Non-thermal Quantum Molecular Resonance electrotherapy and a blood-derived secretome protect oxidatively stressed human RPE cells by reprogramming redox, mitochondrial and immune–ECM networks

Background Age-related macular degeneration (AMD) is driven by chronic oxidative stress, parainflammation, and extracellular matrix (ECM) remodeling converging on the retinal pigment epithelium (RPE). Quantum molecular resonance (QMR) electrotherapy and patient blood-derived (PBD) secretomes represent non-pharmacologic approaches with cytoprotective and immunomodulatory potential; however, their transcriptional impact on oxidatively stressed RPE remains unknown. Methods ARPE-19 cells were exposed to tert-butyl hydroperoxide (tBHP) and treated with QMR, PBD secretome, or their combination (QMR+SECRETOME). Cell viability was assessed by MTT assay. Bulk RNA sequencing with Gene Ontology, KEGG, and Reactome enrichment—including time-resolved contrasts at 8, 24, and 72 h—profiled transcriptional responses across five groups (CTRL, CTRL-OX, QMR, SECRETOME, QMR+SECRETOME). Key differentially expressed genes were validated by RT-qPCR. Results QMR preserved ARPE-19 viability and induced multilayered transcriptional reprogramming encompassing upregulation of glutathione biosynthesis enzymes (GCLC, GCLM, GSR, GSS), NADPH-generating pathways (G6PD, IDH1/2), heme–iron-handling genes (HMOX1/2, FTH1), mitochondrial respiratory chain and mitophagy modules, and proteostasis networks (HSP70/90, ATG4 effectors, ubiquitin–proteasome components). The PBD secretome remodeled the RPE–microenvironment interface by engaging NF-κB cytokine/chemokine signaling (TNF, IL1B, IL6, CXCL8), ECM organization (collagens, MMPs, integrins), complement–coagulation cascades, lipid handling, and angiogenic pathways (VEGFA). Combined QMR+SECRETOME produced the largest transcriptional shift (2,240 DEGs), integrating both signatures into a convergent yet non-redundant stress-adaptation state. Temporal analyses revealed rapid QMR-mediated induction at 8 h, with progressive secretome-driven immune and ECM remodeling from 24 to 72 h. Conclusions QMR and PBD secretome engage complementary transcriptional programs in oxidatively stressed RPE: QMR strengthens intracellular antioxidant, mitochondrial, and proteostatic resilience, whereas the secretome reprograms immune and ECM signaling at the RPE–microenvironment interface. Their combination generates a cytoprotective state not achieved by either intervention alone, providing a mechanistic rationale for multimodal non-pharmacologic strategies in early AMD and RPE-centric degenerative diseases.