology of mitochondria.
Compared with the H0 group, the H6, H12, H24, and H48 groups had a significantly increased fluorescence intensity of ROS (<0.05), the H48 group had significant reductions in the expression levels of SIRT1 protein and mitochondria-related proteins (<0.05), and the H24 and H48 groups had a significant reduction in the fluorescence intensity of mitochondria-related proteins (<0.05). Compared with the H48 group, the H48+SRT1720 group had significant increases in the expression levels of mitochondria-related proteins and the mitochondrial aspect ratio (<0.05), and the H48+EX 527 group had a significant reduction in the mitochondrial area (<0.05). Compared with the H48 group, the H48+NAC group had a significantly decreased fluorescence intensity of ROS (<0.05) and significantly increased levels of SIRT1 protein, mitochondria-related proteins, mitochondrial area, and mitochondrial aspect ratio (<0.05).
The ROS/SIRT1 axis is involved in hyperoxia-induced mitochondrial injury in BEAS-2B cells.
探讨活性氧（ROS）/沉默信息调节因子1（SIRT1）对高氧诱导的BEAS - 2B细胞线粒体损伤的影响。
实验分为三个部分。第一部分，细胞分为H0、H6、H12、H24和H48组。第二部分，细胞分为对照组、H48组、H48高氧+SIRT1抑制剂组（H48 + EX 527组）和H48高氧+SIRT1激动剂组（H48 + SRT1720组）。第三部分，细胞分为对照组、48小时高氧+N - 乙酰半胱氨酸组（H48 + NAC组）和H48组。使用ROS试剂盒测量ROS水平。采用蛋白质免疫印迹法和免疫荧光染色法测量SIRT1和线粒体相关蛋白的表达水平。透射电子显微镜用于观察线粒体的形态。
与H0组相比，H6、H12、H24和H48组的ROS荧光强度显著增加（<0.05），H48组的SIRT1蛋白和线粒体相关蛋白的表达水平显著降低（<0.05），H24和H48组的线粒体相关蛋白荧光强度显著降低（<0.05）。与H48组相比，H48 + SRT1720组的线粒体相关蛋白表达水平和线粒体纵横比显著增加（<0.05），H48 + EX 527组的线粒体面积显著减小（<0.05）。与H48组相比，H48 + NAC组的ROS荧光强度显著降低（<0.05），SIRT1蛋白、线粒体相关蛋白水平、线粒体面积和线粒体纵横比显著增加（<0.05）。
ROS/SIRT1轴参与了高氧诱导的BEAS - 2B细胞线粒体损伤。
