Wang Si-Wei, Li Ping, Liu Shi-Yu, Huang De-Lian, Zhang Si-Jia, Zeng Xi-Xi, Lan Tian, Mao Kai-Li, Gao Yuan, Cheng Yi-Fan, Shen Qing, Ruan Ye-Ping, Mao Zhu-Jun
School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou 310053, China; Panvascular Diseases Research Center, the Quzhou Affiliated Hospital of Wenzhou Medical University, Quzhou People's Hospital, Quzhou 324000, China.
School of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Phytomedicine. 2025 Mar;138:156408. doi: 10.1016/j.phymed.2025.156408. Epub 2025 Jan 18.
Resistance to senescence in retinal pigment epithelial (RPE) cells can delay the progression of age-related macular degeneration (AMD). However, the mechanisms underlying RPE cell senescence remain inadequately understood, and effective therapeutic strategies are lacking. While astragaloside IV (Ast) has demonstrated anti-aging properties, its specific effects on RPE cell senescence and potential mechanisms are not yet fully clarified.
This study aimed to explore the impacts of Ast on RPE cell senescence and to uncover the molecular mechanisms involved.
The therapeutic efficacy of Ast was assessed using sodium iodate (NaIO)-induced adult retinal pigment epithelial cell line 19 (ARPE-19) cell models and an AMD mouse model. To investigate the mechanisms by which Ast mitigated RPE cell senescence, RNA sequencing (RNA-seq), drug affinity responsive target stability-mass spectrometry (DARTS-MS), cellular thermal shift assay (CETSA), reverse transcription quantitative PCR (RT-qPCR), as well as western blotting were conducted.
Ast significantly inhibited NaIO-treated ARPE-19 cell senescence and protected against NaIO-induced AMD in mice. RNA-seq analysis revealed that Ast significantly attenuated inflammation-related signaling pathways and reduced the mRNA levels of interleukin-1 beta (IL-1β). Specifically, Ast decreased the stability of IL-1β mRNA while enhancing its N6-methyladenosine (mA) methylation. Furthermore, Ast directly interacted with fat mass and obesity-associated protein (FTO). Knockdown or pharmacological inhibition of FTO mitigated the senescence and IL-1β expression in NaIO-treated ARPE-19 cells. FTO was essential for Ast to inhibit cellular senescence and IL-1β expression. Additionally, inhibition or knockdown of FTO conferred also provided resistance to AMD in the murine model.
Our results indicated that Ast significantly attenuated RPE cell senescence and showed anti-AMD properties. FTO was demonstrated to be a promising therapeutic target for AMD treatment. These findings may provide a deeper understanding of the molecular mechanisms underlying RPE cell senescence in AMD and offer potential strategies for its prevention and management.
视网膜色素上皮（RPE）细胞对衰老的抗性可延缓年龄相关性黄斑变性（AMD）的进展。然而，RPE细胞衰老的潜在机制仍未得到充分了解，且缺乏有效的治疗策略。虽然黄芪甲苷（Ast）已显示出抗衰老特性，但其对RPE细胞衰老的具体作用及潜在机制尚未完全阐明。
本研究旨在探讨Ast对RPE细胞衰老的影响，并揭示其相关分子机制。
使用碘酸钠（NaIO）诱导的成年视网膜色素上皮细胞系19（ARPE-19）细胞模型和AMD小鼠模型评估Ast的治疗效果。为研究Ast减轻RPE细胞衰老的机制，进行了RNA测序（RNA-seq）、药物亲和力响应靶点稳定性-质谱分析（DARTS-MS）、细胞热迁移分析（CETSA）、逆转录定量PCR（RT-qPCR）以及蛋白质免疫印迹分析。
Ast显著抑制了NaIO处理的ARPE-19细胞衰老，并保护小鼠免受NaIO诱导的AMD影响。RNA-seq分析显示，Ast显著减弱炎症相关信号通路，并降低白细胞介素-1β（IL-1β）的mRNA水平。具体而言，Ast降低了IL-1β mRNA的稳定性，同时增强了其N6-甲基腺苷（m⁶A）甲基化。此外，Ast直接与脂肪量和肥胖相关蛋白（FTO）相互作用。敲低或药物抑制FTO可减轻NaIO处理的ARPE-19细胞的衰老和IL-1β表达。FTO对Ast抑制细胞衰老和IL-1β表达至关重要。此外，抑制或敲低FTO在小鼠模型中也赋予了对AMD的抗性。
我们的结果表明，Ast显著减轻RPE细胞衰老并显示出抗AMD特性。FTO被证明是AMD治疗的一个有前景的治疗靶点。这些发现可能为深入了解AMD中RPE细胞衰老的分子机制提供帮助，并为其预防和管理提供潜在策略。
