Department of Dermatology, Xiangya Hospital, Central South University, Changsha, China.
Department of Pediatrics, Xiangya Hospital, Central South University, Changsha, China.
J Dermatol. 2018 Jul;45(7):791-798. doi: 10.1111/1346-8138.14304. Epub 2018 May 3.
The aim of this study was to investigate the protective effect of fluorofenidone (5-methyl-1-[3-fluorophenyl]-2-[1H]-pyridone, AKF-PD) on ultraviolet (UV)-A-induced senescence in human dermal fibroblasts (HDF) and examine the mechanisms involved. HDF were treated with AKF-PD. Senescence-associated (SA)-β-galactosidase level, cell viability and expression of p16 were evaluated. In addition, UV-A-irradiated HDF were treated with AKF-PD, rapamycin and MHY1485; SA-β-galactosidase staining, 3-(4 5-dimethylthiazol-2-yl)-2 5-diphenyltetrazolium bromide assay and western blot for SIRT1 were performed; and phosphorylated mammalian target of rapamycin (p-mTOR) expression and reactive oxygen species (ROS) levels were measured. Intracellular ROS was detected by the 2',7'-dichlorofluroescein diacetate probe. Our results showed that AKF-PD substantially attenuated the changes of p16 expression, SA-β-galactosidase staining and cellular proliferation induced by UV-A irradiation in HDF. AKF-PD rescued the increased mTOR phosphorylation and reduced SIRT1 expression induced by UV-A irradiation in HDF. AKF-PD and rapamycin together had a synergistic effect on p-mTOR reduction and SIRT1 increase. mTOR activator MHY1485 partly blocked the above effects. Moreover, intracellular ROS level induced by UV-A irradiation could partly decrease by AKF-PD, and MHY1485 could reduce this effect. Our results indicated that AKF-PD could alleviate HDF senescence induced by UV-A-irradiation by inhibiting the p-mTOR and increasing SIRT1. Moreover, AKF-PD may be a potential treatment material for skin.
本研究旨在探讨氟芬酮（5-甲基-1-[3-氟苯基]-2-[1H]-吡啶酮，AKF-PD）对人真皮成纤维细胞（HDF）紫外线（UV）-A 诱导衰老的保护作用，并探讨其相关机制。用 AKF-PD 处理 HDF。评估衰老相关（SA）-β-半乳糖苷酶水平、细胞活力和 p16 的表达。此外，用 AKF-PD、雷帕霉素和 MHY1485 处理经 UV-A 照射的 HDF；进行 SA-β-半乳糖苷酶染色、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐测定和 SIRT1 的 Western blot；并测量磷酸化哺乳动物雷帕霉素靶蛋白（p-mTOR）表达和活性氧（ROS）水平。通过 2'，7'-二氯二氟荧光素二乙酸探针检测细胞内 ROS。结果显示，AKF-PD 显著减弱了 UV-A 照射诱导的 HDF 中 p16 表达、SA-β-半乳糖苷酶染色和细胞增殖的变化。AKF-PD 挽救了 UV-A 照射诱导的 HDF 中 mTOR 磷酸化增加和 SIRT1 表达降低。AKF-PD 和雷帕霉素联合使用对 p-mTOR 减少和 SIRT1 增加具有协同作用。mTOR 激活剂 MHY1485 部分阻断了上述作用。此外，UV-A 照射诱导的细胞内 ROS 水平可部分被 AKF-PD 降低，而 MHY1485 可降低此作用。结果表明，AKF-PD 通过抑制 p-mTOR 和增加 SIRT1 可减轻 UV-A 照射诱导的 HDF 衰老。此外，AKF-PD 可能是一种有潜力的皮肤治疗药物。
