Department of Hematology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, China.
The First Clinical College of Zhejiang Chinese Medical University, Hangzhou, China.
Int Immunopharmacol. 2023 May;118:110131. doi: 10.1016/j.intimp.2023.110131. Epub 2023 Apr 5.
Panaxadiol saponin (PND) is a latent targeted drug for the treatment of aplastic anemia (AA). In this study, we examined the effects of PND on ferroptosis in iron-overload AA and Meg-01 cells. We utilized RNA-seq to analyze differentially expressed genes in iron-induced Meg-01 cells treated with PND. The effects of PND or combined with deferasirox (DFS) on iron deposition, labile iron pool (LIP), several ferroptosis events, apoptosis, mitochondrial structure, as well as ferroptosis-, Nrf2/HO-1-, and PI3K/AKT/mTOR pathway-related markers in iron-induced Meg-01 cells were examined by Prussian-blue staining, flow cytometer, ELISA, Hoechst 33342 staining, transmission electron microscope, and Western blot assays, respectively. Additionally, an AA mice model with iron overload was established. Then, the blood routine was assessed, and the number of bone marrow-derived mononuclear cells (BMMNCs) in mice was counted. Also, serum iron, ferroptosis events, apoptosis, histology, T lymphocyte percentage, ferroptosis-, Nrf2/HO-1-, and PI3K/AKT/mTOR signaling-related targets in primary megakaryocytes of AA mice with iron overload were assessed by commercial kits, TUNEL staining, hematoxylin and eosin (H&E) staining, Prussian blue staining, flow cytometer, and qRT-PCR analysis, respectively. PND suppressed iron-triggered iron overload, and apoptosis, and ameliorated mitochondrial morphology in Meg-01 cells. Importantly, PND ameliorated ferroptosis-, Nrf2/HO-1-, and PI3K/AKT/mTOR signaling-related marker expressions in iron-induced Meg-01 cells or primary megakaryocytes of AA mice with iron overload. Moreover, PND ameliorated body weight, peripheral blood cell counts, the number of BMMNCs, and histological injury in the iron-overload AA mice. Also, PND improved the percentage of T lymphocytes in the iron-overload AA mice. PND attenuates ferroptosis against iron-overload AA mice and Meg-01 cells via activating Nrf2/HO-1 and PI3K/AKT/mTOR pathway and is a promising novel therapeutic candidate for AA.
人参二醇皂苷（PND）是治疗再生障碍性贫血（AA）的潜在靶向药物。在这项研究中，我们研究了 PND 对铁过载 AA 和 Meg-01 细胞中铁死亡的影响。我们利用 RNA-seq 分析了 PND 处理铁诱导的 Meg-01 细胞中差异表达的基因。通过普鲁士蓝染色、流式细胞仪、ELISA、Hoechst 33342 染色、透射电子显微镜和 Western blot 检测 PND 或联合去铁酮（DFS）对铁诱导的 Meg-01 细胞中铁沉积、不稳定铁池（LIP）、几种铁死亡事件、细胞凋亡、线粒体结构以及铁死亡、Nrf2/HO-1 和 PI3K/AKT/mTOR 通路相关标志物的影响。此外，建立了铁过载 AA 小鼠模型。然后，评估血液常规，计数小鼠骨髓来源的单核细胞（BMMNC）的数量。还通过商业试剂盒、TUNEL 染色、苏木精和伊红（H&E）染色、普鲁士蓝染色、流式细胞仪和 qRT-PCR 分析评估血清铁、铁死亡事件、细胞凋亡、组织学、AA 小鼠铁过载原代巨核细胞中 T 淋巴细胞百分比、铁死亡、Nrf2/HO-1 和 PI3K/AKT/mTOR 信号相关靶点。PND 抑制铁触发的铁过载、细胞凋亡并改善 Meg-01 细胞中线粒体形态。重要的是，PND 改善了铁诱导的 Meg-01 细胞或 AA 小鼠铁过载原代巨核细胞中铁死亡、Nrf2/HO-1 和 PI3K/AKT/mTOR 信号相关标志物的表达。此外，PND 改善了铁过载 AA 小鼠的体重、外周血细胞计数、BMMNC 数量和组织学损伤。PND 还改善了铁过载 AA 小鼠中 T 淋巴细胞的百分比。PND 通过激活 Nrf2/HO-1 和 PI3K/AKT/mTOR 通路减轻铁过载 AA 小鼠和 Meg-01 细胞中的铁死亡，是一种有前途的 AA 新型治疗候选药物。
