ABSTRACT
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.
Subject(s)
Anemia, Aplastic , Ferroptosis , Iron Overload , Saponins , Mice , Animals , Proto-Oncogene Proteins c-akt/metabolism , NF-E2-Related Factor 2/genetics , NF-E2-Related Factor 2/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Saponins/pharmacology , Saponins/therapeutic use , Anemia, Aplastic/drug therapy , Signal Transduction , Iron Overload/drug therapy , TOR Serine-Threonine Kinases/metabolism , IronABSTRACT
Objective To explore the effect of panaxadiols saponin (PDS) on the oxidative damage in lung tissue in hemorrhage-lipopolysaccharides (LPS) two hits rats. Methods Forty Wistar rats were divided randomly into 4 groups: sham operational group(S group); two hits groups with hemorrhage-LPS(HL group);dexamethasone preventive therapy group (HLD group) ;panaxadiol saponins preventive therapy group(HLP group). The rats were made first-hit by hemorrhagic shock,LPS were administered intraperitoneally to make endotoxic shock and induce two hits in rats,then the model of acute lung injury (ALI) induced by two hits had been built up successfully. NO-2/NO-3,MDA content and NOS,iNOS,SOD activities were measured by 722S spectrophotometer. Results The results of pathological observation showed that there was obvious inflammatory reaction in lung tissues after two-hits,and the structure was destroied.Compared with HL group,the inflammatory reaction was reduced in HLD group and HLD group.NOS,iNOS activities and NO-2/NO-3,MDA contents were increased significantly,and SOD activity was significantly lower in HL group compared with S group(P
