Zhang Chunli, Yuchi Haishen, Sun Lu, Zhou Xiaoli, Lin Jinde
Department of Clinical Research, Friendship Plastic Surgery Hospital, Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China.
Mol Med Rep. 2017 Aug;16(2):2016-2022. doi: 10.3892/mmr.2017.6795. Epub 2017 Jun 15.
The aim of the present study was to determine if human amnion‑derived mesenchymal stem cells (HAMSCs) exert a protective effect on ultraviolet A (UVA) irradiation-induced human dermal fibroblast (HDF) senescence. A senescence model was constructed as follows: HDFs (104‑106 cells/well) were cultured in a six‑well plate in vitro and then exposed to UVA irradiation at 9 J/cm2 for 30 min. Following the irradiation period, HDFs were co‑cultured with HAMSCs, which were seeded on transwells. A total of 72 h following the co‑culturing, senescence‑associated β‑galactosidase staining was performed and reactive oxygen species (ROS) content and mitochondrial membrane potential (Δψm) were detected in the HDFs via flow cytometric analysis. The results demonstrated that the percentage of HDFs, detected via staining with X‑gal, were markedly decreased when co‑cultured with human HAMSCs, compared with the group that were not co‑cultured. The ROS content was decreased and the mitochondrial membrane potential (Δψm) recovered in cells treated with UVA and HAMSCs, compared with that of cells treated with UVA alone. Reverse transcription‑quantitative polymerase chain reaction revealed the significant effects of HAMSCs on the HDF senescence marker genes p53 and matrix metalloproteinase‑1 mRNA expression. In addition to this, western blot analysis verified the effects of HAMSCs on UVA induced senescence, providing a foundation for novel regenerative therapeutic methods. Furthermore, the results suggested that activation of the extracellular‑signal regulated kinase 1/2 mitogen activated protein kinase signal transduction pathway, is essential for the HAMSC‑mediated UVA protective effects. The decrease in ROS content additionally indicated that HAMSCs may exhibit the potential to treat oxidative stress‑mediated UVA skin senescence in the future.
本研究的目的是确定人羊膜间充质干细胞（HAMSCs）是否对紫外线A（UVA）照射诱导的人皮肤成纤维细胞（HDF）衰老具有保护作用。衰老模型构建如下：将HDFs（104-106个细胞/孔）在体外六孔板中培养，然后以9 J/cm2的剂量进行UVA照射30分钟。照射后，将HDFs与接种在Transwell上的HAMSCs共培养。共培养72小时后，进行衰老相关β-半乳糖苷酶染色，并通过流式细胞术分析检测HDFs中的活性氧（ROS）含量和线粒体膜电位（Δψm）。结果表明，与未共培养的组相比，与人HAMSCs共培养时，通过X-gal染色检测到的HDFs百分比显著降低。与单独用UVA处理的细胞相比，用UVA和HAMSCs处理的细胞中ROS含量降低，线粒体膜电位（Δψm）恢复。逆转录-定量聚合酶链反应揭示了HAMSCs对HDF衰老标记基因p53和基质金属蛋白酶-1 mRNA表达的显著影响。除此之外，蛋白质印迹分析证实了HAMSCs对UVA诱导衰老的作用，为新型再生治疗方法提供了基础。此外，结果表明细胞外信号调节激酶1/2丝裂原活化蛋白激酶信号转导途径的激活对于HAMSC介导的UVA保护作用至关重要。ROS含量的降低还表明HAMSCs未来可能具有治疗氧化应激介导的UVA皮肤衰老的潜力。
