Department of Plant Physiology, Faculty of Science, Trakya University, 22030, Edirne, Turkey.
Department of Medical Biology, Faculty of Medicine, Trakya University, 22030, Edirne, Turkey.
Environ Sci Pollut Res Int. 2022 Mar;29(12):17977-17996. doi: 10.1007/s11356-021-17039-8. Epub 2021 Oct 22.
Increasing evidence supports the existence of cross-kingdom gene regulation. However, the therapeutic potential of stress-specific plant miRNAs and their role in UV-related pathologies in human tissue remain largely unexplored. The aim of this study was to investigate the therapeutic potential and mechanisms of action of stress-induced miRNA cocktails (SI-WmiRs) from Einkorn wheat (Triticum monococcum L.) on human keratinocyte (HaCaT) cells exposed to a high dose of UV-B radiation. We used a biofactory approach and irradiated wheatgrass with UV-C for 240 min to obtain the specific SI-WmiRs that wheat produces to recover from UV stress. We followed the plant with molecular and biochemical analyses and extracted our SI-WmiRs at the most appropriate time (0 h and 6 h after UV-C application). Then, we applied the SI-WmiR cocktail to HaCaT cells exposed to high-dose of UV-B radiation. Our results show that UV-B radiation induced lipid peroxidation and DNA damage, as demonstrated by increased malondialdehyde (MDA) levels and changes in the RAPD band profile, respectively. UV stress also impaired IL6/JAK2/STAT3 signalling and activated the inflammatory mediators IL6 and TNF-α in HaCaT cells, leading to significant induction of apoptotic cell death. We found that SI-WmiR transfection prevents lipid peroxidation and oxidative stress-related DNA damage by increasing antioxidant (CuZn-SOD, Mn-SOD) and DNA repair (EXO1, SMUG1 and XRCC3) gene expression. In addition, SI-WmiRs regulated IL6/JAK2/STAT3 signalling by reducing JAK2 and STAT3 gene expression and phosphorylated protein levels compared to the control treatments. Moreover, SI-WmiRs inhibited pro-apoptotic BAX, Caspase 3 and Caspase 8 gene expression and protein levels to prevent apoptosis of UV-stressed HaCaT cells. Our results demonstrate that stress-induced wheat miRNAs produced using a biofactory approach have strong potential as a novel and effective alternative therapy for UV stress-related skin damage.
越来越多的证据支持跨王国基因调控的存在。然而，应激特异性植物 miRNA 的治疗潜力及其在人类组织中与 UV 相关的病理中的作用在很大程度上仍未得到探索。本研究旨在研究源自单粒小麦（Triticum monococcum L.）的应激诱导 miRNA 鸡尾酒（SI-WmiRs）对暴露于高剂量 UV-B 辐射的人角质形成细胞（HaCaT）的治疗潜力和作用机制。我们使用生物工厂方法用 UV-C 辐照小麦草 240 分钟，以获得小麦产生的特定 SI-WmiRs，以从 UV 应激中恢复。我们通过分子和生化分析对植物进行跟踪，并在最合适的时间（UV-C 应用后 0 小时和 6 小时）提取我们的 SI-WmiRs。然后，我们将 SI-WmiR 鸡尾酒应用于暴露于高剂量 UV-B 辐射的 HaCaT 细胞。我们的结果表明，UV-B 辐射诱导脂质过氧化和 DNA 损伤，这分别表现为丙二醛（MDA）水平升高和 RAPD 带谱变化。UV 应激还损害了 IL6/JAK2/STAT3 信号通路，并激活了 HaCaT 细胞中的炎症介质 IL6 和 TNF-α，导致凋亡细胞死亡的显著诱导。我们发现，通过增加抗氧化（CuZn-SOD、Mn-SOD）和 DNA 修复（EXO1、SMUG1 和 XRCC3）基因的表达，SI-WmiR 转染可防止脂质过氧化和与氧化应激相关的 DNA 损伤。此外，与对照处理相比，SI-WmiRs 通过降低 JAK2 和 STAT3 基因表达和磷酸化蛋白水平来调节 IL6/JAK2/STAT3 信号通路。此外，SI-WmiRs 抑制促凋亡 BAX、Caspase 3 和 Caspase 8 基因表达和蛋白水平，以防止 UV 应激的 HaCaT 细胞凋亡。我们的结果表明，使用生物工厂方法产生的应激诱导的小麦 miRNA 具有作为治疗与 UV 应激相关的皮肤损伤的新型有效替代疗法的巨大潜力。
